mtorc1 activation  (MedChemExpress)

Journal: Frontiers in Pharmacology

Article Title: TLR4 modulates simvastatin’s impact on HDL cholesterol and glycemic control

doi: 10.3389/fphar.2025.1655873

Figure Lengend Snippet: Simvastatin administration during fasting activates SREBP-2 and autophagy and augments FFA-induced APOA1 expression. (A) Huh7 cells cultured under serum-depleted conditions were treated with simvastatin (1, 5, or 10 μM) for 24 h. APOA1, PLTP, and GK mRNA levels were quantified by qPCR, normalized to the geometric mean of HPRT and B2M , and expressed as fold change relative to untreated controls. (B) Simvastatin (10 μM) enhances sodium oleate (50 μM)–induced APOA1 expression in serum-depleted Huh7 cells. Gene expression was measured by qPCR and normalized to HPRT and B2M . (C) Fasting-phase simvastatin treatment upregulates hepatic Srebf2 and its target genes in vivo . A/J mice received simvastatin by gavage for 5 weeks and were sacrificed 5 h after the final dose (ZT09); hepatic mRNA levels were normalized to Hprt . (D) Expression of autophagy-related genes ( Atg7 , Atg12 , Becn1 and others) in liver following fasting-phase simvastatin treatment. (E) Pharmacologic inhibition of autophagy attenuates simvastatin-induced APOA1 expression in Huh7 cells. Serum-starved cells were treated with simvastatin in the presence or absence of 500 µM leucine (mTORC1 activator) or 50 µM leupeptin (lysosomal protease inhibitor); APOA1 mRNA was measured by qPCR and normalized to HPRT1 and B2M . Data are mean ± SEM with individual data points overlaid (n indicated in each panel). Statistical significance: p < 0.05; * p < 0.01; ** p < 0.001 (two-way ANOVA with Tukey’s post hoc test).

Article Snippet: For fasting-mimetic conditions ( ; ), cells were washed twice with 1× PBS, incubated overnight in serum-free DMEM, and then treated for 24 h with 5 μM simvastatin alone or in combination with one or more of the following: 50 μM oleic acid (MilliporeSigma; C18:1, #O1383, purity >99% by GC); 10 μM GW6471 (MedChemExpress; #HY-15372, purity 99%), a selective PPARα antagonist; 500 μM leucine (MedChemExpress; #HY-N0486, purity 98%), which suppresses autophagy initiation via mTORC1 activation; or 50 μM leupeptin (MedChemExpress; #HY-18234A, purity 99.39%), a cysteine/serine/threonine protease inhibitor that blocks autophagic flux ( ; ; ).

Techniques: Expressing, Cell Culture, Gene Expression, In Vivo, Inhibition, Protease Inhibitor

Journal: Animal Nutrition

Article Title: Propionate promotes gluconeogenesis by regulating mechanistic target of rapamycin (mTOR) pathway in calf hepatocytes

doi: 10.1016/j.aninu.2023.07.001

Figure Lengend Snippet: Role of mammalian target of rapamycin complex 1 (mTORC1) and PGC1α in propionate-mediated regulation of the expression of gluconeogenesis-related genes in calf hepatocytes. (A–C). Calf hepatocytes were treated with NaP and rapamycin (100 nM). The expression levels of FBP1 (A), PCK1 (B), and G6PC (C) were detected by RT-qPCR. (D–F). Calf hepatocytes were treated with NaP and MHY1485 (2 μM). The expression levels of FBP1 (D), PCK1 (E), and G6PC (F) were detected by RT-qPCR. (G–I). Calf hepatocytes were treated with NaP and SR18292 (20 μM). The expression levels of FBP1 (G), PCK1 (H), and G6PC (I) were detected by RT-qPCR. Data were analyzed by two-way ANOVA. a, b, c Bars with a different letter mean a significant difference ( P < 0.05).

Article Snippet: Cells were maintained in RPMI 1640 basic medium containing 2% BSA and treated with different concentrations of PA (0, 100, 200, or 400 μM) and NaP (0, 1, 2.5, or 5 mM), alone or in combination, for 12 h. A 2 × 2 factorial arrangement was applied for the experiments: primary hepatocytes were treated with NaP (2.5 mM), the mTORC1 inhibitor rapamycin (100 nM) (V900930; Sigma Aldrich, MO, USA), the mTORC1 activator MHY1485 (2 μM) (S7811; Selleck, Shanghai, China), and the PGC1α inhibitor SR-18292 (20 μM) (S8528; Selleck) for 12 h to observe the effect of mTORC1 and PGC1α on the mRNA expression of gluconeogenic genes.

Techniques: Expressing, Quantitative RT-PCR