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: Primer sequences for RT-qPCR of cattle.

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: Sequencing

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: Increased expression of key transcription factors of gluconeogenesis in calf hepatocytes treated with propionate. Calf hepatocytes were treated with indicated concentration of NaP for 12 h, and expression levels of hepatocyte nuclear factor 4 ( HNF4A ) (A), forkhead box O1 ( FOXO1 ) (B), c-AMP response binding protein ( CREB ) (C), and peroxisome proliferator-activated receptor gamma coactivator 1-alpha ( PGC1α ) (D) were detected by RT-qPCR. Calf hepatocytes (E, F), HepG2 (G, H), and LO2 (I, J) cells were treated with indicated concentration of NaP for 12 h, the indicated proteins were detected by Western blotting (E, G, I), and quantified by ImageJ (F, H, J). Data were analyzed by one-way ANOVA. a, b, c, d 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, Concentration Assay, Binding Assay, Quantitative RT-PCR, Western Blot

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

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: Regulation of mTOR activity and expression of gluconeogenesis-related genes in calf hepatocytes treated with palmitic acid (PA). Calf hepatocytes were treated with indicated concentration of PA for 12 h, the indicated proteins were detected by Western blotting (A), and quantified by ImageJ (B–E). Calf hepatocytes were treated with PA for 12 h, and expression levels of FOXO1 (F), CREB (G), PGC1α (H), FBP1 (I), FBP2 (J), PCK1 (K), PCK2 (L), G6PC (M), ACCS1 (N), SUCLG2 (O), MCEE (P), MMUT (Q), and PCCA (R) were detected by RT-qPCR. Data were analyzed by one-way ANOVA (B–E) and t -test (F–R). a, b, c, d 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: Activity Assay, Expressing, Concentration Assay, Western Blot, Quantitative RT-PCR