Sulforaphane activates the transcription factor Nrf2 and induces phase-2 and antioxidant enzymes in human keratinocytes

Experimental Dermatology 2009 2009 Jun 23.
Sulforaphane but not ascorbigen, indole-3-carbinole and ascorbic acid activates the transcription factor Nrf2 and induces phase-2 and antioxidant enzymes in human keratinocytes in culture.
Wagner AE, Ernst I, Iori R, Desel C, Rimbach G.

Institute of Human Nutrition and Food Science, Christian-Albrechts-University, Kiel, Germany.

Nrf2 is a basic leucine zipper transcriptional activator essential for the coordinated transcriptional induction of phase-2 and antioxidant enzymes. Brassica vegetables contain phytochemicals including glucoraphanin, the precursor of sulforaphane (SFN) and glucobrassicin, the precursor of indole-3-carbinole (I3C) and ascorbigen (ABG). The degradation products SFN, I3C and ABG may be capable of inducing cytoprotective genes in skin. In this study, we tested the potency of SFN, ABG and I3C in affecting Nrf2-dependent gene expression in human keratinocytes in culture. SFN but not ABG and its precursors I3C and ascorbic acid induced Nrf2 dependent gene expression at a relatively low concentration (5 mumol/l). Induction of Nrf2 due to SFN was accompanied by an increase in mRNA and protein levels of NADPH quinone oxidoreductase 1, heme oxygenase 1 and gamma-glutamylcysteine-synthetase. Furthermore, SFN elevated cellular glutathione levels and antagonized tumor necrosis factor-alpha-induced NFkappaB transactivation. Therefore, SFN treatment may present a strategy for enhancing the cellular defense mechanisms in skin.

This is probably the most important study I've seen in 6-months. It may confirm a theory I've had recently about Sulforaphane, which is that it possibly could transcriptionally alter the antioxidant protection in the hair follicle. In male pattern baldness, there is a lower level of glutathione and other protectants. Nrf2 is a very relevant transcription factor which regulates oxidant stress through the primary defense systems that can protect the hair follicle.

And at least in the case of prostate tissue it can repress the androgen receptor (Mol Cancer Ther. 2009 Jul;8(7):1946-54.).

Based on this and a few other studies involving Sulforaphane and keratinocytes, this maybe a key nutrient in those with recalcitrant scalp inflammation.

Having tried this before, it seemed to be helping, but I did not use it long enough to be absolutely sure. This is going back into my shopping chart

http://www.iherb.com/Jarrow-Formulas-Activated-Selenium-200-mcg-60-Capsules/169?at=0

CS--

I've been using Brocomax thrice daily for a couple of months and feel like it has been a great addition to my health protocol. Would you recommend selenamax over brocomax, or is it just your recommendation based on dollar value? Thanks.

Gibson - Yes, both the economic value and the selenium which complements phase II detoxification and Sulforaphane.

Of course the Broccomax is much higher in Sulforaphane and since Helicobacter pylori plays at least a contributory role in some skin diseases the extra amount won't hurt at all. But I just love the price of this selenuim combination. It's like buying a great form of Selenium with a little free Broccomax thrown in.

Gibson - I forgot to mention that research wise, there's too little known about dosing, so whatever experience you have be would be of great interest. So far I've just used the Selenium version with the 9 mg of Broccomax and after that took a month off. And for whatever reason I have a good feeling about it and this new research makes me want to try it for much longer.

Are you just taking it once a day CS? Is it okay to be taking that much Selenium?

hapyman - Yes, just once per day.

CS--

When I added it--this was around the same time I added BCM--I felt a huge boost. I actually got a little excited at one point and lined my favorite supplements up for a photo shoot (inflammation totally, completely eradicated, profound sense of well-being).

As these things go, the difference becomes the norm, so I can't tell otherwise; other than to say, I have no inflammation and feel very healthy.

My most recent point of attack has been cortisol, at your suggestion, and I am doing sensoril 3x a day, and just started PS, literally yesterday.

Generally, I dose 3x a day, just a practice. Depending on the supplement, I may exclude it from evening or morning. Brocomax has been perfectly tolerated by me at that dose, but may be unessecarily excessive.

CausticSymmetry wrote:I forgot to mention that research wise, there's too little known about dosing, so whatever experience you have be would be of great interest.

Hey CausticSymmetry,

FWIW, while not related to hair loss, I've taken the selenium version that you linked to at 1 pill per day and I'm just barely able to get by without my allergy symptoms being too bad.

However, the Broccomax at 1 pill per day (without selenium) is awesome.

I believe I'll be taking Sulforaphane for the rest of my life based on how well it works for me.

Ever since the time of this thread, I've been taking about 9 milligrams of Sulforaphane per day. I don't think I've had a bad hair day since. I'm wondering if anyone else has had this experience. Below are a few related studies.

J Nutr Biochem. 2009 Jul 1.
Sulforaphane suppresses ultraviolet B-induced inflammation in HaCaT keratinocytes and HR-1 hairless mice.
Shibata A, Nakagawa K, Yamanoi H, Tsuduki T, Sookwong P, Higuchi O, Kimura F, Miyazawa T.

Food and Biodynamic Chemistry Laboratory, Graduate School of Agricultural Science, Tohoku University, Sendai 981-8555, Japan.

Ultraviolet B (UVB) irradiation induces skin damage and inflammation. One way to reduce the inflammation is via the use of molecules termed photochemopreventive agents. Sulforaphane (4-methylsulfinylbutyl isothiocyanate, SF), which is found in cruciferous vegetables, is known for its potent physiological properties. This study was designed to evaluate the effect of SF on skin inflammation in vitro and in vivo. In in vitro study using immortalized human keratinocytes (HaCaT), UVB caused marked inflammatory responses [i.e., decrease of HaCaT viability and increase of production of an inflammatory marker interleukin-6 (IL-6)]. SF recovered the cell proliferation and suppressed the IL-6 production. These anti-inflammatory effects of SF were explained by its ability to reduce UVB-induced inflammatory gene expressions [IL-6, IL-1beta and cyclooxgenase-2 (COX-2)]. Because SF seems to have an impact on COX-2 expression, we focused on COX-2 and found that SF reduced UVB-induced COX-2 protein expression. In support of this, PGE(2) released from HaCaT was suppressed by SF. Western blot analysis revealed that SF inhibited p38, ERK and SAPK/JNK activation, indicating that the inhibition of mitogen-activated protein kinases (MAPK) by SF would attenuate the expression of inflammatory mediators (e.g., COX-2), thereby reducing inflammatory responses. Moreover, we conducted skin thickening assay using HR-1 hairless mice and found that UVB-induced skin thickness, COX-2 protein expression and hyperplasia were all suppressed by feeding SF to the mice. These results suggest that SF has a potential use as a compound for protection against UVB-induced skin inflammation.

Exp Dermatol. 2009 Jun 23.
Sulforaphane but not ascorbigen, indole-3-carbinole and ascorbic acid activates the transcription factor Nrf2 and induces phase-2 and antioxidant enzymes in human keratinocytes in culture.
Wagner AE, Ernst I, Iori R, Desel C, Rimbach G.

Institute of Human Nutrition and Food Science, Christian-Albrechts-University, Kiel, Germany.

Please cite this paper as: Sulforaphane but not ascorbigen, indole-3-carbinole, and ascorbic acid activates the transcription factor Nrf2 and induces phase-2 and antioxidant enzymes in human keratinocytes in culture. Experimental Dermatology 2009.Abstract: Nrf2 is a basic leucine zipper transcriptional activator essential for the coordinated transcriptional induction of phase-2 and antioxidant enzymes. Brassica vegetables contain phytochemicals including glucoraphanin, the precursor of sulforaphane (SFN) and glucobrassicin, the precursor of indole-3-carbinole (I3C) and ascorbigen (ABG). The degradation products SFN, I3C and ABG may be capable of inducing cytoprotective genes in skin. In this study, we tested the potency of SFN, ABG and I3C in affecting Nrf2-dependent gene expression in human keratinocytes in culture. SFN but not ABG and its precursors I3C and ascorbic acid induced Nrf2 dependent gene expression at a relatively low concentration (5 mumol/l). Induction of Nrf2 due to SFN was accompanied by an increase in mRNA and protein levels of NADPH quinone oxidoreductase 1, heme oxygenase 1 and gamma-glutamylcysteine-synthetase. Furthermore, SFN elevated cellular glutathione levels and antagonized tumor necrosis factor-alpha-induced NFkappaB transactivation. Therefore, SFN treatment may present a strategy for enhancing the cellular defense mechanisms in skin.

Planta Med. 2008 Oct;74(13):1526-39.
Nrf2 as a master redox switch in turning on the cellular signaling involved in the induction of cytoprotective genes by some chemopreventive phytochemicals.
Surh YJ, Kundu JK, Na HK.

National Research Laboratory of Molecular Carcinogenesis and Chemoprevention, College of Pharmacy, Seoul National University, Seoul, South Korea. surh@plaza.snu.ac.kr

A wide array of dietary phytochemicals have been reported to induce the expression of enzymes involved in both cellular antioxidant defenses and elimination/inactivation of electrophilic carcinogens. Induction of such cytoprotective enzymes by edible phytochemicals largely accounts for their cancer chemopreventive and chemoprotective activities. Nuclear factor-erythroid-2-related factor 2 (Nrf2) plays a crucial role in the coordinated induction of those genes encoding many stress-responsive and cytoptotective enzymes and related proteins. These include NAD(P)H:quinone oxidoreductase-1, heme oxygenase-1, glutamate cysteine ligase, glutathione S-transferase, glutathione peroxidase, thioredoxin, etc. In resting cells, Nrf2 is sequestered in the cytoplasm as an inactive complex with the repressor Kelch-like ECH-associated protein 1 (Keap1). The release of Nrf2 from its repressor is most likely to be achieved by alterations in the structure of Keap1. Keap1 contains several reactive cysteine residues that function as sensors of cellular redox changes. Oxidation or covalent modification of some of these critical cysteine thiols would stabilize Nrf2, thereby facilitating nuclear accumulation of Nrf2. After translocation into nucleus, Nrf2 forms a heterodimer with other transcription factors, such as small Maf, which in turn binds to the 5'-upstream CIS-acting regulatory sequence, termed antioxidant response elements (ARE) or electrophile response elements (EpRE), located in the promoter region of genes encoding various antioxidant and phase 2 detoxifying enzymes. Certain dietary chemopreventive agents target Keap1 by oxidizing or chemically modifying one or more of its specific cysteine thiols, thereby stabilizing Nrf2. In addition, phosphorylation of specific serine or threonine residues present in Nrf2 by upstream kinases may also facilitate the nuclear localization of Nrf2. Multiple mechanisms of Nrf2 activation by signals mediated by one or more of the upstream kinases, such as mitogen-activated protein kinases, phosphatidylionositol-3-kinase/Akt, protein kinase C, and casein kinase-2 have recently been proposed. This review highlights the cytoprotective gene expression induced by some representative dietary chemopreventive phytochemicals with the Nrf2-Keap1 system as a prime molecular target.

Pigment Cell Melanoma Res. 2008 Feb;21(1):79-88.
The significance of Nrf2 pathway in (photo)-oxidative stress response in melanocytes and keratinocytes of the human epidermis.
Marrot L, Jones C, Perez P, Meunier JR.

Department of Safety Research, Phototoxicity Unit, L'OREAL Research, Aulnay-Sous-Bois, France. lmarrot@rd.loreal.com

The expression of genes encoding antioxidant and/or phase 2 detoxifying enzymes can be enhanced in response to various environmental stresses. The main transcription factor involved in this response is nuclear factor erythroid 2-related factor 2 (Nrf2). Nrf2 activity is negatively regulated by the protein Kelch-like-Ech-associated-protein 1 (Keap1). While the roles of Nrf2 and phase 2 genes in chemoprevention of carcinogenesis have been well described; only few studies have dealt with their role in skin cancer. Normal human keratinocytes (NHK) and melanocytes (NHM) were treated by chemical inducers of the Nrf2 pathway or by small interfering RNAs (siRNA) used to knock down Keap1 mRNA. The above treatments resulted in significant stimulation of NQO-1 (NADPH-Quinone-Oxidoreductase 1) gene expression. GCL (gamma-Glutamyl-cysteinyl-ligase) gene was also induced but interestingly increased mRNA encoding the catalytic, heavy subunit GCLC was mainly stimulated in NHK, whereas the mRNA encoding the modifier, light subunit GCLM was mostly induced in NHM. HO-1 (Heme Oxygenase 1) gene induction was relatively strong in NHM, but generally absent in NHK, except when the cells were subjected to cytotoxic doses of the above chemicals. Exposure to solar UV (UVB + UVA, 300-400 nm) or to UVA alone (320-400 nm) confirmed this trend, but interestingly, at doses where cell growth reduction was comparable, UVA was generally more efficient than solar UV in inducing phase 2 genes. When siRNAs directed against Nrf2 were used, a strong down-regulation of NQO-1 expression was observed in both, NHM and NHK, whereas reduction of HO-1 expression was mainly detected in NHM. To our knowledge, this is the first study comparing phase 2 gene modulation in NHK and NHM. The results hereby presented should contribute to a better understanding of the molecular mechanisms involved in skin adaptation to environmental stress.

Arterioscler Thromb Vasc Biol. 2009 Sep 3.
Activation of Nrf2 in Endothelial Cells Protects Arteries From Exhibiting a Proinflammatory State.
Zakkar M, Van der Heiden K, Luong LA, Chaudhury H, Cuhlmann S, Hamdulay SS, Krams R, Edirisinghe I, Rahman I, Carlsen H, Haskard DO, Mason JC, Evans PC.

British Heart Foundation Cardiovascular Sciences Unit, National Heart and Lung Institute, Imperial College London, UK; the Department of Bioengineering, Imperial College London, UK; University of Rochester Medical Center, Rochester, NY; and the Department of Nutrition, University of Oslo, Norway.

OBJECTIVE: Proinflammatory mediators influence atherosclerosis by inducing adhesion molecules (eg, VCAM-1) on endothelial cells (ECs) via signaling intermediaries including p38 MAP kinase. Regions of arteries exposed to high shear stress are protected from inflammation and atherosclerosis, whereas low-shear regions are susceptible. Here we investigated whether the transcription factor Nrf2 regulates EC activation in arteries. METHODS AND RESULTS: En face staining revealed that Nrf2 was activated in ECs at an atheroprotected region of the murine aorta where it negatively regulated p38-VCAM-1 signaling, but was expressed in an inactive form in ECs at an atherosusceptible site. Treatment with sulforaphane, a dietary antioxidant, activated Nrf2 and suppressed p38-VCAM-1 signaling at the susceptible site in wild-type but not Nrf2(-/-) animals, indicating that it suppresses EC activation via Nrf2. Studies of cultured ECs revealed that Nrf2 inactivates p38 by suppressing an upstream activator MKK3/6 and by enhancing the activity of the negative regulator MKP-1. CONCLUSIONS: Nrf2 prevents ECs at the atheroprotected site from exhibiting a proinflammatory state via the suppression of p38-VCAM-1 signaling. Pharmacological activation of Nrf2 reduces EC activation at atherosusceptible sites and may provide a novel therapeutic strategy to prevent or reduce atherosclerosis.

Finally, this full study link below makes Sulforaphane even more intriguing.

http://carcin.oxfordjournals.org/cgi/content/full/27/4/811

I've been taking the BroccoMax on average 1 pil every 2 or 3 days. I never noticed a change but I'm an "extremely" poor test subject though as everyday is a good hair day. Although, I do get the occasional feeling to my scalp.

Pretty soon, I'm going to lower my dosage from the BroccoMax down to the selenium version that contains 9 mg as winter is coming up.


BTW, I was going to update this in another thread but I have "VERY" high hopes about food being the underlining problem in my hair loss. I'm sure I'm not alone. I'm thinking before thyroid and heart disease, it's actually the foods we eat that's driving this epidemic.

Since I've gone gluten free, casein free, corn free and dairy free my rosacea has virtually disappeared! Taking Betaine HCL with each meal helped as did the BroccoMax and LED light, etc. but nothing nearly as dramatic and as fast as this! This is probably the cure for Rosacea ... at least for me. Also, after taking a shower, my chest use to get pretty red. Not anymore. I can turn up the shower to very heat and my stomach stays the same color as the rest of my body like my arms. I wonder if there is a connection between having a red stomach after a hot shower and MPB. Also, I'm betting my teeth will be whiter and I don't have any bad breath when I wake up in the morning. I use to get horrible dark circles under my eyes. Those are gone ... well not completely like a teenagers but they are normal.

I have been taking my temperature though and there hasn't been much of a change if at all, so I'm still very mildly hypothryoid.

Oh, thanks for posting those studies CausticSymmetry. I'm going to print them off.

I'm surprised that Jarrow's is the only brand offering 9mg (or 30mg) of Sulforaphane Glucosinolate. I can't find any another brand that offers anything close to that.

But anyway I have been sticking to one of the Activated Seleniums a day. Allergies seem so much more in control.

The broccomax is a god-sent to mild copd patients.

hapyman - Before Jarrow came out with their Broccomax with 30 mg of Sulforaphane, there is a brand called Oncoplex used by medical professionals for the treatment of cancer, it costs about 3 times as much.

SFN but not ABG and its precursors I3C and ascorbic acid induced Nrf2 dependent gene expression at a relatively low concentration (5 mumol/l).

What's meant by 'mumol/l'? micromoles? Would such concentrations be achieved by sulforaphane-containing supplements? I think the one in IH's supplement line contains 5 mg of SFN.

Any updates on this? CS?

any comment for this one?
http://www.progressivenutracare.com/product-p/1271.htm

Looks pricey, is it worth the money?
I'm thinking to combine it with S-Acetyl Glutathoine

I already use Jarrow Formulas, Selenium Synergy, 200 mcg

Will have some updates on sulforaphane in the near future.

Regarding the Nrf2 combo, it's not too easy on the wallet. Probably the only
negative thing that can be said. Antioxidant boost is also a Nrf2 packed combo,
and much lighter on the pocket book.