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. 2021 Mar 18;70(6):1303–1316. doi: 10.2337/db20-1073

Figure 5.

Figure 5

Adiponectin suppresses TNF-α–induced MCP-1 expression and liver inflammation. A: TNF-α–induced MCP-1 expression in primary mouse hepatocytes pretreated with 50 μmol/L AdipoRon or vehicle (DMSO) for 1 h followed by 10 ng/mL TNF-α treatment for 24 h. B: Overexpression of APPL2 diminishes the inhibitory effect of adiponectin on MCP-1 expression. Primary hepatocytes overexpressing GFP control or GFP plus APPL2 were pretreated with AdipoRon (50 μmol/L) for 1 h, followed by 20 ng/mL TNF-α treatment for 6 h. C: Rapamycin treatment is sufficient to block TNF-α–induced MCP-1 expression in hepatocytes. Primary hepatocytes were pretreated with rapamycin (20 nmol/L) for 1 h, followed by TNF-α stimulation for 24 h. D: AdipoRon or rapamycin inhibits TNF-α–induced mTORC1 activation. Wild-type primary hepatocytes were pretreated with 50 μmol/L AdipoRon, 20 nmol/L rapamycin, or DMSO (vehicle) for 1 h, followed by treatment without or with 20 ng/mL TNF-α for 1 h. E: Depletion of hepatic APPL2 reduces TNF-α–stimulated mTORC1 activation in hepatocytes. APPL2-deficient hepatocytes were pretreated with 50 μmol/L AdipoRon or DMSO for 1 h, followed by 20 ng/mL TNF-α for 1 h. F: Overexpression of a dominant-negative mutant of TSC2 blocks the inhibitory effect of adiponectin on TNF-α–stimulated MCP-1 expression. Plasmids containing wild-type or dominant-negative mutant (S1345A) of TSC2 were transfected into TSC2 KO cell lines. Twenty-four hours posttransfection, cells were pretreated with 50 μmol/L AdipoRon or DMSO vehicle for 1 h, followed by 100 ng/mL TNF-α treatment. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001, p-, phosphorylated; WT, wild type.