CBD Oil For Sebaceous Cysts

CBD Oil For Sebaceous Cysts “I have a cyst in my jaw.” Hard to say. I have used CBD for other types of cysts with good success, in particular the skin, spinal cord, and liver. Standard doses Finding a cystic acne treatment that actually works is hard. This cystic acne spot treatment cream utilizes a two staged approach to skin rejuvination. Cannabidiol exerts sebostatic and antiinflammatory effects on human sebocytes 1 DE-MTA “Lendület” Cellular Physiology Research Group, Department of Physiology, University of Debrecen, Debrecen,

CBD Oil For Sebaceous Cysts

“I have a cyst in my jaw.”

Hard to say. I have used CBD for other types of cysts with good success, in particular the skin, spinal cord, and liver. Standard doses of 30 mg have been effective in most cases. Topical applications for accessible cysts can also be effective. I apply CBD concentrates to all wounds, burns, and viral blisters finding immediate pain relief and enhanced healing.

I know this is a bit late but for posterity, I have an arachnoidal cyst and subsequent hygroma. I am a doctor, but not that kind. The pain and seizures brought on at first were crippling. I utilize cannabis both recreationally and medically. The choice in my mind became, either let them crack open my skull or beat it myself. I have chosen the later. I vaporize the CBD, it’s a bit more expensive but easier for me. I refuse to take opiates so my pain relief options were slim. After a ‘strict’ regiment of the CBD the headaches have all but gone. I’m going to go in for a contrast MRI here in a month or so and comapre it to the ones taken back in October. My bet, the thing shrunk. Again, I’m not a medical doctor but I am a scientist.
Best of luck.
Peace and Namaste
Rev. Michael Aaron Knight Ph.D

How to Get Rid of Cystic Acne With CBD?

Curious about cystic acne treatment techniques that actually work? What about using CBD for sebaceous cyst relief? Do cystic acne spot treatment creams exit that might help?

Well, you’re going to love this article!

Have you ever experienced going to sleep with a nice, clear face and then getting up with swollen zits on your face? Not the best surprise, is it? This is annoying, to say the least!

The first thing that comes to mind is usually, “Oh my God, how can I get rid of this as quickly as possible?”

How about having an important business engagement, or date. No one wants to have all eyes on them when their skin looks, well, bad. While you might be able to rub some cystic acne cream on your face and call it good, studies suggest that most cystic acne cream doesn’t work that well.

Don’t worry, though. This article contains all you need to know about how a cystic acne spot treatment might help, and cystic acne treatment options with the use of a special product that uses CBD and salicylic acid.

Cystic Acne: A Horrible Experience for All

Once zits appear, it usually takes more time than we want for them to go away. The Mayo Clinic has declared cystic acne as a more serious form of acne that sprouts its roots deep below the skin.

Cystic acne can be super sore and results in significant, red, swelling and constant pain that surrounds the area.

What causes cystic acne is important to know. The main reasons for cystic acne development are:

  • excessive sebum secretion
  • production of bacteria into the pores
  • dead skin cells clogging the pores

Cystic acne, to some extent, looks like boils and is typically extremely painful if touched. Furthermore, this type of acne usually occurs on the face, neck, shoulders, back, and arms. It is usually the form of acne that leaves little divots in the skin than result in severe scarring.

A good cystic acne spot treatment will clear the acne before it is able to leave these divots.

Whatever product you end up using, just remember the cystic acne spot treatment must be extremely effective at both clearing the pore, and healing the affected area.

What good is a spot treatment that clears up an area, but leaves it red and swollen for days? We’re all looking to get rid of that redness as soon as humanly possible, am I right?

Example of cystic acne

Cystic acne is more common in people with oily skin and is usually found in teens, women, and older adults with hormonal imbalances.

If you think you may be suffering from cystic acne, you need to consult a dermatologist for the best possible advice in regards to both the cause of your issues, and the best things to use as a cystic acne spot treatment.

New Phase Blends has a product called ‘glow’ that can treat skin issues like this, but nothing compares to having a one-on-one consult with a dermatologist. Prescription medications may be necessary.

At New Phase Blends, we create products to help people not need to rely on prescription drugs, however, there is nothing wrong with taken them as needed. Some people require them, and some do not – there is nothing wrong with that!

For your best defense against cystic acne or sebaceous cysts, we have compiled a list of a few home remedies to help clear your skin up as quickly and safely as possible. These remedies can ease the condition of the skin and diminish the severity of your skin troubles.

So, let’s get started!

How To Identify Cystic Acne or Sebaceous Cysts

It is very important to identify the type of acne you may have in order to best develop a plan to clear your skin.

Cystic Acne

Cystic acne is the most severe form of acne that can result in extreme pain and redness. Unlike other acne types that remain on the top of the skin surface, cystic acne resembles the boils, and roots deeper below the skin.

Since this type of acne can be quite severe, a powerful cystic acne spot treatment is normally required.

Redness, large pus-filled cysts, large white bumps, or being painful to the touch are few other identifiable cystic acne symptoms.

The area on our bodies that are more likely to develop cystic acne include the face, chest, neck, back, arms, shoulders, and even armpits.

At the same time, the face is the most noticeable area with cystic acne. Most people can manage this by applying a cystic acne spot treatment on their back and shoulders. The spot treatments can also be applied to the face, but you can’t really cover up your face and hide the acne like you can for your back or shoulders.

For this reason, we all want something that works fast, and well.

Sebaceous Cysts

Cysts, generally speaking, are simply spots in the body (typically swollen in appearance) that may have a liquid or gel-like substance inside of them. Sebaceous cysts are more likely to be found on the skin of the neck, face, or upper body.

hey aren’t necessarily dangerous, and they grow quite slow when compared to other forms of cysts.

While not life-threatening, you should be sure to have them checked on periodically. Some people prefer to have them removed, some people prefer to leave them alone – it all depends on the size, location, and severity of the sebaceous cyst.

Photo Source: WebMD

As you can see in the picture above, sebaceous cysts can be large, but they aren’t accompanied by the unfortunate redness that covers the area of cystic acne.

Sebaceous cysts will sometimes require a different approach than a cystic acne spot treatment can offer. Dermatologists may need to remove the cyst themselves.

Have you heard of Dr. Pimple Popper? She actually built a booming business on airing some of her sebaceous cyst removals.

Cystic Acne Treatment: Over the Counter Creams

The following home-based remedies are generally safe to treat cystic acne, however, before trying these remedies, consult with your dermatologist.

They will offer you medical advice that we cannot otherwise offer on this website. Also, please remember that CBD products are not approved by the FDA as a dietary supplement, yet.

The statements or products contained on this website are not meant to diagnose, treat, prevent, or cure any form of disease or skin conditions.

Now, let’s find out how to get rid of cystic acne, or at least minimize the appearance of it.

CBD Oil & CBD Products for Cystic Acne Treatment

Cannabidiol, also known as CBD, is getting hype and becoming famous for its pain-relieving and anti-inflammatory properties. CBD has been found as the best natural source for treating acne and other skin problems more naturally.

However, CBD alone isn’t the best approach to treating cystic acne or sebaceous cysts. While CBD may, in fact, aid in reducing inflammation and swelling that is often associated with breakouts, it cannot clean your pores out.

New Phase Blends developed a product called ‘glow’ that utilizes a 1% salicylic acid base, in addition to tea tree oil and CBD-rich hemp extract. All of these together help trigger a powerful two-staged acne fighting product when applied directly to the skin.

While the salicylic acid beings to clear out the pores and surrounding area, the CBD-rich hemp extract reduces redness and swelling. Remember, the worst part about most cystic acne spot treatments is they don’t address the redness and swelling.

We’ve fixed that problem.

You can apply the CBD products topically as a cystic acne treatment to get some relief from the pain and swelling.

Plus, CBD topicals such as creams, balms, and lotions are readily available for direct skin application. CBD drops, or CBD tinctures, are also a good product to use to help take the edge off the pain and swelling associated with cystic acne or other forms of skin issues.

Cystic acne cream, like ‘glow’, can be a great approach to helping get your skin looking nice and healthy again.

Ice Cubes

Ice has soothing effects for swollen and itchy skin.

It helps in decreasing swelling, pain, itchiness, and redness on any area of the skin. Ice is often useful in diminishing the severity of cystic acne. Rubbing the ice cube almost three times a day on cystic acne decreases the swelling and relieves the pain.

Ice cubes, while soothing, are only a very temporary approach to cystic acne treatment, and are perhaps the weakest form of a cystic acne spot treatment.

Aspirin Mask

An aspirin mask can help to reduce the inflammation and pain caused by cystic acne. A few home remedies have suggested using the mixture of crushed aspirin powder and water on cystic acne. The application of this mild, thick paste acts as a type of cystic acne spot treatment.

What, exactly, is an aspirin mask? It’s a thick, pasty-type substance that is made with crushed up aspirin and small amounts of luke-warm water. The water should be warm, at a minimum, to promote opening of the pores. This allows the aspirin mask to penetrate your skin as deeply as possible.

The company Popsugar has a pretty good article on making the best aspirin mask. Check it out here.

This remedy is not for every skin type because few people have a known allergy to salicylates (contained in aspirin products).

Direct application of aspirin on the skin can irritate it so, please double check with your doctor or dermatologist before trying this.

Balanced Diet

People in favor of natural remedies for cystic acne treatment have suggested keeping a strict check on their diet routine. Dairy and dairy-based items can help develop cystic acne (in some people).

To treat the cystic acne, you may need to avoid dairy items, including milk, yogurt, cheese, and butter, for at least three to four weeks to give your skin a bit of relief.

If there is no new cystic acne eruption, that means dairy could have very well been the cause of your skin trouble.

Also, excessive use of sugar, oil, and processed food can trigger cystic acne breakouts. Focus on a healthy diet plan rich in fiber, protein, and vitamins/minerals.

A great cystic acne treatment will encompass many different things, including a balanced diet.

Turmeric Mask

This readily available kitchen item, turmeric, is a natural solution to many skin problems, including getting rid of cystic acne. It is full of anti-inflammatory and antiseptic properties that can help decrease the severity of cystic acne.

Apply small amounts of turmeric powder mixed with warm water and directly apply it to the cystic acne. Let it penetrate the skin for 45 minutes or so and then rinse it off. Apply this paste two times a day for better results.

Take care while applying it directly on the skin as it can irritate sensitive skin. Just like with the aspirin mask, make sure the water is warm to help open the pores. This lets the pores absorb even more of the turmeric.

Vinegar Cleanser

Apple cider vinegar is an excellent natural remedy to get rid of cystic acne. It has anti-bacterial properties that help to remove the bacteria and dead skin cells from the skin.

Some proponents of natural healing remedies have suggested using a diluted white vinegar cleanser two times a day.

Be careful when applying a vinegar and water solution, as it can irritate your skin. Also, avoid the direct application of vinegar on the face. A good ratio of vinegar to water is two capfuls of vinegar and three cups of filtered, clean water.

Probiotics

The daily dose of probiotics encourages both skin health and gut health. Probiotics decrease skin inflammation and can lead to a much more clear and healthy skin. A good approach to cystic acne treatment might involve the use of these healthy probiotics.

Add a great source of probiotics such as yogurt, kimchi, vegetables, kefir, and other foods containing beneficial bacteria and help regain skin health. There are also probiotics available in pill form.

While probiotics aren’t a good cystic acne spot treatment, since they must be digested in the gut, they do help clear up your skin indirectly.

Tea Tree Oil

Tea tree oil is a a natural solution to soothing the skin and also promotes skin health. It contains antimicrobial and anti-inflammatory properties that make it an excellent option for treating and preventing cystic acne.

As with any other oil, be careful with potentially getting your skin too oily. Some people are ok with oil-based facial products, some are not. Everyone’s skin is a little bit different and reacts to different products…differently.

What Are The Medical Cystic Acne Treatment Options?

Home remedies for cystic acne treatment or sebaceous cysts should be the first choice for safe treatment, however, it tends to be time-consuming.

These choices may not be potent enough to treat cystic acne. Remember when I mentioned earlier that some people may end up needing a prescription approach for their skin problems? Some of the medical treatments for severe cystic acne or sebaceous cysts may include:

  • azelaic acid (Azelex)
  • isotretinoin (Accutane)
  • oral antibiotics (tetracycline)
  • dapsone (Aczone)
  • spironolactone (Aldactone)
  • topical retinoids (Retin-A)
  • Light-based therapy (laser or photodynamic)
  • steroid injections into cystic and nodular wounds.

*Consult with your dermatologist before undergoing any medical treatments.

Conclusion | Cystic Acne Treatment

Undoubtedly, natural healing options are a good first step in a good cystic acne treatment regimen.

This can sometimes be done with easy methods as mentioned above. Other times, it requires the use of a cystic acne spot treatment, similar to glow.

Remember, the most important factor in cystic acne treatment is finding a product that not only cleans and clears out your skin, but corrects the redness and swelling too. Many products on the market can clean the skin, but they leave it dry and red.

See also  Vapor Mint With CBD Oil

We don’t want this.

Give our cystic acne cream products a try, and don’t be scared or ashamed to consult a dermatologist for extra help if needed. That’s what they are there for!

Finally, if you aren’t pleased with our products (any of them) please return them within 30 days of the original purchase for your money back – all of it. We want you to love our products – nothing less.

Cannabidiol exerts sebostatic and antiinflammatory effects on human sebocytes

1 DE-MTA “Lendület” Cellular Physiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 2 Laboratory for Ion Channel Research and TRP Research Platform Leuven, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium. 3 Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan. 4 Department of Dermatology, University of Lübeck, Lübeck, Germany. 5 Neurobiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 6 Laboratory of Cutaneous Physiopathology and Integrated Center of Metabolomics Research, San Gallicano Dermatologic Institute, IRCCS, Rome, Italy. 7 Departments of Dermatology, Venereology, and Allergology and Immunology, Dessau Medical Center, Dessau, Germany. 8 School of Translational Medicine, University of Manchester, Manchester, United Kingdom.

Balázs I. Tóth

1 DE-MTA “Lendület” Cellular Physiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 2 Laboratory for Ion Channel Research and TRP Research Platform Leuven, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium. 3 Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan. 4 Department of Dermatology, University of Lübeck, Lübeck, Germany. 5 Neurobiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 6 Laboratory of Cutaneous Physiopathology and Integrated Center of Metabolomics Research, San Gallicano Dermatologic Institute, IRCCS, Rome, Italy. 7 Departments of Dermatology, Venereology, and Allergology and Immunology, Dessau Medical Center, Dessau, Germany. 8 School of Translational Medicine, University of Manchester, Manchester, United Kingdom.

István Borbíró

1 DE-MTA “Lendület” Cellular Physiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 2 Laboratory for Ion Channel Research and TRP Research Platform Leuven, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium. 3 Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan. 4 Department of Dermatology, University of Lübeck, Lübeck, Germany. 5 Neurobiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 6 Laboratory of Cutaneous Physiopathology and Integrated Center of Metabolomics Research, San Gallicano Dermatologic Institute, IRCCS, Rome, Italy. 7 Departments of Dermatology, Venereology, and Allergology and Immunology, Dessau Medical Center, Dessau, Germany. 8 School of Translational Medicine, University of Manchester, Manchester, United Kingdom.

Koji Sugawara

1 DE-MTA “Lendület” Cellular Physiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 2 Laboratory for Ion Channel Research and TRP Research Platform Leuven, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium. 3 Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan. 4 Department of Dermatology, University of Lübeck, Lübeck, Germany. 5 Neurobiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 6 Laboratory of Cutaneous Physiopathology and Integrated Center of Metabolomics Research, San Gallicano Dermatologic Institute, IRCCS, Rome, Italy. 7 Departments of Dermatology, Venereology, and Allergology and Immunology, Dessau Medical Center, Dessau, Germany. 8 School of Translational Medicine, University of Manchester, Manchester, United Kingdom.

Attila G. Szöllõsi

1 DE-MTA “Lendület” Cellular Physiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 2 Laboratory for Ion Channel Research and TRP Research Platform Leuven, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium. 3 Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan. 4 Department of Dermatology, University of Lübeck, Lübeck, Germany. 5 Neurobiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 6 Laboratory of Cutaneous Physiopathology and Integrated Center of Metabolomics Research, San Gallicano Dermatologic Institute, IRCCS, Rome, Italy. 7 Departments of Dermatology, Venereology, and Allergology and Immunology, Dessau Medical Center, Dessau, Germany. 8 School of Translational Medicine, University of Manchester, Manchester, United Kingdom.

Gabriella Czifra

1 DE-MTA “Lendület” Cellular Physiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 2 Laboratory for Ion Channel Research and TRP Research Platform Leuven, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium. 3 Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan. 4 Department of Dermatology, University of Lübeck, Lübeck, Germany. 5 Neurobiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 6 Laboratory of Cutaneous Physiopathology and Integrated Center of Metabolomics Research, San Gallicano Dermatologic Institute, IRCCS, Rome, Italy. 7 Departments of Dermatology, Venereology, and Allergology and Immunology, Dessau Medical Center, Dessau, Germany. 8 School of Translational Medicine, University of Manchester, Manchester, United Kingdom.

Balázs Pál

1 DE-MTA “Lendület” Cellular Physiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 2 Laboratory for Ion Channel Research and TRP Research Platform Leuven, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium. 3 Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan. 4 Department of Dermatology, University of Lübeck, Lübeck, Germany. 5 Neurobiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 6 Laboratory of Cutaneous Physiopathology and Integrated Center of Metabolomics Research, San Gallicano Dermatologic Institute, IRCCS, Rome, Italy. 7 Departments of Dermatology, Venereology, and Allergology and Immunology, Dessau Medical Center, Dessau, Germany. 8 School of Translational Medicine, University of Manchester, Manchester, United Kingdom.

Lídia Ambrus

1 DE-MTA “Lendület” Cellular Physiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 2 Laboratory for Ion Channel Research and TRP Research Platform Leuven, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium. 3 Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan. 4 Department of Dermatology, University of Lübeck, Lübeck, Germany. 5 Neurobiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 6 Laboratory of Cutaneous Physiopathology and Integrated Center of Metabolomics Research, San Gallicano Dermatologic Institute, IRCCS, Rome, Italy. 7 Departments of Dermatology, Venereology, and Allergology and Immunology, Dessau Medical Center, Dessau, Germany. 8 School of Translational Medicine, University of Manchester, Manchester, United Kingdom.

Jennifer Kloepper

1 DE-MTA “Lendület” Cellular Physiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 2 Laboratory for Ion Channel Research and TRP Research Platform Leuven, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium. 3 Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan. 4 Department of Dermatology, University of Lübeck, Lübeck, Germany. 5 Neurobiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 6 Laboratory of Cutaneous Physiopathology and Integrated Center of Metabolomics Research, San Gallicano Dermatologic Institute, IRCCS, Rome, Italy. 7 Departments of Dermatology, Venereology, and Allergology and Immunology, Dessau Medical Center, Dessau, Germany. 8 School of Translational Medicine, University of Manchester, Manchester, United Kingdom.

Emanuela Camera

1 DE-MTA “Lendület” Cellular Physiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 2 Laboratory for Ion Channel Research and TRP Research Platform Leuven, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium. 3 Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan. 4 Department of Dermatology, University of Lübeck, Lübeck, Germany. 5 Neurobiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 6 Laboratory of Cutaneous Physiopathology and Integrated Center of Metabolomics Research, San Gallicano Dermatologic Institute, IRCCS, Rome, Italy. 7 Departments of Dermatology, Venereology, and Allergology and Immunology, Dessau Medical Center, Dessau, Germany. 8 School of Translational Medicine, University of Manchester, Manchester, United Kingdom.

Matteo Ludovici

1 DE-MTA “Lendület” Cellular Physiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 2 Laboratory for Ion Channel Research and TRP Research Platform Leuven, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium. 3 Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan. 4 Department of Dermatology, University of Lübeck, Lübeck, Germany. 5 Neurobiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 6 Laboratory of Cutaneous Physiopathology and Integrated Center of Metabolomics Research, San Gallicano Dermatologic Institute, IRCCS, Rome, Italy. 7 Departments of Dermatology, Venereology, and Allergology and Immunology, Dessau Medical Center, Dessau, Germany. 8 School of Translational Medicine, University of Manchester, Manchester, United Kingdom.

Mauro Picardo

1 DE-MTA “Lendület” Cellular Physiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 2 Laboratory for Ion Channel Research and TRP Research Platform Leuven, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium. 3 Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan. 4 Department of Dermatology, University of Lübeck, Lübeck, Germany. 5 Neurobiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 6 Laboratory of Cutaneous Physiopathology and Integrated Center of Metabolomics Research, San Gallicano Dermatologic Institute, IRCCS, Rome, Italy. 7 Departments of Dermatology, Venereology, and Allergology and Immunology, Dessau Medical Center, Dessau, Germany. 8 School of Translational Medicine, University of Manchester, Manchester, United Kingdom.

Thomas Voets

1 DE-MTA “Lendület” Cellular Physiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 2 Laboratory for Ion Channel Research and TRP Research Platform Leuven, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium. 3 Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan. 4 Department of Dermatology, University of Lübeck, Lübeck, Germany. 5 Neurobiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 6 Laboratory of Cutaneous Physiopathology and Integrated Center of Metabolomics Research, San Gallicano Dermatologic Institute, IRCCS, Rome, Italy. 7 Departments of Dermatology, Venereology, and Allergology and Immunology, Dessau Medical Center, Dessau, Germany. 8 School of Translational Medicine, University of Manchester, Manchester, United Kingdom.

Christos C. Zouboulis

1 DE-MTA “Lendület” Cellular Physiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 2 Laboratory for Ion Channel Research and TRP Research Platform Leuven, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium. 3 Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan. 4 Department of Dermatology, University of Lübeck, Lübeck, Germany. 5 Neurobiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 6 Laboratory of Cutaneous Physiopathology and Integrated Center of Metabolomics Research, San Gallicano Dermatologic Institute, IRCCS, Rome, Italy. 7 Departments of Dermatology, Venereology, and Allergology and Immunology, Dessau Medical Center, Dessau, Germany. 8 School of Translational Medicine, University of Manchester, Manchester, United Kingdom.

Ralf Paus

1 DE-MTA “Lendület” Cellular Physiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 2 Laboratory for Ion Channel Research and TRP Research Platform Leuven, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium. 3 Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan. 4 Department of Dermatology, University of Lübeck, Lübeck, Germany. 5 Neurobiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 6 Laboratory of Cutaneous Physiopathology and Integrated Center of Metabolomics Research, San Gallicano Dermatologic Institute, IRCCS, Rome, Italy. 7 Departments of Dermatology, Venereology, and Allergology and Immunology, Dessau Medical Center, Dessau, Germany. 8 School of Translational Medicine, University of Manchester, Manchester, United Kingdom.

Tamás Bíró

1 DE-MTA “Lendület” Cellular Physiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 2 Laboratory for Ion Channel Research and TRP Research Platform Leuven, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium. 3 Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan. 4 Department of Dermatology, University of Lübeck, Lübeck, Germany. 5 Neurobiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 6 Laboratory of Cutaneous Physiopathology and Integrated Center of Metabolomics Research, San Gallicano Dermatologic Institute, IRCCS, Rome, Italy. 7 Departments of Dermatology, Venereology, and Allergology and Immunology, Dessau Medical Center, Dessau, Germany. 8 School of Translational Medicine, University of Manchester, Manchester, United Kingdom.

1 DE-MTA “Lendület” Cellular Physiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 2 Laboratory for Ion Channel Research and TRP Research Platform Leuven, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium. 3 Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan. 4 Department of Dermatology, University of Lübeck, Lübeck, Germany. 5 Neurobiology Research Group, Department of Physiology, University of Debrecen, Debrecen, Hungary. 6 Laboratory of Cutaneous Physiopathology and Integrated Center of Metabolomics Research, San Gallicano Dermatologic Institute, IRCCS, Rome, Italy. 7 Departments of Dermatology, Venereology, and Allergology and Immunology, Dessau Medical Center, Dessau, Germany. 8 School of Translational Medicine, University of Manchester, Manchester, United Kingdom.

Address correspondence to: Tamás Bíró, University of Debrecen, Nagyerdei krt. 98. Debrecen, H-4032, Hungary. Phone: 3652.255.575; E-mail: [email protected]

Associated Data

Abstract

The endocannabinoid system (ECS) regulates multiple physiological processes, including cutaneous cell growth and differentiation. Here, we explored the effects of the major nonpsychotropic phytocannabinoid of Cannabis sativa, (-)-cannabidiol (CBD), on human sebaceous gland function and determined that CBD behaves as a highly effective sebostatic agent. Administration of CBD to cultured human sebocytes and human skin organ culture inhibited the lipogenic actions of various compounds, including arachidonic acid and a combination of linoleic acid and testosterone, and suppressed sebocyte proliferation via the activation of transient receptor potential vanilloid-4 (TRPV4) ion channels. Activation of TRPV4 interfered with the prolipogenic ERK1/2 MAPK pathway and resulted in the downregulation of nuclear receptor interacting protein-1 (NRIP1), which influences glucose and lipid metabolism, thereby inhibiting sebocyte lipogenesis. CBD also exerted complex antiinflammatory actions that were coupled to A2a adenosine receptor-dependent upregulation of tribbles homolog 3 (TRIB3) and inhibition of the NF-κB signaling. Collectively, our findings suggest that, due to the combined lipostatic, antiproliferative, and antiinflammatory effects, CBD has potential as a promising therapeutic agent for the treatment of acne vulgaris.

Introduction

Acne vulgaris is the most common human skin disease, affecting quality of life of millions worldwide. In spite of heroic basic and applied research efforts, we still lack indisputably curative anti-acne agents, which target multiple pathogenetic steps of acne (sebum overproduction, unwanted sebocyte proliferation, inflammation) and, moreover, which possess favorable side effect profiles (1, 2). Investigations over the last two decades have confirmed unambiguously that the human body expresses such receptors, which are able to specifically bind and recognize characteristic terpene-phenol compounds of the infamous plant Cannabis sativa, collectively referred to as phytocannabinoids. These receptors, their endogenous ligands (the endocannabinoids [eCBs]), and the enzymes involved in the synthesis and degradation of the eCBs collectively constitute the eCB system (ECS), a complex intercellular signaling network markedly involved in the regulation of various physiological processes (3–6).

Investigation of the cutaneous cannabinoid system seems to be a promising choice when searching for novel therapeutic possibilities (7, 8). Indeed, we have shown previously that the skin ECS regulates cutaneous cell growth and differentiation (9, 10), and it reportedly exerts antiinflammatory effects (11). Of further importance, we have also demonstrated that the ECS plays a key role in the regulation of sebum production (12). According to our recent findings, prototypic eCBs, such as N-arachidonoyl ethanolamide (anandamide [AEA]) and 2-arachidonoylglycerol, are constitutively produced in human sebaceous glands. Moreover, using human immortalized SZ95 sebocytes, we have also demonstrated that these locally produced eCBs (acting through a CB2 cannabinoid receptor→ERK1/2 MAPK→PPAR pathway) induce terminal differentiation of these cells, which is characterized by increased neutral lipid (sebum) production of the sebocytes (12). These findings confirmed unambiguously that human sebocytes have a functionally active ECS; yet, we did not possess data on the potential effect(s) of plant-derived cannabinoids.

(-)-Cannabidiol (CBD) is the most studied nonpsychotropic phytocannabinoid (13–15). It has already been applied in clinical practice without any significant side effects (Sativex) (16), and numerous ongoing phase II and III trials intend to explore its further therapeutic potential (17). Hence, within the confines of the current study, we intended to reveal the biological actions of CBD on the human sebaceous gland. Since we lack adequate animal models (18), we used human immortalized SZ95 sebocytes, the best available cellular system (19), and the full-thickness human skin organ culture (hSOC) technique (20).

Results

CBD normalizes “pro-acne agent”–induced excessive lipid synthesis of human sebocytes.

We first assessed the biological effects of CBD (1–10 μM) on the lipogenesis of SZ95 sebocytes. Although eCBs are known to show intense lipogenic actions via the metabotropic CB2 receptors (12), neither semiquantitative Oil Red O nor quantitative Nile Red staining indicated changes in the basal neutral (sebaceous) lipid synthesis of SZ95 sebocytes following 24-hour CBD treatment (Figure ​ (Figure1, 1 , A–C) (or 48-hour CBD treatment; data not shown). Intriguingly, however, CBD markedly inhibited the lipogenic action of the prototypic eCB, AEA, in a dose-dependent manner (1–10 μM; Figure ​ Figure1, 1 , C–E).

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We also tested its effect on actions of other lipogenic substances, which were shown previously to act through different, ECS-independent signal transduction mechanisms. Indeed, CBD effectively inhibited lipid synthesis induced by either arachidonic acid (AA) (21) or the combination of linoleic acid and testosterone (LA-T) (ref. 22 and Figure ​ Figure1F), 1 F), indicating that the effect of CBD is not “ECS specific” but a “universal” lipostatic action.

Since cannabinoids have been very often shown to exert “biphasic” effects (i.e., opposing physiological actions at nM vs. μM concentrations) (23), we also tested the effects of lower (1–100 nM) CBD concentrations; notably, they did not influence either basal or AA-induced lipid synthesis of the sebocytes (Supplemental Figure 1; supplemental material available online with this article; doi:10.1172/JCI64628DS1).

We also investigated the effects of CBD on the lipidome of SZ95 sebocytes under in vitro conditions that mimicked “acne-like” circumstances (the latter was achieved by using a key “pro-acne” inflammatory mediator, AA) (1, 2, 21, 24–26). Importantly, CBD almost completely normalized the AA-enhanced “pathological” lipogenesis of SZ95 sebocytes (Figure ​ (Figure1G). 1 G). This suggests that CBD may primarily normalize both quantitatively and qualitatively excessive and abnormal lipid production induced by acne-promoting stimuli.

CBD decreases proliferation, but not the viability, of human sebocytes both in vitro and ex vivo.

Besides the above lipostatic action, another desired effect of a proper anti-acne agent would be to inhibit the unwanted growth of sebocytes (2, 27, 28). Of great importance, proliferation of SZ95 sebocytes was significantly reduced in the presence of CBD (1–10 μM) (Figure ​ (Figure2A). 2 A). It should be noted, however, that CBD did not suppress the cell count below the “starting” number (measured at day 1), arguing for a “pure” antiproliferative effect. Indeed, the lack of its effects on the count of viable cells was further verified by showing that these concentrations of CBD did not decrease cellular viability or induce either apoptosis or necrosis of SZ95 sebocytes (Figure ​ (Figure2, 2 , B and C). Notably, administration of 50 μM CBD evoked apoptosis-driven cytotoxicity and, hence, led to decreased lipogenesis (Supplemental Figure 2, A–C). Likewise, elongated application of 10 μM CBD (6-day treatments) also decreased cell number and lipogenesis (Supplemental Figure 2, D and E).

(A) CyQUANT proliferation assay after 72-hour treatments. *P < 0.05, ***P < 0.001 compared with the 72-hour vehicle control. The solid line indicates the level of the 24-hour vehicle control. (B) MTT assay. Viability of sebocytes following 48-hour treatments. (C) Cell death [DilC1(5) and SYTOX Green double labeling] assays after 24-hour treatments. (AC) Results are expressed as the percentage of the vehicle control (mean ± SEM of 4 independent determinations). The solid line indicates 100%.Two additional experiments yielded similar results. (DG) hSOC of (D) control, (E) 10 μM CBD, (F) 30 μM AEA, and (G) 30 μM AEA plus CBD 10 μM (14 days; sebum: Oil Red O staining, red; nuclei: hematoxylin, blue). Scale bars: 50 μm. (H) Statistical analysis of the lipid production on 4 histological sections per group. Results are expressed as mean ± SEM. **P < 0.01. (I) Statistical analysis of the number of MKI67 + cells as compared with the number of DAPI + cells on 2 histological sections per group (hSOC; 48 hours). **P < 0.01 compared with the vehicle control. Results are expressed as mean ± SEM.

Clinically, the key question is whether the above in vitro observations could be translated into significant sebostatic (i.e., lipostatic and antiproliferative) effects of CBD on human sebaceous glands in situ. To explore this on the preclinical level, the full-thickness hSOC technique (20) was used. These hSOC assays, which mimic the human sebaceous gland function in vivo as closely as this is currently possible on the ex vivo level, clearly demonstrated that application of CBD completely prevented the lipogenic action of AEA in situ and, in line with our long-term in vitro observations (Supplemental Figure 2E), decreased basal lipogenesis as well (Figure ​ (Figure2, 2 , D–H). Likewise, CBD markedly suppressed the expression of the proliferation marker MKI67 (Figure ​ (Figure2I). 2 I). This suggests that CBD may also operate as a potent sebostatic agent in vivo when tested in appropriate clinical trials.

CBD exerts universal antiinflammatory actions.

We additionally found that CBD also prevented the “pro-acne” LA-T combination from elevating the expression of TNFA (Figure ​ (Figure3A), 3 A), a key cytokine in the pathogenesis of acne vulgaris (2, 24–30). These data suggested that CBD may exert antiinflammatory actions on human sebocytes (as had already been demonstrated for CBD in several other experimental models, such as diabetes, rheumatoid arthritis, etc.) (31). Therefore, in order to confirm the putative universal antiinflammatory action of the CBD on human sebocytes, we next assessed its effects by modeling both Gram-negative infections (applying the TLR4 activator LPS) and Gram-positive infections (using the TLR2 activator lipoteichoic acid [LTA]). We found that CBD completely prevented the above treatments from elevating TNFA expression (Figure ​ (Figure3). 3 ). Moreover, CBD also normalized LPS-induced IL1B and IL6 expression (Figure ​ (Figure3B) 3 B) (expression of these 2 cytokines was found not to be modulated by 24-hour LA-T or LTA treatment; data not shown). Taken together, these results strongly suggest that CBD’s universal sebostatic action is accompanied by substantial antiinflammatory effects, which would be very much desired in the clinical treatment of acne vulgaris (1, 2, 24–30).

(A) TNFA mRNA expression following 24-hour “pro-acne” lipogenic and TLR agonist treatments with or without CBD. *P < 0.05 compared with the corresponding CBD-free treatments. (B) IL1B, IL6, and TNFA mRNA expression following 24-hour LPS treatment with or without CBD. *P < 0.05, **P < 0.01, ***P < 0.001 compared with the corresponding CBD-free treatments. (A and B) Data are presented using the ΔΔCT method; GAPDH-normalized mRNA expression of the vehicle control was set as 1 (solid line). Data are expressed as mean ± SD of 3 independent determinations. Two additional experiments yielded similar results.

Sebostatic (i.e., lipostatic and antiproliferative), but not antiinflammatory, actions of CBD are mediated by the activation of transient receptor potential vanilloid-4 ion channels.

Next, we dissected the molecular mechanism(s) that underlie the remarkable lipostatic effects of CBD. As expected, neither CB1- nor CB2-specific antagonists (AM251 and AM630) were able to antagonize the lipid synthesis-inhibitory action of CBD (Supplemental Figure 3); hence, alternative options had to be considered.

First, we studied the effects of CBD on the ionic currents of SZ95 sebocytes. Using whole-cell patch-clamp configurations, membrane currents were elicited by voltage ramp protocols (Figure ​ (Figure4, 4 , A and B) and then normalized to cell membrane capacitance at two different potentials, i.e., at –90 and +90 mV (Figure ​ (Figure4C). 4 C). CBD (10 μM) induced a mostly outwardly rectifying current and a positive shift in the reversal potential, arguing for the activation of certain cation channels upon CBD application.

(A) Representative current-voltage traces of patch-clamp measurement of sebocytes using conventional whole-cell configuration with or without 10 μM CBD. (B) CBD-induced differential current (i.e., CBD minus control). (C) Averaged current densities measured at –90 mV and +90 mV of 7 cells. Results are expressed as mean ± SEM. **P < 0.01 compared with control.

It is well known that various cannabinoids can modulate the activity of certain transient receptor potential (TRP) channels, collectively referred to as “ionotropic cannabinoid receptors” (32–37). Moreover, we have shown recently that activation of TRP vanilloid-1 (TRPV1) on SZ95 sebocytes by capsaicin also exerts potent lipostatic actions (38). Therefore, we first systematically explored these candidate “CBD targets.”

We found that SZ95 sebocytes express TRPV1, TRPV2, and TRPV4 both at the mRNA and protein levels (Supplemental Figure 4, A–C). Among these TRP channels, TRPV4 showed the highest mRNA levels by far (expression of TRPA1 and TRPM8 was below the detection limit; data not shown).

Since the 3 identified TRPs are nonselective cation channels that are most permeable to Ca 2+ (39), we studied the effects of CBD on the calcium homeostasis of the sebocytes. Using a fluorescent Ca 2+ -imaging technique, we found that CBD significantly increased the intracellular calcium concentration ([Ca 2+ ]IC) of SZ95 sebocytes (Figure ​ (Figure5, 5 , A and B). This action was equally antagonized by (a) the decrease of the extracellular Ca 2+ concentration ([Ca 2+ ]EC); (b) the nonspecific TRP channel blocker ruthenium red; and, of great importance, (c) the TRPV4-specific antagonist HC067047 (HC) (Figure ​ (Figure5, 5 , A and B). We have also shown that the suppression of [Ca 2+ ]EC or the coapplication of HC also prevented the lipostatic action of CBD (Figure ​ (Figure5C); 5 C); notably, the TRPV4 antagonist alone did not affect basal lipid synthesis (Supplemental Figure 5).

To further confirm the functional expression of TRPV4 on human sebocytes, the TRPV4-specific ultrapotent agonist GSK1016790A (GSK) was applied. The agonist evoked membrane currents, which were prevented by the specific TRPV4 antagonist HC (Supplemental Figure 6, A and B), indicating that TRPV4 channels are indeed functionally expressed in human sebocytes. Moreover, GSK mimicked both the CBD-induced [Ca 2+ ]IC elevations (Supplemental Figure 6, C and D) and CBD’s lipostatic actions (Figure ​ (Figure5C). 5 C). Since the CBD-evoked lipostatic effects and the induced Ca 2+ signals were not influenced by the TRPV1-specific antagonists, capsazepine (Supplemental Figure 7, A–C) or AMG 9810 (data not shown), these electrophysiological, Ca 2+ -imaging and cellular physiology data collectively argued for the selective involvement of TRPV4 (but not of TRPV1) in mediating the effects of CBD.

To further validate this concept, knockdown of TRPV1, TRPV2, and TRPV4 by RNA interference (RNAi) was used (quantitative “real-time” PCR [Q-PCR] and Western blot analyses verified the successful silencing of the targeted TRPVs; Supplemental Figure 8, A–F). We showed that neither TRPV1 nor TRPV2 silencing significantly influenced the lipostatic action of CBD (Supplemental Figure 9, A and B). In contrast, TRPV4-specific “knockdown” was able to prevent this effect of CBD (Figure ​ (Figure5D) 5 D) as well as the increase of [Ca 2+ ]IC (Supplemental Figure 10) and the lipid-lowering action of the TRPV4-specific activator GSK (Figure ​ (Figure5E). 5 E). Collectively, these data unambiguously confirm that CBD activates TRPV4 and that this ion channel selectively mediates its lipostatic action.

Interestingly, we also showed that, similar to the lipostatic action, antagonism of TRPV4 was able to significantly prevent the antiproliferative effect of CBD (Figure ​ (Figure6A). 6 A). However, quite surprisingly, antiinflammatory actions of CBD were not affected by the antagonist (Figure ​ (Figure6B); 6 B); these latter findings suggested that these antiinflammatory actions might be a TRPV4-independent process.

(A) CyQUANT proliferation assay after 72-hour treatments. *P < 0.05 compared with the vehicle control. # P < 0.05. The solid line indicates the level of the 24-hour vehicle control. Dashed line indicates the level of the 72-hour vehicle control. Results are expressed as the percentage of the 24-hour vehicle control (mean ± SEM of 4 independent determinations). (B) TNFA mRNA expression following 24-hour LPS treatments with or without CBD and HC. *P < 0.05 compared with the vehicle control; # P < 0.05 compared with the CBD-free LPS-treated group. Data are presented using the ΔΔCT method; peptidyl-prolyl isomerase A–normalized (PPIA-normalized) TNFA mRNA expression of the vehicle control was set as 1. Data are expressed as mean ± SD of 3 independent determinations. Two additional experiments yielded similar results. (C) Validation of the key microarray results. mRNA expression of various target genes following 24-hour CBD treatments with or without HC. **P < 0.01, ***P < 0.001 compared with the vehicle control. ### P < 0.001. Data are presented using the ΔΔCT method; PPIA-normalized mRNA expression of the vehicle control was set as 1 (solid line). Data are expressed as mean ± SD of 3 to 6 independent determinations. Two additional experiments yielded similar results.

Sebostatic action of CBD is mediated by TRPV4-dependent interference with the ERK1/2 MAPK pathway and downregulation of nuclear receptor interacting protein-1.

To dissect the intracellular signaling pathways that underlie the above effects, we first investigated the putative participation of several kinases (i.e., PKC isoforms, PI3K, PKA) as well as calcineurin in mediating the lipostatic effects of CBD. Notably, inhibition of activities of these molecules had no effect on the lipostatic activity of CBD (Supplemental Figure 11, A and B).

Then, in order to identify target genes and pathways regulated (directly or indirectly) by CBD, genome-wide microarray analyses were performed on 3 independent sets of control and CBD-treated SZ95 sebocytes (10 μM CBD for 24 hours). Gene set enrichment analysis (GSEA) (40–42) of the microarray results revealed that numerous mitosis and cell cycle (e.g., “mitosis,” “G2/M transition,” “cell cycle,” etc.), inflammation (e.g., “cytokine production,” “cytokine biosynthetic process,” “TLR9 pathway,” “positive regulation of IκB kinase NF-κB cascade,” etc.), and lipid synthesis–related (“acyltransferase activity,” “lipid biosynthetic process,” “positive regulation of MAPK activity,” etc.) gene sets were identified among the downregulated ones, confirming our previous findings on the complex anti-acne effects of CBD. Moreover, downregulation of some “acne-related” gene sets (e.g., “IGF-1 pathway” and “mTOR pathway”) (2, 43) also argued for the putative in vivo anti-acne efficiency of CBD. Further, upregulation of the “calcium signaling pathway” gene set confirmed the involvement of (TRPV4-dependent) calcium signaling (detailed results of GSEA are available in Supplemental Excel files 1 and 2).

During further data processing, Biological Networks Gene Ontology (BiNGO) analysis (44, 45) was also performed (see Supplemental Excel files 3 and 4; the hierarchy of the different gene ontology terms enriched among the downregulated and upregulated genes is summarized in Supplemental Figures 12 and 13, respectively). In line with our previous results, this method also highlighted that CBD exerted “anti-differentiating” effects on sebocytes (terms like “negative regulation of fat cell differentiation” and “negative regulation of fatty acid biosynthetic process” were found to be enriched among the upregulated genes).

Although these analyses further confirmed our previous findings on the complex anti-acne effects of CBD, we still aimed to recognize target genes that might be involved in mediating the different anti-acne modalities and/or might further strengthen the putative in vivo efficiency of CBD. Therefore, using rigid exclusion criteria (at least 2-fold changes in the corresponding expression levels equidirectional changes in all cases, and global, corrected P < 0.05), we found that 80 genes were significantly downregulated, whereas 72 genes were significantly upregulated by CBD treatment (microarray results are accessible through GEO series accession number > GSE57571; downregulated and upregulated genes, together with their averaged fold changes, are summarized in Supplemental Tables 1 and 2). By using Q-PCR, we have confirmed that, following CBD treatment, expression of Rho GTPase-activating protein 9 (ARHGAP9, an endogenous inhibitor of the prolipogenic ERK signaling) (46) was upregulated, whereas the proliferation marker MKI67 was downregulated (Figure ​ (Figure6C). 6 C). (This latter result perfectly confirmed our findings obtained in hSOC experiments [Figure ​ [Figure2I].) 2 I].) Moreover, also in line with our previous findings, we found that TRPV4 antagonism could successfully prevent both alterations (Figure ​ (Figure6 6 C).

It is well known that activation of the ERK1/2 MAPK pathway plays a crucial role in the regulation of cellular proliferation (47). Furthermore, we have demonstrated recently that this pathway is involved in mediating the “prolipogenic” action of AEA on human sebocytes (12). Considering that administration of CBD led to opposing cellular effects (i.e., decreased lipogenesis and proliferation) and upregulation the ERK inhibitor ARHGAP9, we hypothesized that CBD might inhibit MAPK activation. Indeed, AEA treatment was able to activate the ERK1/2 MAPK cascade (as monitored by assessing the level of phosphorylated ERK1/2 [P-ERK1/2]), an effect that was completely abrogated by the coadministration of CBD (Figure ​ (Figure7A). 7 A). In a perfect agreement with our previous data (Figure ​ (Figure5, 5 , C–E, and Figure ​ Figure6, 6 , A and C), this interference was found to be TRPV4 dependent, since the specific antagonist HC was able to fully prevent the effect of CBD (Figure ​ (Figure7A). 7 A). This, again, confirmed the crucial role of TRPV4 activation in initiating the lipostatic and antiproliferative signaling cascade(s) of CBD.

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Lipostatic effects of CBD are mediated by TRPV4-dependent inhibition of the prolipogenic ERK1/2 signaling and downregulation of NRIP1.

(A) Western blot analysis of lysates of SZ95 sebocytes treated with 30 μM AEA, 10 μM CBD, and 1 μM HC for 5 minutes. Numbers on the OD row indicate the optical density of the P-ERK1/2 bands normalized to the corresponding ERK1/2 signals. (B) Quantitative determination of neutral lipid synthesis (Nile Red staining; 24-hour treatments started at day 3 after transfection). **P < 0.01, ***P < 0.001 compared with the scrambled (SCR) control group. “siNRIP1a” and “siNRIP1b” mark 2 different siRNA constructs against NRIP1. Data are expressed as the percentage of the SCR vehicle control (mean ± SEM of 4 independent determinations). The solid line indicates 100%.One additional experiment yielded similar results.

We have also demonstrated that expression of nuclear receptor interacting protein-1 (NRIP1, also known as RIP140; a corepressor essential for triglyceride storage in adipose tissue) (48) was downregulated in a TRPV4-dependent manner (Figure ​ (Figure6C). 6 C). We have shown that silencing of NRIP1 (validated by Q-PCR and Western blotting; Supplemental Figure 14, A and B) mimicked the lipostatic effect of CBD (Figure ​ (Figure7B), 7 B), suggesting that downregulation of NRIP1 is indeed an important final effector of the lipid synthesis-inhibitory activity of CBD.

Antiinflammatory action of CBD is mediated by upregulation of tribbles homolog 3 and inhibition of the NF-κB pathway.

Our microarray data have also highlighted the putative involvement of several innate immunity/inflammation-related genes in mediating the antiinflammatory action of CBD (Supplemental Tables 1 and 2). By using Q-PCR, we confirmed that expression of LL-37 cathelicidin (a key antimicrobial peptide expressed by and shown to be active in human sebocytes) (49) and tribbles homolog 3 (TRIB3, also known as SINK; a negative regulator of proinflammatory NF-κB signaling) (50) was upregulated by CBD. Importantly (again, in line with our previous results [Figure ​ [Figure6B]), 6 B]), these CBD-induced gene expression changes were not prevented by the coadministration of the TRPV4 antagonist HC (Figure ​ (Figure6C). 6 C). When assessing the functional role of TRIB3, we found that, after its selective silencing (Supplemental Figure 15, A and B), CBD was unable to exert its antiinflammatory action to prevent LPS-induced IL1B and IL6 upregulation (Figure ​ (Figure8A); 8 A); in contrast, its lipostatic activity was not altered (Supplemental Figure 15C).

Antiinflammatory actions of CBD are coupled to A2a receptor-dependent upregulation of TRIB3 and subsequent inhibition of the P65-NF-κB signaling.

TRIB3 is known to inhibit the NF-κB pathway (50), and, furthermore, CBD has already been reported to exert its antiinflammatory actions via inhibition of the NF-κB signaling (51). Importantly, we found that CBD cotreatment indeed prevented the LPS-induced phosphorylation (hence inactivation) of the inhibitory IκBα and phosphorylation (hence activation) of the p65 (RelA) NF-κB isoform (Figure ​ (Figure8B). 8 B). These data indicate that, irrespective of the investigated cell type, interference with the NF-κB pathway could be an important mechanism in the development of the antiinflammatory actions of CBD. It should also be noted that TRPV4 antagonism exerted only negligible effects on the action of CBD (Figure ​ (Figure8B), 8 B), again confirming that antiinflammatory activity of CBD is a TRPV4-independent process.

CBD induces a novel (A2a adenosine receptor→cAMP→TRIB3⊣NF-κB) antiinflammatory pathway.

Finally, we aimed at identifying the target molecule of CBD, which, via the upregulation of TRIB3, mediates the antiinflammatory action of the phytocannabinoid. Since previous data suggested that elevation of the intracellular cAMP level is one of the possible inducers of TRIB3 activation/upregulation (52), we have investigated the effects of CBD on the cAMP level. CBD treatment indeed elevated the intracellular cAMP concentration of the sebocytes (Figure ​ (Figure8C), 8 C), arguing that a Gs protein–coupled receptor might be the primary target of CBD. A previous finding that, in a murine model of acute lung injury, the Gs protein–coupled A2a adenosine receptor was found to mediate the antiinflammatory actions of CBD (53) made this receptor a very probable target in our system as well. Indeed, we found that the A2a receptor was expressed by human sebocytes both at the mRNA and protein levels (Supplemental Figure 16, A–C). In addition, we have also shown that application of a specific A2a receptor antagonist, ZM241385 (ZM), was able to significantly prevent the upregulation of TRIB3 by CBD (Figure ​ (Figure8D). 8 D). Moreover, ZM also suppressed the antiinflammatory effect of the phytocannabinoid as well as the CBD-evoked inhibition of LPS-induced NF-κB activation (Figure ​ (Figure8, 8 , E and F). These intriguing findings collectively argued that activation of the “A2a receptor→cAMP→TRIB3⊣NF-κB” axis indeed plays a crucial role in mediating the antiinflammatory actions of CBD.

Discussion

In this study, we provide the first evidence that the nonpsychotropic phytocannabinoid CBD, which is already applied in clinical practice (16), exerted a unique “trinity of cellular anti-acne actions.” Namely, CBD, without compromising viability (Figure ​ (Figure2, 2 , B and C), (a) normalized the pathologically elevated lipogenesis induced by “pro-acne” agents, both in a quantitative and qualitative manner (universal lipostatic effect; Figure ​ Figure1); 1 ); (b) suppressed cell proliferation (antiproliferative effect; Figure ​ Figure2A); 2 A); and (c) prevented the actions of TLR activation or “pro-acne” agents to elevate proinflammatory cytokine levels (universal antiinflammatory effect; Figure ​ Figure3). 3 ). Furthermore, we have shown that sebostatic actions of CBD also developed under “in vivo–like” conditions (hSOC; Figure ​ Figure2, 2 , D–I).

Besides the discussed “sebocyte-specific” steps of the pathogenesis of acne, promisingly targeted by the “cellular anti-acne trinity” of CBD, one should also keep in mind that there are additional factors, which contribute to the progression of the disease: the infundibular hyperproliferation/hyperkeratinization, leading to comedogenesis and subsequent overgrowth of “acnegenic” Propionibacterium acnes strains (2). It is very important to note that, based on the literature, administration of CBD holds out the promise to target these factors as well. Indeed, CBD was shown to inhibit proliferation of hyperproliferative keratinocytes (54), and it was demonstrated to possess remarkable antibacterial activity (55). Although its efficiency against “acnegenic” Propionibacterium acnes strains is not yet investigated, one can speculate that its putative indirect antibacterial activity (mediated by the upregulation of the expression of the antimicrobial peptide LL-37 cathelicidin [Supplemental Table 2 and Figure ​ Figure6C]) 6 C]) could be further supported by direct bactericide effects, arguing that CBD might be very likely to behave as a potent anti-acne agent in vivo.

Given that sebum production is the result of holocrine secretion, the amount of sebum produced is at least as dependent on the proliferative activity of basal layer sebocytes in the sebaceous gland as on the amount of lipogenesis that individual sebocytes engage in (27, 28). Therefore, the novel and significant antiproliferative activity of CBD on human sebocytes in vitro and ex vivo documented here (Figure ​ (Figure2, 2 , A and I) is expected to greatly reduce sebum production in vivo. Moreover, it is also important to emphasize that, clinically, it is highly desirable that basal sebogenesis and viability of sebocytes are unaffected (Figure ​ (Figure1, 1 , A–C, and Figure ​ Figure2, 2 , A–C) by CBD (at least in the noncytotoxic concentrations and after short-term treatments; Supplemental Figure 2, A–E), since a sufficient level of sebum production is a critical factor for maintaining proper function of the epidermal barrier, one of the central components of skin homeostasis (56).

CBD has already been shown to activate (e.g., certain TRP channels, α1 and 5-HT1a receptor, etc.), antagonize (e.g., TRPM8 and 5-HT3 receptor as well as “classical” [CB1 and CB2] and “novel” [GPR55] cannabinoid receptors, etc.), or allosterically modulate (e.g., μ- and δ-opioid receptors, etc.) the activity of a plethora of different receptors and, furthermore, to influence various other cellular targets (e.g., cyclooxygenase and lipoxygenase enzymes, fatty acid amide hydrolase, eCB membrane transporter, phospholipase A2, voltage-dependent anion channel 1, etc.) (15, 32–37, 57–60). Therefore, exploration of its exact mechanism of action appeared to be a great challenge. The fact that we have shown previously that activation of TRPV1 can evoke similar lipostatic effects (38) as those found for CBD (Figure ​ (Figure1 1 and Figure ​ Figure2, 2 , D–H), together with our present findings that CBD induced membrane currents on sebocytes (Figure ​ (Figure4), 4 ), prompted us to first investigate the role of TRP channels in mediating the above anti-acne modalities. We discovered that the lipostatic and antiproliferative effects of CBD were mediated by the activation of TRPV4 (and not TRPV1 or TRPV2) ion channels (Figures ​ (Figures5, 5 , C–E, and Figure ​ Figure6A) 6 A) and the concomitant increase in [Ca 2+ ]IC. Actually, the “negative regulation” of lipogenesis by the elevation of [Ca 2+ ]IC is not unprecedented, since it has already been described in sebocytes (38) as well as in adipocytes (61, 62). It is also important to note that, within the confines of another study, we have shown that extracellular Ca 2+ plays an important negative regulatory role in the sebaceous lipogenesis (C.C. Zouboulis et al., unpublished observations). Of further importance, we have also shown that the antiinflammatory activity of CBD is a TRPV4-independent process (Figure ​ (Figure6 6 B).

Importantly, our data are in perfect agreement with the recent findings of De Petrocellis et al. (37). Using heterologous expression systems, they demonstrated that CBD is a potent but less efficacious activator of rat TRPV4 (as compared with the “classical” agonists or certain other phytocannabinoids, such as cannabichromene [CBC] or cannabidivarin [CBDV]). Although the possibility that CBD might be a more efficacious activator of human TRPV4 than of rat TRPV4 should also be taken into consideration; preliminary data of our recently started assessment of the putative anti-acne effects of other phytocannabinoids also suggest that CBC and CBDV possess an even more pronounced lipostatic efficiency than CBD, which further argues for the central role of TRPV4 (A. Oláh et al., unpublished observations).

In order to identify additional downstream targets, genome-wide microarray experiments were performed on 3 independent sets of control and CBD-treated (10 μM for 24 hours) sebocytes. GSEA (40–42) and BiNGO analysis (44, 45) of the microarray results uniformly confirmed our results, arguing for complex anti-acne actions upon CBD administration, as indicated by downregulation of inflammation (e.g., “cytokine production”), lipid synthesis (e.g., “lipid biosynthetic process” and “positive regulation of MAPK activity”), proliferation-related (e.g., “mitosis” and “G2/M transition”), and “general pro-acne” (e.g., “mTOR pathway” and “IGF-1 pathway”) (2, 43) gene sets and BiNGO terms (Supplemental Excel files 1–4 and Supplemental Figures 12 and 13).

Besides the above results, microarray analyses also revealed that levels of 80 genes were downregulated upon CBD treatment, whereas expression of 72 genes was upregulated upon CBD treatment, among which multiple potential “anti-acne” effectors were identified (Supplemental Tables 1 and 2). Q-PCR validation of the most promising target genes revealed that (in agreement with our cell physiology data) expression of lipid synthesis–related (NRIP1 and ARHGAP9) and proliferation-related (MKI67) genes was altered in a TRPV4-dependent manner, whereas changes in the expression of “inflammation” genes were found to be TRPV4 independent (Figure ​ (Figure6C). 6 C). Moreover, alterations of ARHGAP9 expression (a known endogenous inhibitor of ERK signaling) (46) suggested that inhibition of the prolipogenic MAPK pathway (12) might play a role in mediating the lipostatic effects of CBD. Indeed, we found that CBD inhibited AEA-induced (prolipogenic) (12) ERK1/2 phosphorylation in a TRPV4-dependent manner (Figure ​ (Figure7A), 7 A), confirming again the crucial role of TRPV4 in mediating the action of CBD.

We also silenced another “lipid-regulating gene” (i.e. NRIP1) (Supplemental Figure 14, A and B). As expected (48), knockdown of NRIP1 was able to mimic the lipostatic effect of CBD (Figure ​ (Figure7 7 B).

Next, we aimed at revealing the signaling pathway of the antiinflammatory actions. Thorough assessment of the microarray data highlighted the putative role of TRIB3, a known inhibitor of proinflammatory NF-κB signaling (50). In addition, inhibition of NF-κB signaling plays a crucial role in the development of CBD-mediated antiinflammatory actions in other systems (51). RNAi-mediated selective gene silencing of TRIB3 in human sebocytes (Supplemental Figure 15, A and B) fully abrogated the ability of CBD to inhibit LPS-induced proinflammatory responses (Figure ​ (Figure8A). 8 A). Although a previous study would have suggested it (63), interestingly, TRIB3 was found not to participate in mediating the lipostatic effects of CBD in sebocytes (Supplemental Figure 15C).

It is also noteworthy that TRIB3 has been identified recently as a potent phytocannabinoid target gene (64–66). These results, together with our data presented here, strongly argue for the key participation of TRIB3 in mediating cellular effects of cannabinoids.

Although CBD-dependent upregulation of its several known target genes, such as activating transcription factor 4, asparagine synthetase, cation transport regulator-like 1, and DNA-damage-inducible transcript 3 (refs. 66, 67, and Supplemental Tables 1 and 2), also argued for the activation of a TRIB3-dependent signaling pathway, to further strengthen the “TRIB3-hypothesis,” we have also investigated the effects of CBD on one of the major cellular targets of TRIB3, i.e., NF-κB. As expected (51), CBD was able to inhibit LPS-induced NF-κB activation (again, in a TRPV4-independent manner; Figure ​ Figure8B), 8 B), which can fully explain its previously demonstrated antiinflammatory actions.

Finally, we aimed at identifying the upstream signaling of the TRIB3 activation/upregulation by CBD. We found that CBD elevated the level of cAMP (a known upstream regulator of TRIB3) (ref. 52 and Figure ​ Figure8C), 8 C), highlighting the putative role of a Gs-coupled receptor in initiating its antiinflammatory actions. We also demonstrated that sebocytes express Gs-coupled A2a receptors (which have already been shown to mediate antiinflammatory actions of CBD) (ref. 53 and Supplemental Figure 16, A–C). Further, the specific A2a antagonist (ZM) was able to prevent upregulation of TRIB3 upon CBD treatment (Figure ​ (Figure8D). 8 D). Then, we attempted to confirm the functional presence of the putative antiinflammatory A2a receptor→cAMP→TRIB3′ΔΤNF-κB axis. We found that coadministration of ZM abrogated the antiinflammatory action of CBD (Figure ​ (Figure8E). 8 E). Moreover, we were also able to show that it abolished the NF-κB–inhibitory action of CBD (Figure ​ (Figure8F). 8 F). Taken together, these data strongly argue that A2a receptor might be the primary orchestrator of the antiinflammatory actions of CBD. It should also be noted that, according to the data published by Carrier et al. (68), CBD-mediated activation of A2a receptor is very likely to be an indirect action, realized by the primary inhibition of the equilibrative nucleoside transporter(s) (e.g., ENT1, which mediates adenosine uptake of the cells) and the subsequently elevated “adenosine tone.”

Collectively, our data introduce the phytocannabinoid CBD as a potent “universal” anti-acne agent, possessing a unique “triple anti-acne” profile (Figure ​ (Figure9). 9 ). Multiple human studies have already investigated the safety of CBD (13, 14). Furthermore, it is already in use in many countries in clinical practice without any significant side effects (Sativex) (16). This is especially promising, because the currently available, most effective anti-acne agent, isotretinoin, is known to cause serious side effects (2, 69, 70). These data, together with our current findings, point to a promising, cost-effective, and, likely, well-tolerated new strategy for treating acne vulgaris, the most common human skin disease.