28 replies to this topic

[Phoenicis]

Disclaimer: This post is not intended to be medical advice, seek the independant advice of a medical doctor before acting on any information. This post is for research pruposes only.

 

I wanted to start a discussion on the emerging epigenetic link between folate (possibly also other methyl-donors), methionine restriction (“MR”) and possible life-extending drugs like metformin.

 

Cabreiro F. et al. (2013) showed that metformin mediated life extension via altered methionine and folate metabolism. Further, Cabreiro F. et al. (2013) show that metformin induced MR and extended lifespan dependant on:  E.coli strain, metformin sensitivity and glucose concentration. According to Cabreiro F. et al. (2013), metformin alters methionine metabolism in gut E. coli and this in turn influences host metabolism. Storeli G. et al. (2013) described metformin as a methionine restriction mimetic; the latter will be discussed again later. See:

• Storelli G. et al. (2013). Metformin, microbes, and aging. Cell Metab. 2013 Jun 4;17(6):809-11.
• Cabreiro F. et al. (2013). Metformin retards aging in C. elegans by altering microbial folate and methionine metabolism. Cell. 2013 Mar 28;153(1):228-39.

This may seem somewhat farfetched since those studies were conducted worms; however human studies have also shown decreases in serum folate and increases in homocysteine. See:

• M. G. Wulffelé et al. (2003). Effects of short-term treatment with metformin on serum concentrations of homocysteine, folate and vitamin B12 in type 2 diabetes mellitus: a randomized, placebo-controlled trial
• Stefano Palomba et al. (2012). Effects of Metformin With or Without Supplementation With Folate on Homocysteine Levels and Vascular Endothelium of Women With Polycystic Ovary Syndrome

So the question is: how important are folate levels to life-extending intervention? Nguyen TP and Clarke CF.  (2012). conclude that pharmacological interventions that manipulate the gut microbiome (and thus folate) may be viable ways to extend lifespan .This suggests that folate may be important:

• Nguyen TP and Clarke CF.  (2012). Folate status of gut microbiome affects Caenorhabditis elegans lifespan. BMC Biol.; 10:66.

The following study puts some of these findings into the context of human microbiome health:

• Eleftheria Maratos-Flier. (2013). Metabolic Disease Puts Up a Fight: Microbes, metabolism and medications. Nature Medicine19,1218–121

 

Taking a Closer look at Methionine Restriction:

 

A recent paper by Tang et al. (2015) shed light on the epigenetics behind methionine restriction and more specifically showed that when MR is initiated, "S-Adenosyl-L-methionine (SAM)" reserves are readily used up in the biosynthesize of creatine. This results in epigenetic modifications like reduced methylation and changes to gene transcription. Tang, X. et al. (2015) seem to show that these SAM reserves aren't used up when glycine or arginine are lacking; as these two substrates are required for creatine biosynthesis. See:

• Tang X, Keenan MM, Wu J, Lin C-A, Dubois L, Thompson JW, et al. (2015) Comprehensive Profiling of Amino Acid Response Uncovers Unique Methionine-Deprived Response Dependent on Intact Creatine Biosynthesis. PLoS Genet 11(4): e1005158. doi:10.1371/journal.pgen.1005158

DNA hypomethylation was likewise noted in this study:

• Johnson JE, Johnson FB (2014). Methionine Restriction Activates the Retrograde Response and Confers Both Stress Tolerance and Lifespan Extension to Yeast, Mouse and Human Cells. PLoS ONE 9(5): e97729. doi:10.1371/journal.pone.0097729

 

Methyl Donors and their Role in MR and MR Mimetics:

While we know that adequate intake of vitamin b12, folate and other methyl donors is essential for health, we may be wise to play closer attention to how much we are consuming. Since metformin may act as a methionine restriction mimetic by altering intestinal microbiome metabolism, it could either enhance regular MR significantly, or lead to dangerous elevations in homocysteine due to Vitamin b12 and folate deficiencies.

Some people attempting MR do this by following a low protein vegan diet. See:

• McCarty, MF. et al. (2009). The low-methionine content of vegan diets may make methionine restriction feasible as a life extension strategy. Med Hypotheses. 2009 Feb;72(2):125-8.

While it is hard to make generalisations, some evidence seems to show that vegans consume high amounts of folate, but very little b12:

• Gilsing AM et al. (2010). Serum concentrations of vitamin B12 and folate in British male omnivores, vegetarians and vegans: results from a cross-sectional analysis of the EPIC-Oxford cohort study. Eur J Clin Nutr. 2010 Sep;64(9):933-9.
• Obersby D. et al. (2013). Plasma total homocysteine status of vegetarians compared with omnivores: a systematic review and meta-analysis. Br J Nutr. 2013 Mar 14;109(5):785-94.

Therefore one might conclude that vitamin b12 supplements would be essential for vegans. However the correct dosage must be titrated high enough to ensure that homocysteine remains very low, while at the same time b12 levels are not ecessively high; as to cause hyper-methylation. If anyone has more info on this, your input here would be appreciated!

 

For vegans getting enough folate in their diet, additional supplementation would seem to be unnecessary and even potentially harmful. Sticking close to recommended nutritional requirements would seem to be advisable. If folate levels are essential to MR, then this issue deserves further studies.

In the case of omnivores, it seems even harder to make generalisations since diet can vary so much. The best advice for everyone would seem to be: track your dietary intake in accordance with recommended levels and ask your doctor for blood tests.

 

Nutritional Supplements that act as Methyl Sinks:

 

Generally we know that many beneficial substances can become harmful when they cause hyper-homocysteine levels. One example would pharmacological doses of niacin (nicotinic acid); but I’m unsure whether the same applies to other forms of b3:

• Basu, T. et al. (2002).Niacin (nicotinic acid) in non-physiological doses causes hyperhomocysteineaemia in Sprague–Dawley rats. British Journal of Nutrition. Volume 87, Issue 02. pp 115-119

People have reported that supplementing folate, b12 and b6 is necessary to guard against homocysteine increases while taking higher doses of niacin. It appears that interventions like pharmacological doses of niacin should be studied more carefully to ascertain what the safest dose of methyl donors would be. This could be highly individual due to dietary intake. 

 

The takeaway:

 

Folate and possibly also b12 intake should be titrated under medical supervision in order to ensure that the dose is neither inadequate, nor excessive. Further research into the link between epigenetics and the human microbiome seems warranted. How important are folate and methioine/SAMe mediated DNA methylation changes to life extension interventions? Finally, is metformin a useful MR memetic, or are the risks of side-effects, b12/folate deficiency and high homocysteine too great? Note: only medical doctors are trained to provide valid advice on the last point!

Edited by Phoenicis, 29 July 2015 - 04:22 AM.

[Darryl]

Really looking forward to digging into this when its not midnight locally. I was totally unaware of the metformin/MR connection.

 

I'll just point out that therapeutic dose niacin seems to cause hyperhomocysteineaemia by interfering with B-6 uptake.

 

Basu, T. K., & Mann, S. (1997). Vitamin B-6 normalizes the altered sulfur amino acid status of rats fed diets containing pharmacological levels of niacin without reducing niacin's hypolipidemic effects. The Journal of nutrition, 127(1), 117-121.

 

My hypothesis is that nicotinic acid interferes the mucosal pyridoxal kinases involved in B-6 absorption. Niacin and B-6 vitamers have similar structural elements.

 

Niacin:     Pyridoxine:

 

Edited by Darryl, 29 July 2015 - 05:09 AM.

[Kevnzworld]

I've been measuring my methylation ( homocysteine ) for almost ten years. I started out at a level of 12 before supplementation. It's now 7.
Years ago I read the book The Homocysteine Revolution by Dr Kilmer McCully MD. For me it was eye opening. I began by taking folic acid without much success. I later read about and began using 5 methyl folate, and saw much better results.
I also supplement methyl B12 because I don't eat red meat.
Here is a pretty well researched overview of recent studies regarding excess homocysteine.
http://www.lifeexten...ysteine/Page-01
I've been taking Metformin for both reduced blood sugar ( glycation ) reasons, reduced cancer probability, and anti aging / longevity/ AMPK activation ( http://www.ncbi.nlm....cles/PMC209533/ )
I monitor my homocysteine levels regularly for both cardiovascular health and because of my Metformin supplementation.
I didn't know that Metformin may also be implicated in MR, something that at least for me is not easy to reduce or regulate.
I've substituted BCAA's for Whey protein to eliminate excess methionine, and am now also taking Glycine in the evening.
I'll be interested to see how this thread develops....very interesting.

[Phoenicis]

Forgot to add that I once heard (forget where) that taking 500mg metformin once every 3 weeks would be enough for longevity benefits. I remember thinking to myself that this didn't sense because meaningful AMPK activation would require more. Now I'm beginning to wonder whether the effects of metformin on the microbiome could account for those claimed benefits. Perhaps the effect that metformin has on microbial folate and methioine metabolism does not require high doses? Maybe the side-effects and homocysteine elevations are dose dependant?

Another good paper on the microbiome and aging:

• Heintz, C., & Mair, W. (2014). You are what you host: microbiome modulation of the aging process. Cell, 156(3), 408-411.

Heintz, C., & Mair, W. (2014) aseem to show that reduced levels of folate as a result of metformin lead to reductions in methioine. In addition Heintz, C., & Mair, W. (2014) also argue that different bacteria ellicit unique tanscriptional resonse; with L. plantarum driving growth via TOR modulation and potentially interfering with dietary restriction.

Edited by Phoenicis, 29 July 2015 - 04:07 PM.

[Phoenicis]

I've been measuring my methylation ( homocysteine ) for almost ten years. I started out at a level of 12 before supplementation. It's now 7.
[...]
I monitor my homocysteine levels regularly for both cardiovascular health and because of my Metformin supplementation.
 

 

Have you also been getting your b12 levels checked? I'm guessing you're taking a sublingual methyl b12 (I take 1000mcg Jarrow)? I was wondering how effective that would be for maintaining b12 while on metformin. It seems sublingual would be the way to go since intestinal absorbtion would be low while on metformin.

 

I actually did start taking metformin three days ago, I was interested in it because of its potential benefits for psoriasis:

• Glossmann, H. and Reider, N. (2013). Marriage of two “Methusalem” drugs for the treatment of psoriasis? Arguments for a pilot trial with metformin as add-on for methotrexate. Dermatoendocrinol. 2013 Apr 1; 5(2): 252–263

What I can say already is there has been a notable improvement; however my psoriasis has only been mild at worst since going vegan for the MR benefits. I'll explain why I think its been acting up later on. Limiting even one amino acid like methionine causes significant epigenetic changes, often referred to as the amino acid response (“AAR”)[1]. The ARR is particulary interesting for autoimmune conditions because it normalizes Th17 immune cells in diseases like psoriasis [2]. It is interesting to note that the mechanistic target of rapamycin (“mTOR”) pathway was discovered after observing the effects of the drug rapamycin; which is used as an immunosuppressant. [3] The mTOR pathway, which senses amino acid levels, controls growth and inhibits autophagy.[4] It is now recognized to play a central role in autoimmune conditions, obesity, cancer and aging.[4] Notably MR downregulates mTOR [5] and MRed vegan diets may also activate GCN2, as well as increasing FGF-21; possibly protecting agianst autoimmune disease.[6]

 

However, I did start vaping about a year ago and noticed that the nicotine definitely induced a relapse. The reason is probably that nicotine increases VEGF and HIF-1 alpha; common factors in the pathogenesis of both cancer and psoriasis [2].  See:

• Ma, X. et al. (2014). α5 Nicotinic acetylcholine receptor mediates nicotine-induced HIF-1α and VEGF expression in non-small cell lung cancer. Toxicol Appl Pharmacol. 2014 Jul 15;278(2):172-9.

After taking glycine regularly my condition improved; for example my inner ear which was usually always covered in itchy, flakey patches (gross right?) became smooth. Now with the metformin the few tiny persistant patches that I tried to treat with tazarotene for months are disappearing and I feel great. 

 

I really think that metformin is greatly enhancing the effects of methioine restriction; as is glycine. Besides those things I also take taurine, omega 3, 2000iu Vit D, 2 x calcium+mag+zinc, ester C 1g, 1000iu methyl b12 sublingual, 500-1000mg nicotinamide.

 

Note: I'm not encouraging others to try this without a doctor's advice and supervision!

 

References:

[1] Michael S. Kilberg et al. (2012). The Transcription Factor Network Associated With the Amino Acid Response in Mammalian Cells, American Society for Nutrition. Adv. Nutr. 3: 295–306.

[2] John Y. M. Koo et al. (2014). Mild to Moderate Psoriasis, CRC Press, Third Edition

[3] Ballou, L. and Richard Z. Mechanism of action of the immunosuppressant rapamycin. Life Sci. 1996;58(5):373-95.

[4] Peral, A. (2015). mTOR activation is a biomarker and a central pathway to autoimmune disorders, cancer, obesity, and aging. Annals of the New York Academy of Sciences

[5] Cheon Lee, B. et al. (2014). Methionine restriction extends lifespan of Drosophila melanogaster under conditions of low amino-acid status. Nature Communications 5, Article number: 3592

[6] McCarty, F. (2014). GCN2 and FGF21 are likely mediators of the protection from cancer, autoimmunity, obesity, and diabetes afforded by vegan diets. Medical HypothesesVolume 83, Issue 3, September 2014, Pages 365–371

 

 

Edited by Phoenicis, 30 July 2015 - 01:14 PM.

[pamojja]

Have you also been getting your b12 levels checked? I'm guessing you're taking a sublingual methyl b12 (I take 1000mcg Jarrow)? I was wondering how effective that would be for maintaining b12 while on metformin. It seems sublingual would be the way to go since intestinal absorbtion would be low while on metformin.

 

On an experiment with 1000 mg/d metformin on a vacation in a high-carb country with a regular B12 intake of about 1700 mg/d (largest part sublingual methyl- and adenosylcobalamin) and regular serum levels of about 1600 pg/ml it dropped to half of that after 6 weeks only.

 

Also I seem to be deficient in CoQ10, since whenever I drop it's dose below 160 mg/d (half that with ubiquinol) I get angina-like chest pain. In this experiment without dropping my CoQ10 intake these chest pain came back. Only after I had it confirmed that metformin also can decrease CoQ10 blood levels (for example here).

[Kevnzworld]

No, I have never had my B12 levels checked....I take 1000mcg sublingual daily
Both because I don't eat red meat and I supplement Metformin.
My biggest worry about Metformin is that it inhibits MTOR and thus muscle growth. I've been trying to figure out an effective way to cycle BCAA's, Glycine and Metformin.
I work out in the AM. I take the BCAA's then. I take a Metformin with lunch and dinner. glycine before bed.
I fear that my afternoon Metformin may be too close to my workout, and thus inhibiting muscle synthesis.

[Phoenicis]

The depletion of folate and b12 are quite well documented, the studies I cited in my first post just give us some context into what may really be going on. In the end of the day low b12 and folate are not something to messed with. I've been trying to find information on whether the effect is dose dependant, but its hard to find studies on that in particular since metformin is soes are usually raised over time. It hard to tell whether dosage of time is the critcal determinant.

 

Folate and b12 supplementation seem to critcal in the long run, the MR mimetic "benefits" must be controlled. We should try to figure out whether the effect is dose dependant and also what doses of folate/b12 have been tested to stabalise serum levels.

[Phoenicis]

Just to fix my earlier post:

 

 

The depletion of folate and b12 are quite well documented, the studies I cited in my first post just give us some context into what may really be going on. In the end of the day low b12 and folate are not something to messed with. I've been trying to find information on whether the effect is dose dependant, but its hard to find studies on that in particular, since metformin doses are usually raised over time. It hard to tell whether dosage, or time is the critcal determinant.

 

Folate and b12 supplementation seem to be critcal in the long run, the MR mimetic "benefits" must be controlled. We should try to figure out whether the effect is dose dependant and also what doses of folate/b12 have been tested to stabalise serum levels.

 

Basically we want to stabilise b12 and folate at the ideal level through supplementation. I already get aboug 780 micrograms via diet (195% RDA).

 

Xu, L. et al. (2013) recommend the supplementation of b12, but not folate; based on several studies including one that appeared to show that decreases in folate were not so significant when changes to BMI were considered. However Xu, L. et al. (2013) are merely putting forward the hypothesis that b12 supplementation alone could be advantageous. See:

• Xu, L. et al. (2013). Adverse effect of metformin therapy on serum vitamin B12 and folate: Short-term treatment causes disadvantages?. Medical hypotheses, 81(2), 149-151.

Reductions in serum b12 appear to be dose dependant and b12 likely needs to be supplemented in higher amounts than the current RDA to overcome deficiency:

• Liu, Qilin, et al. Vitamin B 12 Status in Metformin Treated Patients: Systematic Review. (2014): PLOS ONE 9(6): e100379. doi:10.1371/journal.pone.0100379

This study reports a whopping 1000mg (typo?) b12 on alternate days and then on a monthly basis (via injection?) to correct deficiency caused my metformin:

• Liu, K. W., Dai, L. K. and Jean, W. (2006). Metformin-related vitamin B12 deficiency. Age and ageing, 35(2), 200-201.

THis study likewise reported injections of 1000 mcg daily and then monthly:

• Callaghan, T. S., Hadden, D. R. and Tomkin, G. H. (1980). Megaloblastic anaemia due to vitamin B12 malabsorption associated with long-term metformin treatment. BMJ, 280(6225), 1214-1215.

This study seems to show that increased calcium intake can help with b12 absorbtion:

• Bauman, W. A. et al. (2000). Increased intake of calcium reverses vitamin B12 malabsorption induced by metformin. Diabetes care, 23(9), 1227-1231.

250mcg folate was used in this study to decrease homocysteine:

• Aarsand, A. K., & Carlsen, S. M. (1998). Folate administration reduces circulating homocysteine levels in NIDDM patients on long-term metformin treatment. JOURNAL OF INTERNAL MEDICINE-OXFORD-, 244, 169-174.

 

More on the Metformin's Effect on the Gut Microbiota:

 

Beta-glucan + blueberry extract (NM-504) and inulin modulatate microbiome and improve tolerability of metformin:

• Johnson, M. et al. (2015). Altering the Gut Microbiome to Improve Tolerability of Metformin. The FASEB Journalvol. 29 no. 1 Supplement 914.3

Metformin may significantly lower the risk of Clostridium difficile infection:

• Eliakim-Raz, N. et al. (2015). Predicting Clostridium difficile infection in diabetic patients and the effect of metformin therapy: a retrospective, case-control study. Eur J Clin Microbiol Infect Dis. 2015 Jun;34(6):1201-5.

Metformin increases number of Akkermansia "mucus-degrading Gram-negative bacteria" and reduces regulatory T cells and inflammation:

• Hur, Kyu Yeon, and Myung‐Shik Lee. (2015). New mechanisms of metformin action: Focusing on mitochondria and the gut. Journal of Diabetes Investigation (2015).

Adlercreutzia species are significantly elevated by metformin, in addiiton bile acid circulation and enteroendocrine hormone secretion are modulated by metformin:

• Napolitano, A. et al. (2014). Novel gut-based pharmacology of metformin in patients with type 2 diabetes mellitus. PLoS ONE 9(7): e100778. doi:10.1371/journal.pone.0100778

Lee, H. and Ko, G. (2014) showed that high fat diet ("HFD") increases firmicute phylum bacteria and decreases bacteroidetes phylum, wheras metformin increases bacteroidetes phylum. In contrast to eating control diet, HFD + metformin also increased phylum verrucomicrobia numbers (Lee, H. and Ko, G. (2014)). Normal diets with metformin increased  the phylums "rikenellaceae, ruminococcaceae, and verrucomicrobiaceae, as well as Alistipes spp., Akkermansia spp., and Clostridium spp." when compared with control (Lee, H. and Ko, G. (2014)). Interestingly, metformin increased the number of A. muciniphila in a dose dependant manner in the HFD group (Lee, H. and Ko, G. (2014)). Lee, H. and Ko, G. (2014) also note that "18 specific metabolic pathways of the gut microbiota were significantly increased by metformin alone"; noably these related to metabolic syndrome. See:

• Lee, H., & Ko, G. (2014). Effect of metformin on metabolic improvement and gut microbiota. Applied and environmental microbiology, 80(19), 5935-5943.

Also interesting improvements in inflammatory bowel disease:

• Lee, S. et al. (2015). Metformin ameliorates inflammatory bowel disease by suppression of STAT3 signaling pathway and regulation of the Th17/Treg balance (THER3P.1001). The Journal of Immunology; vol.194 (1 Supplement) 68.10.

 

Edited by Phoenicis, 02 August 2015 - 11:28 AM.

[Phoenicis]

Small correction:

 

Basically we want to stabilise b12 and folate at the ideal level through supplementation. I already get aboug 780 micrograms of folate via diet (195% RDA).

 

 

[Phoenicis]

Once again, sorry for reposting but wanted to clarify this:

Lee, H. and Ko, G. (2014) also note that "18 specific metabolic pathways of the gut microbiota were significantly increased by metformin alone"; notably these possibly  relate to DECREASED metabolic syndrome incidence. See:

• Lee, H., & Ko, G. (2014). Effect of metformin on metabolic improvement and gut microbiota. Applied and environmental microbiology, 80(19), 5935-5943.

 

Some more info on alternative administration routes for b12:

 

Sublingual b12 worked for a patient with short bowel syndrome and b12 malabsorbtion:

• Kotilea, K. et al. (2014). Successful Sublingual Cobalamin Treatment in a Child with Short-Bowel Syndrome. The Journal of Pediatric Pharmacology and Therapeutics, 19(1), 60-63.

No significant differences between oral and sublingual b12 found here:

• Sharabi, A., Cohen, E., Sulkes, J., & Garty, M. (2003). Replacement therapy for vitamin B12 deficiency: comparison between the sublingual and oral route. British journal of clinical pharmacology, 56(6), 635-638.

another positive study on sublingual b12:

• Delpre, G., Stark, P., & Niv, Y. (1999). Sublingual therapy for cobalamin deficiency as an alternative to oral and parenteral cobalamin supplementation. The Lancet, 354(9180), 740-741.

Intranasal b12 spray may be another option:

• Phillips, C. et al (2012). Non-invasive therapy for altered facial sensation following orthognathic surgery: an exploratory randomized clinical trial of intranasal vitamin B12 spray. Journal of maxillofacial trauma, 1(1), 20.

Edited by Phoenicis, 02 August 2015 - 06:03 PM.

[Kevnzworld]

My question regarding Metformin use is about its effects of MTOR inhibition and how it relates to muscle retention and synthesis ...MTOR is crucial in that regard. Older people are at risk for sarcopenia and age related muscle decline...Metformin could, depending on administration exacerbate that.
I'm attempting to cycle it with BCAA's and Creatine. I posted on another thread that creatine extended median lifespan of wild type mice by 9%. Roughly 8 years for humans.
http://link.springer...0726-011-0850-1
I take BCAA 's to promote muscle retention without methionine, which is in whey protein....
Thoughts?

[Phoenicis]

I think cycling is definitely an interesting approach. If you're take glycine before workouts it should boost GH. Metformin would likely attenuate mtor and GH release,but as you say it's likely dose dependant. Maybe do some research to find out more about metformin's half life and how long the effects last.

Creatine is good, I don't think anyone would dispute that. There is the issue that creatine biosynthesis is shut down by supplementation, but the significance of creatine biosynthesis mediated SAMe depletion in MR requires further study.

[Phoenicis]

In this study MR resulted mainly in decreased DNA histone methylation (CpGs) , including H3K4me2, H3K9me2, and H3K27me3:

• Tang X, Keenan MM, Wu J, Lin C-A, Dubois L, Thompson JW, et al. (2015) Comprehensive Profiling of Amino Acid Response Uncovers Unique Methionine-Deprived Response Dependent on Intact Creatine Biosynthesis. PLoS Genet 11(4): e1005158. doi:10.1371/journal.pgen.1005158

An interesting contrast to this study noting age related hypermethylation of H3K27me3 related to embryonic stem cells:

• Weidner, C. et al. (2014). Aging of blood can be tracked by DNA methylation changes at just three CpG sites. Genome biology, 15(2), R24.

Metformin was used to reprogram stem cells via epigenetics, wonder if the mechanisms are linked?:

• Oliveras-Ferraros, C. et al. (2010). Pharmacological mimicking of caloric restriction elicits epigenetic reprogramming of differentiated cells to stem-like self-renewal states. Rejuvenation research, 13(5), 519-526.

 

Edited by Phoenicis, 10 August 2015 - 01:34 PM.

[albedo]

I have been using metformin at small (250mg) doses for 2.5 months prior to the evening meal without noticing anything particular. I was not aware of the MR/metformin link but did about the muscle growth and possible effects on testosterone. I used moderate doses of creatine (~1000mg/d) prior and after resistance training and maybe have seen a positive effect on my total and free testosterone. I also checked B12, RBC folate, IGF-1, glucose, homocysteine (at 10 now but had to cut on niacin 500mg and R-ALA which likely took it to 16), liver and kidney functions. I might increase metformin till I see something on my A1C. Do you think 250 mg is a too small dose to be worth?

Edited by albedo, 11 August 2015 - 08:07 PM.

[Phoenicis]

I have been using metformin at small (250mg) doses for 2.5 months prior to the evening meal without noticing anything particular. I was not aware of the MR/metformin link but did about the muscle growth and possible effects on testosterone. I used moderate doses of creatine (~1000mg/d) prior and after resistance training and maybe have seen a positive effect on my total and free testosterone. I also checked B12, RBC folate, IGF-1, glucose, homocysteine (at 10 now but had to cut on niacin 500mg and R-ALA which likely took it to 16), liver and kidney functions. I might increase metformin till I see something on my A1C. Do you think 250 mg is a too small dose to be worth?

 

I'm definitely not qualifed or remotely confident enough to advise on the correct dose for you. From what I've seen the doses discussed by many life extensionists are notably lower than those used for treating diabetes.

 

It would be really useful to share more information about our dosing regimens and the effects they have on homocysteine, folate and b12 levels. How high are your folate and b12 levels and how do think they affected your homocysteine?

 

I'm currently taking 850mg twice per day; I know this is high dose, but I am seeing a noticeable improvement in my psoriasis. I wasn't that bad before, but my girlfriend did nitce some of the mole sized spots on my abdomen. Recently my girlfriend even commented on the improvement without me even bringing it up. I do have a b12 and folate test coming up soon, but not homocysteine. I'll try to get one in the near future. I am considering lowering my dose to 850 once per day, but will decide on that later.

 

There is a real possibility that metformin's effects on the gut microbiota could play a large role in its benefits. For example, Won-Jaelee and Kojihase (2014) report that administering Akkermansia muciniphila (bacteria increased by metformin) improved glucose tolerance and inflammation:

• Lee, W. J., & Hase, K. (2014). Gut microbiota-generated metabolites in animal health and disease. Nature chemical biology, 10(6), 416-424.

The question remains whether metformin can actually extend lifespan in species other than c. elegans. The mechanism for metformin's MR mechanism is described by Won-Jaelee and Kojihase (2014) as:

 

Metformin -> Altered folate metabolism -> reduced methionine

 

Won-Jaelee and Kojihase (2014) suggest that AMPK induction and SKN-1 (Nrf2) are mostly activated as a detoxication response to metformin. However it is unclear to me which of the two mechanisms lends itself more to the longevity benefits. It may well be a combination fo the two, but some studies seem to show that too much AMPK stimulation is indicative of too much toxicity:

• Slack, C. et al. (2012). Activation of AMPK by the putative dietary restriction mimetic metformin is insufficient to extend lifespan in Drosophila. PLoS ONE 7(10): e47699. doi:10.1371/journal.pone.0047699

It may well be possible that any longevity benefits obtained from metformin use are dose dependant. Given the fact that the MR mimetic benefits of metformin originate from changes to gut microbiota; one might ask whether low doses are sufficient. In addition, we might also ask to what extent AMPK activation is essential for longevity benefits; since both AMPK activation and toxicity increase in a dose dependant manner (Slack, C. et al. (2012)).

 

On the other hand this study showed that the protective effects of metformin against cancer increase with dose and/or time:

• Bodmer, M. et al. (2010). Long-term metformin use is associated with decreased risk of breast cancer. Diabetes care, 33(6), 1304-1308.

Menendez, J. et al. (2011) argue for chronic low dose metformin based on its mTOR deactivating properties and report that even low doses (250mg/day) were found to decrease the risk of "colorectal epithelial proliferation and aberrant crypt formation":

• Menendez, J. et al. (2011). Gerosuppressant metformin: less is more. Aging (Albany NY), 3(4), 348.

This study indicates that the increases in homocysteine seen in metformin treated diabetics are correlated to culmulative dose:

• Wile, D. J., & Toth, C. (2010). Association of metformin, elevated homocysteine, and methylmalonic acid levels and clinically worsened diabetic peripheral neuropathy. Diabetes care, 33(1), 156-161.

To me the culmulative measurement makes it hard to ascertain whether the dose quantity, or treatment time was the determining factor.

 

It's also difficult to say for sure whether homocysteine elevations are caused entirely due to decreased  folate. The mechanisms behind reduced folate and b12 are in fact different; while changes to the gut microbiome may explain alered folate levels, Bauman, W. A. et al. (2000) seem to show that "B12-intrinsic factor complex uptake" depends on "calcium availability" and  metformin basically interferes with this "calcium-dependent membrane action". See:

• Bauman, W. A. et al. (2000). Increased intake of calcium reverses vitamin B12 malabsorption induced by metformin. Diabetes care, 23(9), 1227-1231.

One might hypothesize that these effects on b12 could be highly dose and calcium dendant.

Edited by Phoenicis, 14 August 2015 - 01:53 PM.

[albedo]

 

I have been using metformin at small (250mg) doses for 2.5 months prior to the evening meal without noticing anything particular. I was not aware of the MR/metformin link but did about the muscle growth and possible effects on testosterone. I used moderate doses of creatine (~1000mg/d) prior and after resistance training and maybe have seen a positive effect on my total and free testosterone. I also checked B12, RBC folate, IGF-1, glucose, homocysteine (at 10 now but had to cut on niacin 500mg and R-ALA which likely took it to 16), liver and kidney functions. I might increase metformin till I see something on my A1C. Do you think 250 mg is a too small dose to be worth?

 

I'm definitely not qualifed or remotely confident enough to advise on the correct dose for you. From what I've seen the doses discussed by many life extensionists are notably lower than those used for treating diabetes.

 

It would be really useful to share more information about our dosing regimens and the effects they have on homocysteine, folate and b12 levels. How high are your folate and b12 levels and how do think they affected your homocysteine? ....

Thank you for your well researched post. I will go through the links but too late here.

 

In replay here are some of the values:

 Homocysteine.png   13.23KB   2 downloads

 

I probably literally bath in everything I need to have a moderate homocysteine even if my target would be in the 7-8 range (others say 4-8, see here). I also tested negative for the MTHFR SNP mutation. I will give a try to see how I react to say 1000mcg of l-methylfolate which is more absorbable but my RBC value indicate I am not doing that bad and in any case I do not intend to use high doses of folate (of course compared to RDA) for too long as many discussed on this Board (e.g. see here).  I might need to look elsewhere. E.g. I have a slight problem with RBC magnesium (slightly outside low range, despite asymptomatic) and also this might affect homocysteine. As I said, rather than adding supplements it was when I removed NA and R-ALA that I succeeded to bring it down. Of course should my low dose metformin messes things up (which is not the case so far) I might change. I need to give more time to my metformin experiment to see if I am making changes. As I was expecting, 2.5 months did not change fasting glucose and A1C. I fully agree we should post more experience with metformin dosage

[Phoenicis]

One thing about metformin; as opposed to resveratrol and rapamycin, it does not enhance stem cell regeration:

• Chen, T.et al. (2011). Rapamycin and other longevity‐promoting compounds enhance the generation of mouse induced pluripotent stem cells. Aging cell, 10(5), 908-911

Ampk activation appears to be one of the primary reasons for the above findings, however other more recent research appears to show that metformin can "efficiently generate new cells in the neonatal brain"; see:

• Dadwal, P. et al. (2015). Activating Endogenous Neural Precursor Cells Using Metformin Leads to Neural Repair and Functional Recovery in a Model of Childhood Brain Injury. Stem cell reports, 5(2), 166-173.

Also these findings seem to show that metofrmin protects hNSCs from AGEs via AMPK:

• Chung, M. et al. (2015). The neuroprotective role of metformin in advanced glycation end product treated human neural stem cells is AMPK-dependent. Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease, 1852(5), 720-731.

[Logic]

"...The authors concluded that the effects of pterostilbene were comparable to the experimental effects of 500 mg/kg oral metformin in the STZ-induced DM model...."

http://www.hindawi.c...cl/2013/575482/

 

"...A significant decrease in glucose and significant increase in plasma insulin levels were observed in normal and diabetic rats treated with pterostilbene. Treatment with pterostilbene resulted in a significant reduction of glycosylated hemoglobin and an increase in total hemoglobin level. The activities of the hepatic enzymes such as hexokinase was significantly increased whereas glucose-6-phosphatase, fructose-1,6-bisphosphatase were significantly decreased by the administration of pterostilbene in diabetic rats. A comparison was made between the action of pterostilbene and the antidiabetic drug--metformin."

http://www.ncbi.nlm....pubmed/16616938

[albedo]

Forgot to add that I once heard (forget where) that taking 500mg metformin once every 3 weeks would be enough for longevity benefits. I remember thinking to myself that this didn't sense because meaningful AMPK activation would require more. Now I'm beginning to wonder whether the effects of metformin on the microbiome could account for those claimed benefits. Perhaps the effect that metformin has on microbial folate and methioine metabolism does not require high doses? Maybe the side-effects and homocysteine elevations are dose dependant?......

 

It is an interesting idea. Should you find the source pointing to longevity benefits even at low doses of MF I would be interested. I think your calculation of 500 mg is close to what you obtain if you convert from mice to human using the BSA (Body Surface Area) methodology.

 

This study, for example, shows an increase in the mean life span by 8% in mice using 100 mg/kg which would give (1) an equivalent human dose of about 616 mg for a human of 76 kg:

 

Effect of metformin on life span and on the development of spontaneous mammary tumors in HER-2/neu transgenic mice.

http://www.ncbi.nlm....pubmed/16125352

 

For the BSA and human equivalent dose see:

Dose translation from animal to human studies revisited

http://www.fasebj.or...t/22/3/659.long

 

(1)

 HED.PNG   9.67KB   0 downloads

 BSA.PNG   48.15KB   0 downloads

[Kevnzworld]

Re : Metformin
" Metformin for cancer and aging prevention: is it a time to make the long story short?"
http://www.impactjou...th[]=17260#bib5

[albedo]

Very informative paper on metformin Kevnzworld! Thank you for sharing! Just before the conclusions, I liked the discussion on doses which I was referring to in my previous post. My experimentation with the low dose is continuing now for 8 months and intend to make a full blood panel in Q2 2016, reporting any finding here, fully sharing Phoenicis's opinion to have a place to exchange experience.

Edited by albedo, 02 December 2015 - 10:34 PM.

[Logic]

Some of the info here that jumped out at me:

 

• metformin alters methionine metabolism in gut E. coli and this in turn influences host metabolism.
• pharmacological interventions that manipulate the gut microbiome (and thus folate) may be viable ways to extend lifespan
• Metformin may significantly lower the risk of Clostridium difficile infection:
• Metformin increases number of Akkermansia "mucus-degrading Gram-negative bacteria" and reduces regulatory T cells and inflammation
• There is a real possibility that metformin's effects on the gut microbiota could play a large role in its benefits. For example, Won-Jaelee and Kojihase (2014) report that administering Akkermansia muciniphila (bacteria increased by metformin) improved glucose tolerance and inflammation:

It seems to me that getting rid of bad bacteria or their metabolites is how metformin works?
So research into other means to control the microbiome is in order here:

 

I'll start with a local search on E. coli:

 

Manganese fights e. coli toxins

http://www.longecity...-e-coli-toxins/

 

"D MANNOSE". You can research this because the information is easily available on the net. What it is very effect at is preventing and even curing bladder infections caused by E coli.

http://www.longecity...ndpost&p=289852

 

The numbers of E. coli O157:H7, L. monocytogenes, and Salmonella Typhimurium declined by 1.08, 1.24, and 1.33 log CFU/g, respectively, in raw ground beef treated with 1% Pycnogenol after 9 days of refrigerated storage. ActiVin (1%) and oleoresin rosemary (1%) resulted in an approximately 1-log CFU/g reduction in the populations of all three pathogens after 9 days.

 

ActiVin, Pycnogenol, and Herbalox retarded the formation of TBARS by 75%, 92%, 94%, and 92%, respectively,

http://www.longecity...ndpost&p=222295

 

Pathogenic Microbes: The Major Cause of Aging

http://www.longecity...cause-of-aging/

 

forskolin can cause the pouches to kick out the bacteria, allowing antibiotics to kill them,

http://www.longecity...ndpost&p=166064

 

Infection resistance

http://www.longecity...ion-resistance/

 

The local search:

https://cse.google.c...oli &gsc.page=2

 

"We all live on bacteria shit!"  (to the tune of Yellow Submarine)    
 

 

[Logic]

A Review of Pterostilbene Antioxidant Activity and Disease Modification
...Zhang and colleagues have demonstrated that pterostilbene treatment induces autophagy in oxLDL-stimulated VECs through activation of AMP-activated protein kinase (AMPK), intracellular calcium (Ca2+), and mammalian target of rapamycin (mTOR) signaling...

...The authors concluded that the effects of pterostilbene were comparable to the experimental effects of 500 mg/kg oral metformin...

...Pterostilbene was also shown to exhibit comparable and synergistic effects when compared to medications used in the treatment of human disease, specifically clofibrate, metformin, Tamoxifen, and the chemotherapy regimen FOLFOX...

http://www.hindawi.c...cl/2013/575482/

A Pterostilbene summary by yours truly:  
http://www.pterostilbene.com/

[Kevnzworld]

Re : Logic's question. " It seems to me that getting rid of bad bacteria or their metabolites is how metformin works?"
I think it's instructive to refocus n the big picture. Yes, Metformin alters vitamin B metabolism, requiring supplementation.
Those with deficiencies may experience changes in the gut biome and micro flora.
Metformin's MOA is a combination of altering glucose metabolism, gluconeogenisis and AMPK activation.
http://www.lifeexten...ne-News/Page-01

[Logic]

Re : Logic's question. " It seems to me that getting rid of bad bacteria or their metabolites is how metformin works?"
I think it's instructive to refocus n the big picture. Yes, Metformin alters vitamin B metabolism, requiring supplementation.
Those with deficiencies may experience changes in the gut biome and micro flora.
Metformin's MOA is a combination of altering glucose metabolism, gluconeogenisis and AMPK activation.
http://www.lifeexten...ne-News/Page-01

 

 

By altering how gut bacteria metabolize what we eat or decreasing the number of undesirable bacteria ..???

 

"...bowel flora, but after disturbance is removed composition of bacterial species returns to baseline ... Intestinal microbiome and energy metabolism – animal studies ... AMPK activity is increased in a germ-free mouse which leads to .... Antibiotics also improved glucose intolerance, reduced weight gain, and decreased visceral...."

http://ebook.ecog-ob...-In-Obesity.pdf

 

etc:

https://www.google.c..._sm=93&ie=UTF-8

 

I know how everyone loves 'One drug to cure us all and from the darkness save us' but perhaps this is as easy as following the FOS, Inulin, Resistant starch, probiotic advice already on site, or a combination of the 2 may be synergistic and lead to smaller reqd doses of Metformin?

Edited by Logic, 03 December 2015 - 06:57 PM.

[Kevnzworld]

Balancing gut microflora, taking probiotics will undoubtedly be beneficial. It won't mimic Metformin's MOA.

" How Metformin Functions
Metformin reduces blood glucose levels primarily by suppressing glucose formation in the liver (hepatic gluconeogenesis).59

More importantly, it activates an enzyme called AMPK (AMP-activated protein kinase) that plays an important role in insulin signaling, systemic energy balance, and the metabolism of glucose and fats.60

[Logic]

Balancing gut microflora, taking probiotics will undoubtedly be beneficial. It won't mimic Metformin's MOA.

" How Metformin Functions
Metformin reduces blood glucose levels primarily by suppressing glucose formation in the liver (hepatic gluconeogenesis).59

More importantly, it activates an enzyme called AMPK (AMP-activated protein kinase) that plays an important role in insulin signaling, systemic energy balance, and the metabolism of glucose and fats.60

 

Thx Kev.

I'll look into it more, although time is a problem atm.
I am aware of the importance of AMPK, but still feel that this effect could be due, at least in part, to the influence M has on the gut flora.
Synergies are an interesting question.

You/everyone may want to look into Pterostilbene...
NB: that it seems to be the metabolites of especially Resveratrol and Pterostilbene that cause the differences/ discrepancies/arguments seen between in-vivo and in-vitro results.

The in-vitro results of R are likely to be very similar to the in-vivo results of P..?
 

[normalizing]

i have a question about folate supplementation. reading some of the studies its confusing to me, is supplementing with folate a good idea or bad idea? what if someone has a genetic problem getting any folate, supplementing is still good or bad?

Edited by hazy, 30 November 2016 - 02:06 AM.