MTOR Activates Warburg Effect From Endotoxin

Limon9

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As Dr. Peat referred to in his newsletters, it's no surprise that a cell-proliferative enzyme activates aerobic glycolysis. Oxidative energy production is halted so the cells may regress to a primitive growth-state. Inhibiting either mTOR or the glycolytic enzyme it upregulated was sufficient to prevent the metabolic derangement and pulmonary fibrosis typically seen in endotoxin challenges and/or sepsis. This study was on the worse end of linguistically-incoherent Chinese studies, so I've corrected the worse errors.

I recall that Ray recommended azithromycin for covid, and it is generally used for other respiratory infections.

Key Points
- in vitro endotoxin treatment of lung fibroblasts increased glycolysis, lactate and collagen synthesis (fibrosis), while decreasing oxidative phosphorylation.
- Treatment of cells with 2-deoxy-d-glucose (a classic glycolysis inhibitor) prevented these effects.
- Inhibition of glycolytic enzyme PFKFB3 with 3PO prevented these effects.
- Inhibition of mTOR (with rapamycin) prevented these effects.
- Pre-treatment of live mice with 3PO before endotoxin injection prevented these effects.

PI3K-Akt-mTOR/PFKFB3 pathway mediated lung fibroblast aerobic glycolysis and collagen synthesis in lipopolysaccharide-induced pulmonary fibrosis.
Lab Invest. 2020 Jun;100(6):801-811. Hu X et al.

Highlights
"Aerobic glycolysis, which is also known as the “Warburg effect”, oxidizes glucose far more quickly than oxidative phosphorylation, contributing to an increased production of lactate [7] and plays a major role in various diseases. Reportedly, aerobic glycolysis in lung tissues also plays a crucial role during pulmonary fibrosis [8–10] and LPS-induced sepsis [11]. However, whether LPS could directly induce aerobic glycolysis and collagen synthesis in lung fibroblasts remains unclear."
"Our previous study revealed that LPS induces proliferation and activation of lung fibroblasts via the phosphatidylinositol3-kinase-protein kinase B-mammalian target of rapamycin (PI3K-Akt-mTOR) pathway [12, 13]. Other studies also showed that PI3K-Akt-mTOR pathway activation was involved in the regulation of cellular aerobic glycolysis [14, 15], which is further associated with the expression and activation of the key enzyme 6-phosphofructo-2-kinase/fructose-2, 6-biphosphatase 3 (PFKFB3) [16]."
"To investigate the effects of aerobic glycolysis on LPS-induced collagen synthesis in lung fibroblasts, 2-DG, an inhibitor of aerobic glycolysis, was applied. MRC-5 cells were treated with LPS (1 μg/mL) for 48 h. We used ECAR [extracellular acidification rate] to measure lactate production, a representation of the glycolytic rate. Compared with the control group, LPS treatment led to the continuous growth of ECAR in the lung fibroblasts, indicating enhanced glycolysis of the cells. OCR [oxygen consumption rate], a surrogate for mitochondrial respiratory activity, decreased after LPS treatment, which indicated a transition from oxidative phosphorylation to aerobic glycolysis in lung fibroblasts, and these effects were inhibited by 2-DG treatment (Fig. 1a, b). Furthermore, lactate, the end product of aerobic glycolysis, also increased after LPS treatment and could be inhibited by 2-DG pretreatment (Fig. 1c). 2-DG pretreatment also prevented the LPS-induced increased expression of collagen I (Fig. 1d). These findings indicated that aerobic glycolysis could mediate LPS-induced collagen synthesis in the lung fibroblasts in vitro."
"To investigate whether PFKFB3 is essential in LPS-induced collagen synthesis in lung fibroblasts, we first used 3PO, a selective inhibitor of PFKFB3. As shown in Fig. 2a, the pretreatment with 3PO significantly decreased the PFKFB3 level. Furthermore, pretreatment with 3PO inhibited the aerobic glycolysis in the lung fibroblasts as indicated by the lower levels of ECAR, higher levels of OCR, and less production of lactate (Fig. 2b–d), as well as inhibited lung fibroblasts collagen synthesis (Fig. 2e)."
"As shown in Fig. 4a, b, LPS promoted the activation of the PI3K-Akt-mTOR pathway detected by the phosphorylation of Akt (p-Akt) and mTOR (p-mTOR) as well as the expression of PFKFB3 at 6 h after LPS treatment, which could be inhibited by application of PI3K-Akt inhibitor LY294002, and mTOR inhibitor rapamycin, respectively. Futhermore, this inhibition precluded the LPS-induced aerobic glycolysis and collagen synthesis in the lung fibroblasts as indicated by the decreased levels of ECAR (Fig. 4c), elevated levels of OCR (Fig. 4d), less lactate production (Fig. 4e, f), and downregulation of the expression of collagen I (Fig. 4g, h). Therefore, we speculated that inhibition of the PI3K-Akt-mTOR pathway precludes LPS-induced PFKFB3 expression as well as aerobic glycolysis and collagen synthesis in the lung fibroblasts."
". . . mice were intraperitoneally pretreated with 3PO (70 mg/kg) followed by LPS (5 mg/kg) or the same dose of saline injection for 5 consecutive days. We found that the survival rate of each group were 100%, 100%, 75%, and 90% (control, 3PO, LPS, 3PO + LPS), respectively. To fully clarify the effect of 3PO on LPS-induced pulmonary fibrosis model, HE and Masson staining were performed at 3, 7, 14, and 28 days after LPS treatment and we found that mild pulmonary fibrosis was observed at 3, 7, and 14 days after LPS treatment, and typical pulmonary fibrosis was detected at 28 days, which could be reversed by 3PO pretreatment (Fig. S1A,B)."
". . . aerobic glycolysis, first detected by Warburg, suggested that tumor cells also initiate aerobic glycolysis [what about the other way round!] to produce lactate under aerobic conditions, which in turn, promotes the proliferation of the cells [22, 23]. Recent studies have shown that not only in tumor cells, aerobic glycolysis also exhibits and plays an important role in many diseases [24], such as sepsis [25, 26] and pulmonary fibrosis [21]. Also, LPS stimulates aerobic glycolysis in macrophages raises the level of lactate [25]."
"PFKFB3 is one of the critical glycolytic enzymes that regulate the synthesis of fructose-2, 6-bisphosphate (Fru2, 6-BP), which is the potent allosteric activator of glycolytic rate-limiting enzyme 6-phosphate fructose kinase 1 (6-phosphofructo-1-kinase, PFK-1). This enzyme converts fructose-6-phosphate (F6P) into fructose-1, 6bisphosphate (Fru-1, 6-BP), and plays a key role in glycolysis [28]. Therefore, the expression and activation states of PFKFB3 are directly related to the initiation of aerobic glycolysis. It was reported that in pulmonary fibrosis, the expression of PFKFB3 is upregulated in lung tissues [21]. However, its function has not yet been clarified in LPS-induced pulmonary fibrosis. The current data confirmed that LPS upregulates the expression of PFKFB3 in both lung fibroblasts and fibrotic lungs while inhibiting the level of PFKFB3 with the pretreatment of 3PO or PFKFB3 shRNA precludes the LPS-induced aerobic glycolysis and pulmonary fibrosis. Thus, it can be speculated that PFKFB3 plays a key role in regulating LPS-induced aerobic glycolysis and pulmonary fibrosis."
"Our previous study showed that LPS induces the proliferation and activation of lung fibroblasts through the activation of PI3K-Akt-mTOR pathway [12, 29]. As a significant signaling pathway, the activated PI3K-AktmTOR pathway plays a crucial role in the initiation of glycolysis [14], activation of cell proliferation [15, 30], as well as the occurrence of pulmonary fibrosis [15, 31]. It was reported that PFKFB3 could be upregulated by mTOR [32], and the PI3K-Akt-mTOR pathway was reported to be involved in the expression of PFKFB3 in liver [33]. This study confirmed that LPS activates the PI3K-Akt-mTOR pathway accompanied by the upregulation of PFKFB3 and aerobic glycolysis in the lung fibroblasts, which could be rescued by using PI3K-Akt-mTOR selective inhibitors, LY294002, and rapamycin, respectively."
"Taken together, the current study revealed that the PI3KAkt-mTOR/PFKFB3 pathway mediates aerobic glycolysis in the lung fibroblasts during LPS-induced pulmonary fibrosis, and intervention of the activation of the PI3K-AktmTOR/PFKFB3 pathway to regulate the metabolism of lung fibroblasts could be an effective therapeutic target for LPS-induced pulmonary fibrosis. In conclusion, this study demonstrated that LPS promotes collagen synthesis in the lung fibroblasts through aerobic glycolysis via activation of the PI3K-Akt-mTOR/ PFKFB3 pathway in LPS-induced pulmonary fibrosis."
 

Blossom

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As Dr. Peat referred to in his newsletters, it's no surprise that a cell-proliferative enzyme activates aerobic glycolysis. Oxidative energy production is halted so the cells may regress to a primitive growth-state. Inhibiting either mTOR or the glycolytic enzyme it upregulated was sufficient to prevent the metabolic derangement and pulmonary fibrosis typically seen in endotoxin challenges and/or sepsis. This study was on the worse end of linguistically-incoherent Chinese studies, so I've corrected the worse errors.

I recall that Ray recommended azithromycin for covid, and it is generally used for other respiratory infections.

Key Points
- in vitro endotoxin treatment of lung fibroblasts increased glycolysis, lactate and collagen synthesis (fibrosis), while decreasing oxidative phosphorylation.
- Treatment of cells with 2-deoxy-d-glucose (a classic glycolysis inhibitor) prevented these effects.
- Inhibition of glycolytic enzyme PFKFB3 with 3PO prevented these effects.
- Inhibition of mTOR (with rapamycin) prevented these effects.
- Pre-treatment of live mice with 3PO before endotoxin injection prevented these effects.

PI3K-Akt-mTOR/PFKFB3 pathway mediated lung fibroblast aerobic glycolysis and collagen synthesis in lipopolysaccharide-induced pulmonary fibrosis.
Lab Invest. 2020 Jun;100(6):801-811. Hu X et al.

Highlights
Great information! Thank you.
@Regina
 

EvanHinkle

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This is… everything?

My own journey with Peat’s work got me to endotoxin and the bacteria that cause it after about a year and a half of experimentation. I found charcoal made me human again, but all it did was keep the endotoxin at bay, (didn’t address the root cause).

My latest game changer has been antibiotics. I’m studying up on Rapamycin as well, (is it really any wonder that an “ineffective” antibiotic taken in a pulsed, low dose at 1mg extends lifespan? Like really folks, we look into such minutia so much of the time and it literally seems like bacterial infection leads to chronic endotoxin overload, leads to symptoms that we chase our tails trying to “fix.”

This is fantastic work, thank you for sharing. It validates everything I always suspected about myself, and had nothing but n=1 to base my personal assumptions on.
 

joaquin

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This is… everything?

My own journey with Peat’s work got me to endotoxin and the bacteria that cause it after about a year and a half of experimentation. I found charcoal made me human again, but all it did was keep the endotoxin at bay, (didn’t address the root cause).

My latest game changer has been antibiotics. I’m studying up on Rapamycin as well, (is it really any wonder that an “ineffective” antibiotic taken in a pulsed, low dose at 1mg extends lifespan? Like really folks, we look into such minutia so much of the time and it literally seems like bacterial infection leads to chronic endotoxin overload, leads to symptoms that we chase our tails trying to “fix.”

This is fantastic work, thank you for sharing. It validates everything I always suspected about myself, and had nothing but n=1 to base my personal assumptions on.
Is rapamycin available without rx?
 

EvanHinkle

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Is rapamycin available without rx?
There are a few ways of accessing it. The usual international pharmacy options are there, but you can actually do tele-health counseling with a doctor out of Texas in the US, who will prescribe it for anti-aging.
 

trance

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Hold on...Anti-Peat? Fatty acids are novel nutrient factors to regulate mTORC1 lysosomal localization and apoptosis in podocytes

"This study examined the effect of three types of FFAs, saturated, monounsaturated and polyunsaturated FFAs, on podocyte apoptosis. Palmitate, a saturated FFA, induced endoplasmic reticulum (ER) stress-dependent apoptosis in podocytes. Oleate, a monounsaturated FFA, and eicosapentaenoic acid (EPA), an ω-3 polyunsaturated FFA did not induce apoptosis; rather, they antagonized palmitate-induced apoptosis. Palmitate activated mammalian target of rapamycin (mTOR) complex 1 (mTORC1), a nutrient-sensing kinase regulating a wide range of cell biology. Furthermore, inhibition of mTORC1 activity by rapamycin or siRNA for Raptor, a component of mTORC1, ameliorated palmitate-induced ER stress and apoptosis in podocytes. Activity of mTORC1 is regulated by upstream kinases and Rag/Ragulator-dependent recruitment of mTOR onto lysosomal membranes. Palmitate activated mTORC1 by enhancing recruitment of mTOR onto lysosomal membranes, which was inhibited by co-incubation with oleate or EPA. Inhibition of mTOR translocation onto lysosomes by transfection with dominant-negative forms of Rag ameliorated palmitate-induced apoptosis. This study suggests that saturated and unsaturated FFAs have opposite effects on podocyte apoptosis by regulating mTORC1 activity via its translocation onto lysosomal membranes, and the results provide a better understanding of the pathogenesis in diabetic nephropathy and a novel role of mTORC1 in cell apoptosis."
 
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Limon9

Limon9

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Anti-Peat?
"There is a threshold effect in exposure to the harmful amino acids, and this involves a very sensitive system for detecting their concentration, apparently involving the actin filament network throughout the body. Above the threshold concentration, methionine activates mTOR. Increased mTOR shortens life-span and increases the processes of aging. Increased fat in the diet increases mTOR, adding to the effects of amino acids. Several substances block the increase of mTOR, including the citrus flavonoid nobiletin, aspirin, and progesterone."

- Ray Peat's Newsletter, Q2 2022, "Aging, Energy, Progesterone"
Our foods usually contain enough PUFA, unavoidably, to make fats matter to some extent. After about twenty years of carefully avoiding them, I'm still getting about 2% of my fat as PUFA (beef, oysters, eggs, etc.). That's why I'm making an effort to increase my sugar intake, to displace some fat.

- Ray Peat Email Exchange
 

trance

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But oleate and PUFA don't activate it. So now I wonder if MUFA is the golden middle between SFA and PUFA
 

joaquin

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Palmitate indirectly activates apoptosis. Isn't that a good thing?
 

S.Holmes

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All Day Chemist in India. They ask for a scrip but you can proceed without one. MUST pulse low doses (once a week or so). It's apparently very dangerous to do otherwise.
 

Drareg

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Really appreciate your threads on mTOR and linking them to Peats previous quotes, he seemed to be moving into this area of research before he passed.
 
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Limon9

Limon9

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Really appreciate your threads on mTOR and linking them to Peats previous quotes, he seemed to be moving into this area of research before he passed.
You're welcome. Mainstream hucksters talk mTOR a lot so many fall prey to "dogmatic contrarianism," assuming it must be secretly blameless because androgens/protein/etc. But what I've seen so far is consistent with Peat's feminism that progesterone is more important than testosterone*, women therefore being more "advanced^", etc.

[*] Progesterone Summaries. Peat, Raymond F.
[^] Generative Energy: Restoring The Wholeness Of Life. Peat, Raymond F. 122.
 

Peachy

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Thank you for this. I have a friend who was diagnosed as likely having adult-onset cystic fibrosis. Maybe Rapamycin could help.
 

Drareg

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You're welcome. Mainstream hucksters talk mTOR a lot so many fall prey to "dogmatic contrarianism," assuming it must be secretly blameless because androgens/protein/etc. But what I've seen so far is consistent with Peat's feminism that progesterone is more important than testosterone*, women therefore being more "advanced^", etc.

[*] Progesterone Summaries. Peat, Raymond F.
[^] Generative Energy: Restoring The Wholeness Of Life. Peat, Raymond F. 122.
He also pointed out women have larger brains relative to body size.
 

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