Reversing silenced AR signal with demethylating agents - A promising treatment option?

The Flavoring Agent Dihydrocoumarin Reverses Epigenetic Silencing and Inhibits Sirtuin Deacetylases

Sirtuins are a family of phylogenetically conserved nicotinamide adenine dinucleotide-dependent deacetylases that have a firmly established role in aging. Using a simple Saccharomyces cerevisiae yeast heterochromatic derepression assay, we tested a number of environmental chemicals to address the possibility that humans are exposed to sirtuin inhibitors. Here we show that dihydrocoumarin (DHC), a compound found in Melilotus officinalis (sweet clover) that is commonly added to food and cosmetics, disrupted heterochromatic silencing and inhibited yeast Sir2p as well as human SIRT1 deacetylase activity. DHC exposure in the human TK6 lymphoblastoid cell line also caused concentration-dependent increases in p53 acetylation and cytotoxicity. Flow cytometric analysis to detect annexin V binding to phosphatidylserine demonstrated that DHC increased apoptosis more than 3-fold over controls. Thus, DHC inhibits both yeast Sir2p and human SIRT1 deacetylases and increases p53 acetylation and apoptosis, a phenotype associated with senescence and aging. These findings demonstrate that humans are potentially exposed to epigenetic toxicants that inhibit sirtuin deacetylases.

ncbi.nlm.nih.gov/pmc/articles/PMC1315280/

Induction of Androgen Receptor by 1,25-Dihydroxyvitamin D3 and 9-cis Retinoic Acid in LNCaP Human Prostate Cancer Cells1

We have recently shown that 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] inhibits proliferation of LNCaP cells, an androgen-responsive human prostate cancer cell line. Also, 1,25-(OH)2D3 increases androgen receptor (AR) abundance and enhances cellular responses to androgen in these cells. In the current study, we have investigated the mechanism by which 1,25-(OH)2D3 regulates AR gene expression and the involvement of AR in the 1,25-(OH)2D3- and 9-cis retinoic acid (RA)-mediated growth inhibition of LNCaP cells. Northern blot analyses demonstrated that the steady-state messenger RNA (mRNA) level of AR was significantly increased by 1,25-(OH)2D3 in a dose-dependent manner. Time-course experiments revealed that the increase of AR mRNA by 1,25-(OH)2D3 exhibited delayed kinetics. In response to 1,25-(OH)2D3, AR mRNA levels were first detected to rise at 8 h and reached a maximal induction of 10-fold over the untreated control at 48 h; the effect was sustained at 72 h. Furthermore, the induction of AR mRNA by 1,25-(OH)2D3 was completely abolished by incubation of cells with cycloheximide, a protein synthesis inhibitor. 1,25-(OH)2D3 was unable to induce expression of an AR promoter-luciferase reporter. Together, these findings indicate that the stimulatory effect of 1,25-(OH)2D3 on AR gene expression is indirect. Western blot analyses showed an increase of AR protein in 1,25-(OH)2D3-treated cells. This increased expression of AR was followed by 1,25-(OH)2D3-induced inhibition of growth in LNCaP cells. Similar to 1,25-(OH)2D3, 9-cis RA also induced AR mRNA expression, and the effect of both hormones was additive. Moreover, 1,25-(OH)2D3 and 9-cis RA acted synergistically to inhibit LNCaP cell growth. These antiproliferative effects of 1,25-(OH)2D3 and 9-cis RA, alone or in combination, were blocked by the pure AR antagonist, Casodex. In conclusion, our results demonstrate that growth inhibition of LNCaP cells by 1,25-(OH)2D3 and 9-cis RA is mediated by an AR-dependent mechanism and preceded by the induction of AR gene expression. This finding, that differentiating agents such as vitamin D and A derivatives are potent inducers of AR, may have clinical implications in the treatment of prostate cancer.

endo.endojournals.org/cgi/content/abstract/140/3/1205

Sorry if this question seems a bit obvious (cognitive impairment), but is the takeaway from this paragraph that Vitamin D and Vitamin A increase androgen receptor function?

Yeah, basically.

I have few questions. How DNA methylation can answer these.

1- Accutan causes same effects as fin/saw palmetto.
2- tribulus causes the same problems as fin/saw. There is a case of 23 old boy on musclechatroom.com he has exactly the same symptoms and the same blood profile as most of us are having here, low total T ,low DHT , low vit D low FSH low LH etc.
3- I read an other case of Metformin where it caused hypogonadism just like us.

Are these all three things 5AR inhibitors? I ask this as DNA methylation theory is based on 5AR inhibition. If any of this is not 5AR inhibitor then why these people are having symptoms like us.
how DNA methylation theory describes gyno from using HRT?

  1. I donā€™t really know enough about Accutane or how it works to comment. It is a 5ar1 inhibitor as i understand.

  2. Some guy got hypogonadism from tribulus. Ok, did his symptoms persist for years afterwards? even after correcting his hormones? Did he feel no effect from testosterone stimulation? If he is simply hypogonadal then that is not the same thing. Perhaps provide a link.

  3. Iā€™ve never heard of Metformin, but my response is basically the same as above.

In regards to gyno from HRT i think that probably has more to do with estrogen sensitivity than DNA methylation. I know a lot of guys on here have gyno, a lot of steroid users get gyno and a lot of people who have never touched any sort of drug that alters their hormones get gyno. Personally i donā€™t have any symptoms of this.

The main thing that DNA methylation theory IMO explains which sets it apart from other theories, and generally are the main factors separating PFS from other forms of hypogonadism, are:

  1. Persistence of symptoms despite often short periods on and long periods of time off the drug.

  2. An ā€œinsensitivityā€ to androgenic effects, even when hormone profiles are well within the optimal range.

  1. Accutan can give a similar problem to finasteride. This is true. Although it has alot of different effects one of them is as a 5 alpha reductase inhibitor so a similar event could happen afterwards.

jcem.endojournals.org/cgi/content/abstract/80/4/1158

  1. Tribulus causing similar issues. Havenā€™t heard of this happening but say that after correction of his hormonal profile the symptoms persisted then this could have an epigenetic origin. Iā€™m not sure what at this point is though.

  2. Metformin is not something I have looked greatly into. It is used in diabetes and sexual dysfunction can be a consequence of diabetes or it can drop testosterone levels.

jcem.endojournals.org/cgi/content/abstract/90/3/1360

  1. Gyno is a natural consequence of increasing testosterone. More is converted via aromatase into estrogen. Steroid users have this problem. It is why finasteride can cause gyno - decreased DHT meaning increased testosterone meaning increased estrogen.

Another point I think worth mentioning is Androgens bind to Androgen receptors which then lead to a cascade of events - a signal.

The problem is either in the receptor not being expressed enough or in the signalling afterwards. What this means is that some parts of that cascade may be intact whilst others might be silenced. One might have some androgenic signs but not others if that makes sense. Or one might have none. I need to confirm this mechanism as a possibility as I have yet to do enough reading on the androgen receptor so if you find out this is wrong do let me know.

Going back to dutasteride although it seems true these problems happen less frequently in them after stopping i remember when it does happen to them they get hit bad. Worse than fin. I havenā€™t dug the story out but i remember it.

yes he used tribulus for few months and got the same sides as we have. 100% the same, low DHT, low Vit D etc. Yes he has been on TRT for 8 months and no difference. TRT is having no effect on his libido, ED and joint pain. here is the link. Please look at his blood report and see how much exactly it looks to us. Read his entire thread. I am not downplaying AR theory but want to add more understanding and angles.
musclechatroom.com/forum/showthread.php?t=14049&highlight=chemman

there is another case at
forum.bodybuilding.com/showthread.php?t=669673 just click pdf file and you can see in the study.

I have read a case exactly the same. the guy used clomid etc to reverse his hypogonadism but to no avail. it was on Messorx.com I will have to find this guy. It was lon ago but if you do search you can find him. TRT did not help him at all.if you look at the following thread some guys are talking about using clomid but it is of no avail again.
mindandmuscle.net/forum/index.php?showtopic=35802

about Gyno? My poing was that if problem is at ARs then why we get gyno even here people who have low TT and low E still have gyno. My suggestion is again this points to liver enzymes being screwed up.

Ok with tribulus man he complained of ED took tribulus to boost his testosterone felt good then crashed. Youā€™re right it sounds similar in nature. It sounds in a way the same thing that happens to us. He seems very sensitive to medications. Before he took anything he had ED. Maybe it was the start of hypogonadism and tribulus wasnā€™t enough to counteract this. In fact what if his testosterone was dropping leading to androgen hypersensitivity which upon giving a jolt with tribulus led to a change in gene expression and a degree of androgen resistance. He improves in most areas with testosterone implying it helps however:

I believe the androgen signal is probably not working the way it should in him which is why no matter how much he increases his T some of his side effects will persist. Obviously it may seem i am trying to tie too much into this idea but i am not sure how the liver theory would explain it. Be grateful if you gave a summary of this idea. Thanks.

Looking at gynae case i noted the following:

revealed a markedly decreased follicle-stimulating hormone (FSH), leutinising hor- mone (LH) and testosterone. FSH 0.59 IU/l (normal: 1.0ā€“7.0), LH 0.26 IU/l (normal: 1.0ā€“8.0), testoster- one 1.3 nmol/l (normal: 10ā€“50). Prolactin, oestra- diol and progesterone were within normal limits.

On closer questioning at this stage, the patient said that he had been taking a non-hormonal preparation derived from a plant called Tribulis terrestris, in the form of tablets as a steroid alternative to supplement his weight-training. On the assumption that this substance had caused the hormonal imbalance and hence gynaecomastia, he was advised to discontinue taking them. Two months later his sex-hormones were re-checked and they had improved, FSH 11 IU/l, LH 6.1 IU/l, testosterone 15nmol/l. The swelling in his left breast had also completely resolved.

Its mechanism of action is not fully clear. It is believed to have a central effect and increase secretion of LH and therefore increase testosterone levels. Alternatively, it may exert its effect by being metabolised into androgen-like products or stimulating the physiological transfor- mation of testosterone into more active dihydro- testosterone.11,12 In our patient both gonadotropins and testosterone were markedly reduced after taking this preparation in contrast to experimental studies on this product which showed enhancement of LH and testosterone.12 In the absence of signs or symptoms of pituitary insufficiency, in an otherwise healthy individual, we believe that this could either be due to the shut-down of the pituitary gonadal axis in the presence of exogenous substances or due to increased aromatisation and peripheral conver- sion into estrogens, both of which could contribute to development of gynaecomastia.

It seemed in this patient the tribulus lowered his testosterone causing the testosterone estrogen balance to weigh heavily in the direction of estrogen causing gynaecomastia. I donā€™t know why this happened but the results speak for themselves. He improved afterwards.

Maybe tribulus can lower testosterone production in a subset of people. Maybe this is what happened in the first case too although if this was his problem i would have expected a fuller recovery. Or maybe tribulus naturally boosts testosterone for a while and then stops working. Again this is a maybe.

As for the metformin and clomid case. Metformin drops testosterone and therefore DHT. It also increases SHBG. The idea of androgen hypersensitivity to low androgens crops up again as an explanation.

Thanks for your questions. Like you I want to get to the bottom of this. If you find my argument questionable do raise it. Only through a proper Socratic method will we make progress. We need to bash every theory to the ground and see if any are left standing. I have little time for pride so please bash away :slight_smile:

thanks 19. That is what I want. We should skin every theory untill we can see the reality. If you have checked musclechatroom.com ( the link I provided) there is more talk about hypogonadism resulting from tribulus. Tribulus is known to affect liver, I posted long ago her at propecia.SP and fin alson well known to affect liver ( sorry I am a bit lazy can some one google? this businness is consuming a lot of our time.).

About Gyno I think I am not making my points clear. What I am saying suppose my baseline was TT 800 but now what is happening after sides, you get gyno long before you reach your base line say at 500 or 600.Why is happening this. is aromatization is faster? but funny thing is many of us, including the guy on musclechat.room and myself etc, have low level of E2. Or maybe there are other enzymes causing gyno? I am sure testosterone is not being metabolized the way it should be. There are other enzymes (hidden) which are playing in the back ground.

Tribulus may cause hypogonadism by dropping testosterone levels. How it does this is unknown.

It is true fin can affect the liver

Finasteride also inhibits 5b-reductase [21]. 5b and 5a-reductases are involved in hepatic steroid metabolism, and thus finasteride might affect liver function

However, there is no reason to think this causes hypogonadal symptoms. If you can show me something Iā€™m open to anything.

Testosterone is metabolised in the liver. If there were a problem in metabolism wouldnā€™t one expect a high testosterone with a low LH? This would also mean estrogen would go up.

Iā€™m not sure i understand your question about gynaecosmastia. Testosterone either converts to DHT or estrogen or is metabolised. Are you suggesting it is converted into a different thing?

ergogenics.org/anabolenboek/index11en.html - probably not the most reliable source.

In schemes 1 and 2 we have seen that testosterone can be metabolized in two ways. In Scheme 1 the reductive metabolism leading to dihydrotestosterone and in Scheme 2 the oxidative metabolism to estradiol is shown. The reduction of testosterone takes place in target tissues like the prostate and the skin and of course metabolism takes place in the liver. In male a very small part (0.2%) of the testosterone is converted into estradiol. This process mainly takes place in adipose tissue and for about 20% in the testes.

The metabolisme of testosterone and dihydrotestosterone takes place for 90% in the liver. There reductases and dehydrogenases catalyse the reactions of the D4-double bond, the C3-carbonyl group and the C17-hydroxyl group. Finally the hydroxyl groups are connected to glucuronic acid or sulphate, followed by excretion with the urine [3] [4].

The reduction of the D4-double bond is not an equilibrium reaction and goes only in one direction to give a 5a and/or a 5b-steroid skeleton (see Scheme 3). The enzyme 5a-reductase is especially active in the prostate giving 5a-dihydrotestosterone. In the liver 5a- and 5b-reductases both are active and convert testosterone and other steroids into 5a- and 5bsteroid skeletons in ratios that depend on the structure of the steroid.

After reduction of the D4-double bond often a quick reduction of the C3-carbonyl group is observed. In the 5a-steroids there is a preference for reduction to the 3b-hydroxyl compounds. In the 5b-steroids mostly reduction to the 5a-hydroxyl compounds takes place.
The C3-and C17-dehydrogenases catalyse reversible reactions, thus reactions that can proceed in two directions. The C3-dehydrogenases can reduce the C3-carbonyl group to 3a- or 3b-hydroxyl groups, which both can be oxidized again to a carbonyl group. The C17-dehydrogenases oxidise the 17b-hydroxyl group to a carbonyl group, which can be reduced again to a 17b- or a 17a-hydroxyl group.
Finally a molecule of glucuronic acid is connected to the hydroxyl groups at C3, C17 or both by an enzyme called glucuronosyl transferase (UGT) [5]. About 10% of the hydroxyl groups are converted into sulfates by sulfatases. Both groups enhance the polarity of the whole molecule considerably and in this way the apolar steroids become soluble in water and can be excreted with the urine.

In Scemes 4 and 5 a number of the possible metabolic reactions of testosterone and dihydrotestosterone are depicted. The most important excretion products are 3a-hydroxy-5b-andronstane-17-one (also called etiocholanolone) and 3a-hydroxy-5a-androstane-17-one (also called androsterone). They are mostly excreted as glucuronides. These compounds are enclosed in a frame in schemes 4 and 5.

Why did Xyrem help ithappens? maybe.

ncbi.nlm.nih.gov/pubmed/19427877

Several small chain fatty acids, including butyrate, valproate, phenylbutyrate and its derivatives, inhibit several HDAC activities in the brain at a several hundred micromolar concentration. Gamma-hydroxy-butyrate (GHB), a natural compound found in the brain originating from the metabolism of GABA, is structurally related to these fatty acids. The average physiological tissue concentration of GHB in the brain is below 50 microM, but when GHB is administered or absorbed for therapeutic or recreative purposes, its concentration reaches several hundred micromolars. In the present scenario, we demonstrate that pharmacological concentrations of GHB significantly induce brain histone H3 acetylation with a heterogeneous distribution in the brain and reduce in vitro HDAC activity. The degree of HDAC inhibition was also different according to the region of the brain considered. Taking into account the multiple physiological and functional roles attributed to the modification of histone acetylation and its consequences at the level of gene expression, we propose that part of the therapeutic or toxic effects of high concentrations of GHB in the brain after therapeutic administration of the drug could be partly due to GHB-induced epigenetic factors. In addition, we hypothesize that GHB, being naturally synthesized in the cytosolic compartment of certain neurons, could penetrate into the nuclei and may reach sufficient levels that could significantly modulate histone acetylation and may participate in the epigenetic modification of gene expression.

Correct me if i am wrong but it sounds like for the most part the young guy on musclechatroom has improved from TRT. Also the fact that he was having problems before hand, seems to complicate things.

I was surprised you were able to find a connection between GHB and gene expression, i was just gonna say that people shouldnā€™t put too much stock in ithappens recovery. The reason being that he recovered quickly from any sexual sides and only had sleep issues. It would be like saying you quit propecia and all sides went away except a rash. You then put some calamine lotion on it and got better. Does that mean calamine lotion cured you of PFS?

i think that although he got better regarding his sexual effects we should note that Xyrem cured his sleep issues. The way i see it is that his epigenetic problem wasnā€™t very deep and Xyrem managed to nudge it in the right direction.

No the young man has not cured. He is having joint pain, ED low libido etc etc. He has been fighting for almost 8 months and if you read his recent posts he is having joint pains like most of us here which he blames on his arimidex use. Just like many here he has low Estradiol and he is now thinking to use some external Estradiol. I suggested him to incease his TRT, if he thinks his E is low.

hmmm, well this was his post from 4 days agoā€¦

Anyways, i donā€™t want to derail this thread, i see your point. This is all the more reason why we need to really get a lab based study going.

yes we are here to get better not to score points.

Attenuation of Ras signaling restores androgen sensitivity to hormone-refractory C4-2 prostate cancer cells

Abstract
Progression of prostate cancer to androgen-refractory disease is correlated with increased expression of growth factors and receptors capable of establishing autocrine and/or paracrine growth-stimulatory loops. Many of these growth factor receptors engage Ras as part of their normal signaling activities, raising the possibility that activation of endogenous c-Ras could be a common mechanism for prostate cancer progression. Here we demonstrate that inducible expression of a dominant negative form of Ras restores androgen sensitivity to a hormone-refractory prostate cancer cell line. We show that expression of RasN17 in the hormone-refractory C4-2 cell line enhances in vitro sensitivity to the growth-inhibitory action of the antiandrogen Casodex and inhibits anchorage-independent cell growth. Moreover, although induction of RasN17 by itself has no observable effect on the growth of C4-2 xenografts in intact male mice, it restores androgen dependence to the C4-2 xenografts so that they dramatically regress after surgical androgen ablation.

ncbi.nlm.nih.gov/pubmed/12702591?dopt=Abstract

Does anyone else feel alot worse on gluten? I know some people have and maybe this is why:

journals.lww.com/obgynsurvey/Abstract/2004/02000/Subfertility_and_Gastrointestinal_Disease_.23.aspx

I havenā€™t bought it but i caught a snippet.

Although the mechanism is unclear, androgen resistance may play a role, as suggested by higher
levels of luteinizing hormone (LH) and free testosterone and decreased levels of DHT. These effects
are probably reversible, as a gluten-free diet appears to restore fertility

ncbi.nlm.nih.gov/pubmed/64806

These data are consistent with a reversible tissue resistance to circulating plasma-testosterone in men with gluten enteropathy and subtotal villous atrophy.

Iā€™m not suggesting we have coeliac disease but could it be whatever has happened to us caused a similar resistance worsened by a gluten diet?

I feel better on gluten free, corrivop got better after 2 years of gluten free. Only problem is its expensive.