Figure 9.

AMPK activation is necessary for salidroside- (SAL-) mediated regulation on TXNIP/NLRP3 pathway. After treatment with SAL (100 mg·kg⁻¹·d⁻¹) for 8 weeks, the liver tissues were obtained from RD and HFD mice. The expression levels of TXNIP (a) were analyzed by immunoblot. ^†† P < 0.01 versus RD mice treated with vehicle; ^∗ P < 0.05 versus HFD mice treated with vehicle. Values are means ± s.e.m. (n = 3). After cultured overnight in the serum-free medium which contains normal glucose (NG, 5.5 mM), hepatocytes were incubated in the serum-free medium which contains 30 mM glucose and 100 nM insulin (HG) and treated with 10 μM SAL for 72 h. The expression levels of TXNIP (b) were detected by immunoblot. ^† P < 0.05 versus NG; ^∗ P < 0.05 versus HG. Values are means ± s.e.m. (n = 4). After cultured overnight in the serum-free medium which contains NG, hepatocytes were incubated in the HG and treated with 10 μM Compound C (Comp. C) for 30 min before coincubated with SAL (10 μM) for 72 h. Protein sample was extracted from hepatocytes or supernatant (SN). The levels of p-AMPK, p-ACC, and TXNIP and the activation of NLRP3 inflammasome were analyzed by immunoblot (c). ^∗ P < 0.05, ^∗∗ P < 0.01 versus HG plus vehicle; ^# P < 0.05, ^## P < 0.01 versus HG plus SAL. Values are means ± s.e.m. (n = 4). Hepatocytes were transfected with 20 nM AMPKα1/α2 or scrambled siRNA for 24 h, then incubated in HG and treated with 10 μM SAL for 48 h as indicated. The phosphorylation of AMPK and ACC and the activation of NLRP3 inflammasome were analyzed by immunoblot (d). ^∗ P < 0.05, ^∗∗ P < 0.01 versus scramble siRNA without SAL. Values are means ± s.e.m. (n = 4).