Salidroside Attenuates High-Fat Diet-Induced Nonalcoholic Fatty Liver Disease via AMPK-Dependent TXNIP/NLRP3 Pathway

Figure 8

Salidroside (SAL) protects hepatocytes against ROS overproduction. 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 the indicated periods of time (0–72 h), or treated with SAL (0.1, 1, 10 μM) and N-acetylcysteine (NAC, 5 mM) for 72 h. The ROS levels were detected using DCF-DA and MitoSOX as indicated (a–d). The Oil Red O staining (e) and lipid content analysis (f) were performed. Scale bar = 200 μm. Protein sample was extracted from hepatocytes or supernatant (SN). For assessment of insulin sensitivity in vitro, hepatocytes were treated as indicated, the medium was removed, and cells were incubated in fresh serum-free DMEM containing insulin (10 nM) for 20 min before protein samples were extracted. The phosphorylation of AMPK, ACC (g), Akt, and GSK3β (h) and the activation of NLRP3 inflammasome (j) were analyzed by immunoblot. The supernatant IL-1β concentration was measured by ELISA method (i). , versus NG; , versus HG. Values are means ± s.e.m. ((a)–(d), (f)–(h), and (j): ; (e) and (i): ).

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