146 replies to this topic

[Tom Andre F. (ex shinobi)]

Hi Guys,

 

Finally due to some complain I prefer to open a topic here where we will speak specially about beta-lapachone, the NQO1 master gene modulator. This mechanism will be further explained here to start with:

 

> For a fast review on this topic for those who doesnt know anything about beta-lapachone: http://www.beta-lapachone.com/

 

Now to enter deeper into the topic here is a quick summary I did on another thread:

 

from this : http://journals.plos...al.pone.0047122

 

I quoted some interesting part :

 

 

Thus, it is plausible that βL may mimic the mechanisms of CR by increasing NAD+/NADH ratio. While CR leads to an increase of NAD⁺ since a low glucose level resulted from low food uptake facilitates the production of ATP by oxidizing NADH to NAD+ [31], βL could directly facilitate the production of NAD+ through the oxidation of NADH by NQO1 [8](Fig. 1A). The increased NAD⁺ can act as a signaling molecule and activate diverse proteins that play a beneficial role in the control of glucose and lipid metabolism, which is eventually deregulated during the aging process [32].

The increased NAD⁺ by βL may also be associated with the activity of AMPK (Fig. 1B)

 

in this picture, you can see mitochondria structure was better looking in the BL group than the CR group ! 

 

 

you also see BL increased some important gene within the mitochondria such as SIRT1 and SIRT3 that seems to confirm the NAD+ increase.

 

note this article is from 2012, and the korean team performed another one in 2013 and moved further:

 

http://www.ncbi.nlm....MC3944666/#sup1

 

Thus, intracellular regulation of NAD⁺ levels through NQO1 activation might be a promising therapeutic target for the protection of cisplatin-induced acute kidney injury.

 

quinone oxidoreductase 1 (NQO1) is a cytosolic antioxidant flavoprotein that catalyzes the reduction of quinones to hydroquinones by utilizing NADH as an electron donor, which consequently increases intracellular NAD⁺ levels.^{18, 19} In addition, there is evidence that NQO1 has a role in other biological activities, including anti-inflammatory processes, the scavenging of superoxide anion radicals, and the stabilization of p53 and other tumor-suppressor proteins.^{20, 21, 22, 23, 24, 25, 26}

 

^{the reference above 18 and 19:}

 

^{http://www.ncbi.nlm....pubmed/11154736}

^{http://www.ncbi.nlm....pubmed/11309386}

 

Several activators of the NQO1 enzyme have been identified, of which β-lapachone is the best known.^{27, 28}

 

 

We found that βL protects against cisplatin-induced renal dysfunction and that this effect is mediated by Sirt1 and Sirt3 through NQO1 activation

 

and indeed if you read the full study they found out BL induced again SIRT1 and SIRT3

 

Recently, numerous studies have reported that the enzymatic activation of NQO1 by βL mediates the beneficial effects on features of metabolic syndromes, including aging, obesity, hypertension, arterial restenosis, and salt-induced renal injury,^{30, 31, 33, 34} raising the possibility that intracellular NAD⁺ increase through NQO1 activation may be a potential therapeutic target for treating various diseases. In this report, we investigated whether NQO1 enzymatic activation by βL ameliorates cisplatin-induced renal changes.

 

and the most interesting part :

 

Interestingly, the major reason for the cellular change in the NAD⁺/NADH ratio by βL in WT mice is not the reduction in NADH but the increase in NAD⁺ levels (data not shown)

 

 

 

 

 

[Tom Andre F. (ex shinobi)]

 

 

James Watson mentions several NQO1 modulators on Agingsciences blog

 

Dimethyl fumarate

Taxifolin (a flavonol like Quercetin)

Resveratrol (Pterostilbene?)

Sulforaphane

Glucosinolates in Pak Choi

Sulindac

Cisplatin

Beta-Lapachone

 

(I didn't check the sources.)

 

 

Personally I made very good experience with young Pak Choi (Bok Choy), tried it twice before going to bed, about 200g, raw and very well chewed.

 

Each time I woke up very early the next morning, well-rested (uncommon for me). It's known that circadian clock and NAD+ level are closely related.

 

I intended to investigate further before I share (chance of placebo), but it's a fairly rare vegetable over here and I'm too lazy to search...

 

All these can be added to beta-lapachone regarding NQO1 but clearly they state beta-lapachone as the real one for NQO1. Also its what think the korean team that tested a lot of compound and always choose beta-lapachone. And they seems right since i never seen such impressive vivo result such the ones they got

 

 

 

Lapachol is a derivative of Vitamin K (structure-wise), so isn't it possible that beta-lapachone is metabolized to lapachol "by accident"?

The korean team should do a human blood test measuring lapachol after beta-lapachone intake (if there is any).

 

 

Ethic, I dont see why beta-lapachone should be metabolized into lapachol, so you know maybe better about this topic. Could you explain your speculation please ? I think lapachol should have some benefits on its own however

 

Also about the derivative of vitamin K, im not sure to follow you, even if they share the napthoquinone structure, I dont see why it would be a derivative ?

[normalizing]

so whats the other natural source not to bother with that tree bark as it has toxic stuff

[Ethic]

Ethic, I dont see why beta-lapachone should be metabolized into lapachol, so you know maybe better about this topic. Could you explain your speculation please ? I think lapachol should have some benefits on its own however

 

 

Also about the derivative of vitamin K, im not sure to follow you, even if they share the napthoquinone structure, I dont see why it would be a derivative ?

 

 

 

Lapachol is a metabolite of ß-Lapachone in Handroanthus impetiginosus. Seeing how human body has to handle naphthoquinones already, I was speculating about Lapachol from ß-Lapachone supplementation.

Even if ß-Lapachone is controversially disputed, it will help us to discover a related (natural?) compound with the same or better benefits, but without the risks.

Looking forward to upcoming ß-Lapachone results (Guarente L., Brenner C.?).

[Ethic]

BTW maybe you or Logic (or anyone else reading this) want to use the opportunity and establish a general NAD+ (SIRT) thread? (->Precursors, iNAMPT, eNAMPT, NQO1, AMPK, etc.)?

[normalizing]

thats what i told him about lapachol but he insists ß-Lapachone is different completely pure isolated powder used in studies and is safe. again, im just not taken fucking serious is all

[SearchHorizon]

Tom,

 

Great stuff. That Korean paper, as you pointed out elucidates the mechanism of action behind beta-lapachone. One interesting consequence of their findings is that NQO1 must up regulate NAD production as well (unlike vitamin B3s).

 

I'd be curious as to:

 

(1) how does NQO1 up regulate NAD? Note that faster NADH oxidation is not enough, because that would result in lower NADH (which was not the case).

 

(2) without beta-lapachone, how NQO1 is typically activated in a cell?  

 

(3) If NAD/NADH ratio declines with age, is it partly due to diminished NQO1 activation? In other words, does NQO1 decrease with age?

 

 

[SearchHorizon]

Moderator's Note - User has recognized errors in this post and will repost.

 

So, I have found the answer to question (2) that I asked above. As it turns out, oxidative stress is detected by/or causes NRF2 to be expressed. This is what activates NQO1. In other words, NQO1 is the result of oxidative stress, and its mechanism of action is to shift NAD/NADH ratio. This facilitates the removal of the highly reactive, toxic material.

 

The above leads to the following hypothesis: (1) if we can introduce something that is pro-oxidative in our cells AND (2) allow NRF2 to drive NQO1 to overcompensate, then we should get pro-healthful effects (through a shift in NAD/NADH ratio). In other words, we should be able to shift NAD/NADH ratio by using a pro-oxidant.

 

It appears that DHA and EPA supports the above hypothesis. As it turns out, in vitro, oxidized EPA and DHA administration  activates NRF2, which then leads to NQO1 expression.   in other words, anti-inflammatory effects of DHA and EPA is due to their peroxidation, which would be very damaging in isolation.  Thus, it appears that the positive effects of DHA and EPA (in fish oil) are largely through hormesis.

 

One consequence of the above: given that EPA and DHA is hormetic, it is probably not a good idea take large quantities of fish oil rich with EPA and DHA. It is probably a good idea to take it not too often (to give the body sufficient time to recover) (e.g., take it once a day).

Edited by YOLF, 26 May 2016 - 10:06 PM.

[SearchHorizon]

Moderator's Note - User has recognized errors in this post and will repost.

 

Oh, one more thing about NRF2, aging, and beta-lapachone. 

 

As it turns out, in both young and old people, NRF2 is activated through exercise. The activation levels in both the young and old are about the same. However, NRF2's downstream effect is attenuated in the old. (See https://www.ncbi.nlm...bmed/27109910).

 

In plain terms, exercise does not have the same healthful effect on older folks as on young people. I'd guess that the effect of supplements that directly activate NRF2 would also be attenuated in older people.

 

Of course, this begs the question - why is NRF2's downstream effects attenuated as we get old? I am not sure, but it appears that NRF2 expression is mediated by NADH. Since our NAD decreases with aging, NADH also decreases. Thus, NRF2's downstream effect would be affected by age-related decreases in NAD+. 

 

NQO1, which is triggered by NRF2, is basically a protection mechanism against quinones. It appears that quinones are "double-edged" in that, inside a cell, it will be highly reactive. NQO1 basically protects cells from it.

 

Beta-lapachone is a quinone, I believe, and that means beta-lapachone exerts its positive effects via "hormesis." If so, all precautions that are associated with administration of hormetic substances would apply to taking beta-lapachone. 

 

 

 

Edited by YOLF, 26 May 2016 - 10:03 PM.

[Tom Andre F. (ex shinobi)]

I think NQO1 is actually more important now than NAD+... Why ? Because it seems a step above. It control its level and the ratio NAD+/NADH. Its really a master regulator gene and is kind of activated by caloric restriction. Still beta-lapachone increase it better.

 

Here is a study that handle the topic perfectly. Note it was before the vivo studies that show how great benefits offer beta-lapachone:

 

http://www.ncbi.nlm....pubmed/20361926

 

NAD(P)H:quinone acceptor oxidoreductase 1 (NQO1) is a widely-distributed FAD-dependent flavoprotein that promotes obligatory 2-electron reductions of quinones, quinoneimines, nitroaromatics, and azo dyes, at rates that are comparable with NADH or NADPH. These reductions depress quinone levels and thereby minimize opportunities for generation of reactive oxygen intermediates by redox cycling, and for depletion of intracellular thiol pools. NQO1 is a highly-inducible enzyme that is regulated by the Keap1/Nrf2/ARE pathway. Evidence for the importance of the antioxidant functions of NQO1 in combating oxidative stress is provided by demonstrations that induction of NQO1 levels or their depletion (knockout, or knockdown) are associated with decreased and increased susceptibilities to oxidative stress, respectively. Furthermore, benzene genotoxicity is markedly enhanced when NQO1 activity is compromised. Not surprisingly, human polymorphisms that suppress NQO1 activities are associated with increased predisposition to disease. Recent studies have uncovered protective roles for NQO1 that apparently are unrelated to its enzymatic activities. NQO1 binds to and thereby stabilizes the important tumor suppressor p53 against proteasomal degradation. Indeed, NQO1 appears to regulate the degradative fate of other proteins. These findings suggest that NQO1 may exercise a selective "gatekeeping" role in regulating the proteasomal degradation of specific proteins, thereby broadening the cytoprotective role of NQO1 far beyond its highly effective antioxidant functions.

 

from the same study a pic summary :

 

http://www.ncbi.nlm....0038/figure/F1/

 

 

It seems to protect against all the quinones metabolites such as estrogen, and i guess androgen ones ? and even vitamin E quinones..

 

We have to express NQO1 in all tissue

 

Then another point directly from the korean team this time that I want to share:

 

They studied beta-lapachone a LOT prior to make the studies on mice and they said they found no toxic effect  in human trials (since beta-lapachone was trialed for cancer). It is an intereting point that toxicity of quinone highly depend on metabolic state of cells. In our body condition, lower oxygen and metabolic rate (low NAD+ ratio), this molecule is extremely safe. They recommand the use of this drug for very low NAD+ ratio conditions such as obesity and diabetis or aging..

 

and here he explain better what he think about the toxicity:

 

High oxidative phosphorylation in cell culture condition ----> High NAD+ -----> more increase of NAD+  by BL ---> triggers Cell death signaling.

 

Lower oxidative phosphorylation in vivo ----> low NAD+ ----> increase of NAD+  by BL ----> beneficial effects.

 

The NAD+/NADH ratio is the matter

 

The maintenance of NAD+/NADH ratio is critical. The scientist think a transient and frequency increase of the ratio may exert therapeutic effects. NMN and CoQ may exert similar effects but the difference is duration of action. Those compound is break down or metabolized shortly. bL is long-lasting as it is compared those compounds. This long-lasting property is critical for the transitent and frequent increase of NAD+/NADH ratio.

 

They fed their mices in the study using pure beta-lapachone. The company that provided them the beta-lapachone powder was merged to KT&G, a big tabacco company in Korea. Maybe someone could contact them ?

 

The scientist does not study beta-lapachone anymore but one of their members got results again using beta-lapachone and presented his work in the DAVOS 2016 conference about aging and many was excited.

 

you can see a part presenting plant extract containing beta-lapachone results (muscles.. memory etc) on mice here:

 

It seems the plant extract is chinese ginseng. But cant found any data about it elsewhere.

 

 

 

 

 

 

 

Edited by Tom Andre F. (ex shinobi), 26 May 2016 - 10:14 PM.

[Tom Andre F. (ex shinobi)]

From the paper discussed on the NR thread: http://www.nature.co...rm.2016.14.html

 

p53 stabilization has been reported to induce transcriptional activation of sestrin 1 and sestrin 2, resulting in AMPK activation88. AMPK activation leads to increased glucose and fatty acid oxidation through activation of glucose transporters (GLUTs), HIF1α and sterol regulatory element-binding proteins (SREBPs), as well as activation of mitochondrial maintenance signalling through phosphorylation and activation of the pro-mitophagic factor UNC51‑like kinase 1 (ULK1)89. In addition, AMPK phosphorylates and activates the mTORC1 inhibitor tuberous sclerosis complex 2 (TSC2), thereby further activating autophagy and inhibiting cell growth. AMPK also positively regulates PGC1α activity, which stimulates mitochondrial biogenesis, and FOXO3A activity, which leads to activation of stress response and pro-survival pathways45. Thus, it is clear that AMPK has a central role in regulating mitochondrial function both directly and indirectly (FIG. 3b). Importantly, AMPK represents a pharmacological target, with specific activators having potential beneficial effects in various age-related diseases.

 

Since we know NQO1 is a P53 stabilizator and increase AMPK, we can link to the above...

 

Beta-lapachone should help a lot actually regarding mitochondria disfunction then

[tunt01]

Overview of the NQ01 gene by James P. Watson - Anti-Aging Firewalls

 

 

[stefan_001]

Hi Tom, others,

 

Can you share some research papers that demonstrate that NQO1 is activated by β-lapachone? My understanding is that it is the other way around, so NQO1 activates β-lapachone. Because NQO1 expression is clearly higher in cancer cells β-lapachone can do its cytotoxic anti cancer work in those. This I understand is the reason that β-lapachone is selective.

 

Stefan

Edited by stefan_001, 27 May 2016 - 09:57 AM.

[Tom Andre F. (ex shinobi)]

Hi Tom, others,

 

Can you share some research papers that demonstrate that NQO1 is activated by β-lapachone? My understanding is that it is the other way around, so NQO1 activates β-lapachone. Because NQO1 expression is clearly higher in cancer cells β-lapachone can do its cytotoxic anti cancer work in those. This I understand is the reason that β-lapachone is selective.

 

Stefan

 

Hi Stefan,

 

First, Beta-lapachone is an exogenous NQO1 co-substrate and we speak about redox activation. All cancer cell line does not express NQO1 : http://www.ncbi.nlm....pubmed/19738461

 

In melanoma B16-F10 cells that do not express high NQO1 activity, both NOQ1 and METC play a critical role in beta-Lp redox activation. In contrast, in hepatocellular carcinoma HepG2 cells expressing extremely high NQO1 activity, redox activation of beta-Lp is primarily mediated by NQO1 (METC plays a minor role). These findings will contribute to our understanding of how cancer cells are selectively killed by beta-lapachone and increase our ability to devise strategies to enhance the anticancer efficacy of this potentially novel drug while minimizing its possible adverse effects on normal cells.

 

Again: The enzymatic function of NQO1 is decreased in aged tissues and enhanced by calorie restriction (as noted here for instance : http://www.pnas.org/...ent/105/7/2325)

 

About how beta-lapachone activate NQO1, here is a good study: http://www.jbc.org/content/275/8/5416

 

"β-Lapachone activates a novel apoptotic response in a number of cell lines. We demonstrate that the enzyme NAD(P)H:quinone oxidoreductase (NQO1) substantially enhances the toxicity of β-lapachone. NQO1 expression directly correlated with sensitivity to a 4-h pulse of β-lapachone in a panel of breast cancer cell lines, and the NQO1 inhibitor, dicoumarol, significantly protected NQO1-expressing cells from all aspects of β-lapachone toxicity. Stable transfection of the NQO1-deficient cell line, MDA-MB-468, with an NQO1 expression plasmid increased apoptotic responses and lethality after β-lapachone exposure. Dicoumarol blocked both the apoptotic responses and lethality. Biochemical studies suggest that reduction of β-lapachone by NQO1 leads to a futile cycling between the quinone and hydroquinone forms, with a concomitant loss of reduced NAD(P)H. In addition, the activation of a cysteine protease, which has characteristics consistent with the neutral calcium-dependent protease, calpain, is observed after β-lapachone treatment. This is the first definitive elucidation of an intracellular target for β-lapachone in tumor cells. NQO1 could be exploited for gene therapy, radiotherapy, and/or chemopreventive interventions, since the enzyme is elevated in a number of tumor types (i.e. breast and lung) and during neoplastic transformation. "

 

The capacity of βL to modulate NAD turnover is totally dependent on NQO1 as these effects were abolished in NQO1 knockout cells : http://diabetes.diab...ontent/58/4/965

 

Inspired by the fact that CR also modulates the cellular NAD⁺/NADH ratio in aged tissues, the korean studied then the ability of beta-lapachone to mimic CR in health aged mice parameters. They actually found beta-lapachone better in all biomarker tested + lifespan group

 

Let me know your thought

[stefan_001]

 

Hi Tom, others,

 

Can you share some research papers that demonstrate that NQO1 is activated by β-lapachone? My understanding is that it is the other way around, so NQO1 activates β-lapachone. Because NQO1 expression is clearly higher in cancer cells β-lapachone can do its cytotoxic anti cancer work in those.

 

Hi Stefan,

 

First, Beta-lapachone is an exogenous NQO1 co-substrate and we speak about redox activation. ...

 

In melanoma B16-F10 cells that do not express high NQO1 activity, both NOQ1 and METC play a critical role in beta-Lp redox activation. In contrast, in hepatocellular carcinoma HepG2 cells expressing extremely high NQO1 activity, redox activation of beta-Lp is primarily mediated by NQO1 (METC plays a minor role). These findings will contribute to our understanding of how cancer cells are selectively killed by beta-lapachone ..

 

About how beta-lapachone activate NQO1, here is a good study: http://www.jbc.org/content/275/8/5416

 

"β-Lapachone activates a novel apoptotic response in a number of cell lines. .... NQO1 expression directly correlated with sensitivity to a 4-h pulse of β-lapachone in a panel of breast cancer cell lines, and the NQO1 inhibitor, dicoumarol, significantly protected NQO1-expressing cells from all aspects of β-lapachone toxicity. Stable transfection of the NQO1-deficient cell line, MDA-MB-468, with an NQO1 expression plasmid increased apoptotic responses and lethality after β-lapachone exposure.... Biochemical studies suggest that reduction of β-lapachone by NQO1 leads to a futile cycling between the quinone and hydroquinone forms, with a concomitant loss of reduced NAD(P)H. ...

 

The capacity of βL to modulate NAD turnover is totally dependent on NQO1 as these effects were abolished in NQO1 knockout cells : http://diabetes.diab...ontent/58/4/965

 

Thanks for your reply, most of the above is inline with what I wrote. The way I understand this is that beta-lapachone oxidizes NADH. But it is different than the redox cycling action of NQO1 which facilitates NAD+. It seems that  beta-lapachone depletes the cell of NADH or in my thinking takes NADH out of the energy cycle making it less efficient. The article also pretty much confirms that:

"Interestingly, the food intake of βL-treated mice decreased during the first 2 weeks of treatment and then returned to the normal state (Fig. 3B) because βL-treated mice exhibited progressive weight loss even after returning to the normal state of food intake"

 

So in that sense I agree that is mimicking calorie restriction and in response to the energy crisis the cells activate defense mechanisms. But the article never says that the mouse weight stabilizes. So what happens with continuous beta-lapachone supplemetation? The mice will continue to loose weight till.......?
 

Edited by Michael, 29 July 2017 - 06:01 PM.
trim quotes

[Tom Andre F. (ex shinobi)]

Its interesting but i think it was also discussed in the above (sorry im replying using iphone). The nadh level is not deplete using beta-lapachone but its an increase in raw nad+

Mice wa fed for a long time and all health parameter was taken care. The weight is stabilized after the nad+/nadh return back to normal. Because nad+/nadh rario is depleted in diabetic, aged and obese mice. What beta-lapachone does is increase in nad+ and regulation in the ratio using nqo1. Read the first few studies in my first post i think. I bolded some part. If you was correct it would mean if i understznd that too much nad+ is bad.. And know its not possible

Second i think unlike the mice that was fed on a regular basis we can feed ourself in a transient manner as already suggested in order to turn back the ratio to normal.

[stefan_001]

Its interesting but i think it was also discussed in the above (sorry im replying using iphone). The nadh level is not deplete using beta-lapachone but its an increase in raw nad+

Mice wa fed for a long time and all health parameter was taken care. The weight is stabilized after the nad+/nadh return back to normal. Because nad+/nadh rario is depleted in diabetic, aged and obese mice. What beta-lapachone does is increase in nad+ and regulation in the ratio using nqo1. Read the first few studies in my first post i think. I bolded some part. If you was correct it would mean if i understznd that too much nad+ is bad.. And know its not possible

Second i think unlike the mice that was fed on a regular basis we can feed ourself in a transient manner as already suggested in order to turn back the ratio to normal.

 

Hi Tom, the paper is somewhat vague on that. It says:

"These results suggest that the amount and the level of NQO1 activity in aged tissues is enough for boosting the βL-mediated production of NAD⁺ and NAD⁺-associated signaling pathways in vivo."

 

There is no measurement showing this. Just the measurement showing the ratio, somewhat strange or then I overlooked it? Also it is good to note this:

 

"we checked the survival rate of the βL diet mice used in our behavioral and physiological tests (Fig. S1). They showed a 17% increase in mean survival period compared to the control mice (the βL group: 9.1±0.2 months vs the control group: 7.8±0.3 months, p<0.001 by the log rank test), whereas the CR group showed 10% (the CR group: 8.6±0.2 months, p<0.05), suggesting that βL was more effective than CR in increasing survival rate. However, compared with the normal life-span of C57BL/6J mice [25], the life-span of the control group used in our study was shorter, probably due to the effects of stress from experimental manipulations, i. e., checking of body weight, food intake and other tests. Thus, the effects of βL on life-span remains obscure and needs to be measured again with a standard protocol.

 

 

[Tom Andre F. (ex shinobi)]

 

The weight is stabilized after the nad+/nadh return back to normal. Because nad+/nadh rario is depleted in diabetic, aged and obese mice. What beta-lapachone does is increase in nad+ and regulation in the ratio using nqo1....

 
Hi Tom, the paper is somewhat vague on that. It says:
"These results suggest that the amount and the level of NQO1 activity in aged tissues is enough for boosting the βL-mediated production of NAD⁺ and NAD⁺-associated signaling pathways in vivo."
 
There is no measurement showing this. Just the measurement showing the ratio, somewhat strange or then I overlooked it? Also it is good to note this:
 
"βL diet mice ... showed a 17% increase in mean survival period compared to the control mice ... whereas the CR group showed 10% .... However, compared with the normal life-span of C57BL/6J mice [25], the life-span of the control group used in our study was shorter, probably due to the effects of stress from experimental manipulations,.... Thus, the effects of βL on life-span remains obscure and needs to be measured again with a standard protocol.

 

 
Hi Stefan,
 
Indeed but all the vivo studies mentioned that the NAD+ level was increased and activated the associated sirtuins. Im not sure they published the graph alone however, I have to look it again carefully, but the information is very clear about that at least. And you right, the control group was slightly shorter regarding lifespan, and its highly due to the manipulation done on them. Nature was very interested to publish them but didnt only due to this reason. However the CR was above the line and thats what is important since BL group lived longer. Also the mitochondria health was perfect.like the measure they monitored (cholesterol HDL, skill test etc..)
 
As for the weight, you can still can see the curve seems to really stabilized a certain level : from month 4 to 10, its very stable and they finish with exact same body weight.. to answer your previous question. Interestingly the control finish even below this group. You can see also the beta-lapachone increase theur energy in a positiv manner.
 
Another study that explain the importance of NAD+/NADH ratio : http://online.lieber...9/ars.2007.1672

Edited by Michael, 29 July 2017 - 06:05 PM.
trim quotes

[stefan_001]

 

Hi Tom, the paper is somewhat vague on [beta-lapachone increasing nad+ using nqo1]. ... There is no measurement showing this. Just the measurement showing the ratio

 
Hi Stefan,
 
Indeed but all the vivo studies mentioned that the NAD+ level was increased and activated the associated sirtuins. ...
 
As for the weight, you can still can see the curve seems to really stabilized a certain level : from month 4 to 10, its very stable and they finish with exact same body weight..  Interestingly the control finish even below this group. You can see also the beta-lapachone increase theur energy in a positiv manner.

 
I agree the bodyweight does level out and the fact that the beta-lapachone supplementation led to longer lifespan than the CR mice. It would have been interesting to see what the mice died of. We know that beta-lapachone is a potential cancer drug. Could it be that the mice simply stayed cancer free longer?
 
Wrt to the NAD+ level this section of the report makes clear that they havent measured any increase but they dont know for sure......:
Despite that indirect evidence of the effects of the βL-diet to ameliorate aging-related decline, is consistent with and support the NAD⁺/NADH ratio theory of aging, an increase in NAD⁺ was not directly measured in tissues obtained from βL-fed mice. Although the acute intravenous administration of βL(5 mg/kg) resulted in a measurable increase in NAD⁺ molecules in vivo (Fig. 1A), the administration of βL in the diet (approximately 70–80 mg/kg/day) may have induced a transient change in the NAD⁺/NADH ratio at the cellular level, which was difficult to maintain during tissue preparation. In addition, the normal NAD⁺/NADH ratio seemed to be tightly linked to a fast metabolic rate in vivo (Fig. 1C), i.e., the production and consumption of NAD⁺ are enhanced simultaneously. Further studies will be needed to determine the crucial factor in these experiments: the ratio of NAD⁺/NADH, or their turnover rate with a constant NAD⁺/NADH ratio, or both.

Edited by Michael, 29 July 2017 - 06:16 PM.
trim quote

[Tom Andre F. (ex shinobi)]

I already asked about that and the mice strain was not cancer prone. So no the lifespan study is legit (again Nature found their result very interesting, only problem was the related problem with the control group that was bit stressed and lived bit shorter..) still the team performed an outstanding work, just look at the parameter they handled.

 

About the NAD+ increase, from your quote :

 

an increase in NAD⁺ was not directly measured in tissues obtained from βL-fed mice. Although the acute intravenous administration of βL(5 mg/kg) resulted in a measurable increase in NAD⁺ molecules in vivo (Fig. 1A),

 

so its valide... and they explain then why it was hard to measure in tissue :

 

 

the administration of βL in the diet (approximately 70–80 mg/kg/day) may have induced a transient change in the NAD⁺/NADH ratio at the cellular level, which was difficult to maintain during tissue preparation.

 

And please dont forget the more recent studies done by the korean team with the professor clearly noted : (just look at his CV, just for the record : http://mbel.kaist.ac...professor.html)

 

Interestingly, the major reason for the cellular change in the NAD⁺/NADH ratio by βL in WT mice is not the reduction in NADH but the increase in NAD⁺ levels (data not shown)

 

So no more speculation with that I think. Beta lapachone does increase NAD+ level but also control the ratio.. Thats why we meet the anti aging firewall conclusion there.. They explain clearly that NR is not the solution since this ratio is not fixed.. and as important AT LEAST as NAD+ level alone. David Sinclair's professor : George Church also conduced his research in that way : using retro virus to fix this ratio as well with NAD+ level.. Really I do believe sinclair is looking for beta lapachone because he also trust this stuff. I want to see more research conduced in vivo and would like to use it myself on a transient manner.

 

 

[stefan_001]

I already asked about that and the mice strain was not cancer prone. So no the lifespan study is legit (again Nature found their result very interesting, only problem was the related problem with the control group that was bit stressed and lived bit shorter..) still the team performed an outstanding work, just look at the parameter they handled.

 

About the NAD+ increase, from your quote :

 

an increase in NAD⁺ was not directly measured in tissues obtained from βL-fed mice. Although the acute intravenous administration of βL(5 mg/kg) resulted in a measurable increase in NAD⁺ molecules in vivo (Fig. 1A),

 

so its valide... and they explain then why it was hard to measure in tissue :

 

 

the administration of βL in the diet (approximately 70–80 mg/kg/day) may have induced a transient change in the NAD⁺/NADH ratio at the cellular level, which was difficult to maintain during tissue preparation.

 

And please dont forget the more recent studies done by the korean team with the professor clearly noted : (just look at his CV, just for the record : http://mbel.kaist.ac...professor.html)

 

Interestingly, the major reason for the cellular change in the NAD⁺/NADH ratio by βL in WT mice is not the reduction in NADH but the increase in NAD⁺ levels (data not shown)

 

So no more speculation with that I think. Beta lapachone does increase NAD+ level but also control the ratio.. Thats why we meet the anti aging firewall conclusion there.. They explain clearly that NR is not the solution since this ratio is not fixed.. and as important AT LEAST as NAD+ level alone. David Sinclair's professor : George Church also conduced his research in that way : using retro virus to fix this ratio as well with NAD+ level.. Really I do believe sinclair is looking for beta lapachone because he also trust this stuff. I want to see more research conduced in vivo and would like to use it myself on a transient manner.

 

It continuous to be somewhat contradictory story. So in case of intravenous treatment they are able to measure the increase in NAD+ and in case of supplementation they don't measure an increase but suddenly refer to problems in the measurement method.....

 

I agree there may be something interesting here but as of yet the story and research is not yet crisp.

 

[Tom Andre F. (ex shinobi)]

Here they found out that NADH increased following cisplatin damage induced on hearing and that NAD+ level was well increased by beta lapachone treatment: http://www.ncbi.nlm....pubmed/24922076

 

 

Cisplatin (cis-diaminedichloroplatinum-II) is an extensively used chemotherapeutic agent, and one of its most adverse effects is ototoxicity. A number of studies have demonstrated that these effects are related to oxidative stress and DNA damage. However, the precise mechanism underlying cisplatin-associated ototoxicity is still unclear. The cofactor nicotinamide adenine dinucleotide (NAD⁺) has emerged as a key regulator of cellular energy metabolism and homeostasis. Here, we demonstrate for the first time that, in cisplatin-mediated ototoxicity, the levels and activities of SIRT1 are suppressed by the reduction of intracellular NAD⁺ levels. We provide evidence that the decrease in SIRT1 activity and expression facilitated by increasing poly(ADP-ribose) transferase (PARP)-1 activation and microRNA-34a through p53 activation aggravates cisplatin-mediated ototoxicity. Moreover, we show that the induction of cellular NAD⁺ levels using β-lapachone (β-Lap), whose intracellular target is NQO1, prevents the toxic effects of cisplatin through the regulation of PARP-1 and SIRT1 activity. These results suggest that direct modulation of cellular NAD⁺ levels by pharmacological agents could be a promising therapeutic approach for protection from cisplatin-induced ototoxicity.

 

This full study confirm the fact that the ratio is what is important once again. You can check their graph at the end of the study (external link picture)

 

I believe beta-lapachone is much more potent to control the NAD+ metabolism than NR because NR will remain only for short term.. and this seems to be validated by the chromadex studies that advice a continual use of NR to feed a pool perforated

 

Edited by Tom Andre F. (ex shinobi), 28 May 2016 - 02:18 PM.

[Tom Andre F. (ex shinobi)]

http://www.ncbi.nlm....pubmed/26173397

 

Beta-Lapachone suppresses neuroinflammation by modulating the expression of cytokines and matrix metalloproteinases in activated microglia.

 

 

Abstract

BACKGROUND:

β-Lapachone (β-LAP) is a natural naphthoquinone compound isolated from the lapacho tree (Tabebuia sp.), and it has been used for treatment of rheumatoid arthritis, infection, and cancer. In the present study, we investigated whether β-LAP has anti-inflammatory effects under in vitro and in vivo neuroinflammatory conditions.

METHODS:

The effects of β-LAP on the expression of inducible nitric oxide synthase (iNOS), cytokines, and matrix metalloproteinases (MMPs) were examined in lipopolysaccharide (LPS)-stimulated BV2 microglial cells and rat primary microglia by ELISA, reverse transcription polymerase chain reaction (RT-PCR), and Western blot analysis. Microglial activation and the expression levels of proinflammatory molecules were measured in the LPS-injected mouse brain by immunohistochemistry and RT-PCR analysis. The detailed molecular mechanism underlying the anti-inflammatory effects of β-LAP was analyzed by electrophoretic mobility shift assay, reporter gene assay, Western blot, and RT-PCR analysis.

RESULTS:

β-LAP inhibited the expression of iNOS, proinflammatory cytokines, and MMPs (MMP-3, MMP-8, MMP-9) at mRNA and protein levels in LPS-stimulated microglia. On the other hand, β-LAP upregulated the expressions of anti-inflammatory molecules such as IL-10, heme oxygenase-1 (HO-1), and the tissue inhibitor of metalloproteinase-2 (TIMP-2). The anti-inflammatory effect of β-LAP was confirmed in an LPS-induced systemic inflammation mouse model. Thus, β-LAP inhibited microglial activation and the expressions of iNOS, proinflammatory cytokines, and MMPs in the LPS-injected mouse brain. Further mechanistic studies revealed that β-LAP exerts anti-inflammatory effects by inhibiting MAPKs, PI3K/AKT, and NF-κB/AP-1 signaling pathways in LPS-stimulated microglia. β-LAP also inhibited reactive oxygen species (ROS) production by suppressing the expression and/or phosphorylation of NADPH oxidase subunit proteins, such as p47(phox) and gp91(phox). The anti-oxidant effects of β-LAP appeared to be related with the increase of HO-1 and NQO1 via the Nrf2/anti-oxidant response element (ARE) pathway and/or the PKA pathway.

CONCLUSIONS:

The strong anti-inflammatory/anti-oxidant effects of beta-lapachone may provide preventive therapeutic potential for various neuroinflammatory disorders.

 

Edited by Tom Andre F. (ex shinobi), 28 May 2016 - 02:15 PM.

[Ethic]

Hi Tom, is there any news from the korean lab members taking ß-L? Do they still take it? Any experienced Side-Effects?

[normalizing]

tom said he read that this is present in other herbs, what is the report on this??

[adamh]

It is apparently found in pao d'arco, an herb found in the rain forest as well as in bilberry. This would seem to be a cheap way of getting the benefits of it without having to buy the purified drug which always costs a lot. If its in bilberry it might be present to some degree in blueberries, a known superfood. If it also produces some weight loss, that might explode its popularity relatively soon. It might be wise to stock up before the thundering herd gets wind of it. This sounds very good and taking herbs which have been studied and used for many years sounds quite safe.

[normalizing]

adamh, where did you find out in which herbs it is present? its kind of difficult to find this. also, this whole "superfood" fiasco is some kind of a soccer mom term and i dont like it one bit. blueberries make me gain weight, amazingly superpower!

[stefan_001]

Here they found out that NADH increased following cisplatin damage induced on hearing and that NAD+ level was well increased by beta lapachone treatment: http://www.ncbi.nlm....pubmed/24922076

 

 

Cisplatin (cis-diaminedichloroplatinum-II) is an extensively used chemotherapeutic agent, and one of its most adverse effects is ototoxicity. A number of studies have demonstrated that these effects are related to oxidative stress and DNA damage. However, the precise mechanism underlying cisplatin-associated ototoxicity is still unclear. The cofactor nicotinamide adenine dinucleotide (NAD⁺) has emerged as a key regulator of cellular energy metabolism and homeostasis. Here, we demonstrate for the first time that, in cisplatin-mediated ototoxicity, the levels and activities of SIRT1 are suppressed by the reduction of intracellular NAD⁺ levels. We provide evidence that the decrease in SIRT1 activity and expression facilitated by increasing poly(ADP-ribose) transferase (PARP)-1 activation and microRNA-34a through p53 activation aggravates cisplatin-mediated ototoxicity. Moreover, we show that the induction of cellular NAD⁺ levels using β-lapachone (β-Lap), whose intracellular target is NQO1, prevents the toxic effects of cisplatin through the regulation of PARP-1 and SIRT1 activity. These results suggest that direct modulation of cellular NAD⁺ levels by pharmacological agents could be a promising therapeutic approach for protection from cisplatin-induced ototoxicity.

 

This full study confirm the fact that the ratio is what is important once again. You can check their graph at the end of the study (external link picture)

 

I believe beta-lapachone is much more potent to control the NAD+ metabolism than NR because NR will remain only for short term.. and this seems to be validated by the chromadex studies that advice a continual use of NR to feed a pool perforated

 

Hi Tom, I dont understand the ratio discussion. The only thing it tells you is that something has changed in the metabolism compared to an earlier situation. For humans it is known this ratio shifts as we age and that NAD+ declines a lot. But ones you start to thinker with the cell metabolism the ratio doesn't tell too much in my opinion. For example if the thinkering leads to doubling of both NAD+ and NADH I think we would be better off bit and the ratio is unchanged.

 

The big difference with NR is that NR is a path that injects NAD+ from external. Here we are talking about salvage path and possible redistribution. LIke I said earlier I find it interesting and lets see what more study reveals so I will stay in this discussion. Personally however it will not change my view on NR and its usefulness. NR has good track record so far and clear mechanism. 

Edited by stefan_001, 30 May 2016 - 06:03 AM.

[Tom Andre F. (ex shinobi)]

Hi Tom, is there any news from the korean lab members taking ß-L? Do they still take it? Any experienced Side-Effects?

 

Hi Ethic,

 

unfortunately I dont speak to him anymore since a few time now and do not feel confortable to disturb him again as he does not work on beta-lapachone again. Some others of his team does.

 

from what he told me he was willing to use beta lapachone for its long lasting property on a transient and frequent use. But actually to answer your question I prefer to directly quote him here because I dont have more than that answer regarding your concern:

 

"I don't have human data for the synthetic beta laphachone. Some people including me have taken the bL extract, 3 tablets per day, but lead to no side effects as bodily symptoms and normal blood measures. In my case, there is no beneficial effects because I have no health problem. My father in law suffered from cancer in the urinary bladder took it and fully recovered but he also received surgery without anti-cancer therapy in hospital. You know there is no drug for uridary bladder cancer and surgical removal is the only way. In addition, I heard that many people reported dramaticd effects but these results could not be scientifically proved. 

 

I don't study the bL anymore but sometime I will take it if sometime I need it."

 

 

Here they found out that NADH increased following cisplatin damage induced on hearing and that NAD+ level was well increased by beta lapachone treatment: http://www.ncbi.nlm....pubmed/24922076

 

 

Cisplatin (cis-diaminedichloroplatinum-II) is an extensively used chemotherapeutic agent, and one of its most adverse effects is ototoxicity. A number of studies have demonstrated that these effects are related to oxidative stress and DNA damage. However, the precise mechanism underlying cisplatin-associated ototoxicity is still unclear. The cofactor nicotinamide adenine dinucleotide (NAD⁺) has emerged as a key regulator of cellular energy metabolism and homeostasis. Here, we demonstrate for the first time that, in cisplatin-mediated ototoxicity, the levels and activities of SIRT1 are suppressed by the reduction of intracellular NAD⁺ levels. We provide evidence that the decrease in SIRT1 activity and expression facilitated by increasing poly(ADP-ribose) transferase (PARP)-1 activation and microRNA-34a through p53 activation aggravates cisplatin-mediated ototoxicity. Moreover, we show that the induction of cellular NAD⁺ levels using β-lapachone (β-Lap), whose intracellular target is NQO1, prevents the toxic effects of cisplatin through the regulation of PARP-1 and SIRT1 activity. These results suggest that direct modulation of cellular NAD⁺ levels by pharmacological agents could be a promising therapeutic approach for protection from cisplatin-induced ototoxicity.

 

This full study confirm the fact that the ratio is what is important once again. You can check their graph at the end of the study (external link picture)

 

I believe beta-lapachone is much more potent to control the NAD+ metabolism than NR because NR will remain only for short term.. and this seems to be validated by the chromadex studies that advice a continual use of NR to feed a pool perforated

 

Hi Tom, I dont understand the ratio discussion. The only thing it tells you is that something has changed in the metabolism compared to an earlier situation. For humans it is known this ratio shifts as we age and that NAD+ declines a lot. But ones you start to thinker with the cell metabolism the ratio doesn't tell too much in my opinion. For example if the thinkering leads to doubling of both NAD+ and NADH I think we would be better off bit and the ratio is unchanged.

 

The big difference with NR is that NR is a path that injects NAD+ from external. Here we are talking about salvage path and possible redistribution. LIke I said earlier I find it interesting and lets see what more study reveals so I will stay in this discussion. Personally however it will not change my view on NR and its usefulness. NR has good track record so far and clear mechanism. 

 

 

Hi Stefan, I think I already gave enough paper reference here about that, it seems clear that the ratio play a huge role on the health and it is actually also proven in vivo in numerous studies as you seen. And as you admited: aging shift this ratio.

 

I dont think NR should be seen as an ennemy for beta-lapachone (or the opposite). The only thing is that the NAD+ pool is perforated and you have to use both. As you know aging is a multi factor disease you we need to think in an holistic approach:

 

Personally my top 3 would be:

 

beta-lapachone (its long-lasting property is critical for the transitent and frequent increase of NAD+/NADH ratio.)

NR ( as a precursor for the NAD+ pool) but this time as low dose since no need for high dose (wich is actually a bad thing on the long run), small dose will be enough if beta-lapachone is used in a transitent way

Pterostilbene (the best SIRT1 activator so far)

 

here you almost control the full aging metabolism. The scientist was also interested by this combination. I would love to see someone able to test that on mice

[normalizing]

im still waiting for a good description of that beta lapachone being present in various herbs but it might take a while it seems *yawns*