Role of Kaempferol Combined with Pioglitazone in the Alleviation of Inflammation and Modulation of Necroptosis and Apoptosis Pathways in NASH-induced Mice

Background: Nonalcoholic steatohepatitis (NASH) is one of the most serious health problems in the world. Apoptosis and necroptosis signaling pathways are highly implicated in NASH. Therefore, this study aimed to investigate the underlying molecular mechanism for the role of Kaempferol and pioglitazone either alone or their combination in modulating NASH. Methods:  Forty C57BL/6J male mice were divided into five groups: Control group: mice received a standard chow diet and a vehicle, NASH group: mice were allowed off the NASH protocol for 25 days, Kaempferol (KP) group: Mice were maintained on NASH protocol for 25 days and were given (40 mg/kg) KP daily via oral gavage, Pioglitazone (PIO) group: mice were maintained on NASH protocol for 25 days parallel with PIO (50 mg/kg) and daily via oral gavage, KP+PIO group: mice have received NASH protocol parallel with KP and PIO co-administration with the same dose. Results: Levels of glucose, insulin, HOMA IR, LDL-C, total cholesterol, and triglycerides concentrations were significantly reduced in the other treated groups, while HDL-C was significantly raised compared with the NASH group. Otherwise, gene expression of liver AMPK was significantly increased and PPAR γ, SREBP1, and pMLKL were significantly decreased in the other three treated groups. Protein expression of caspase 8 and RIPK3 showed a significant decrease and immunohistochemical expression of NF-κB, TNF-α, and IL-6 in KP, PIO, and KP+PIO relative to the NASH group. Conclusion: Treatment of NASH by kaempferol, pioglitazone, and their combination showed a significant ameliorative effect against NASH through modulation of apoptosis and necroptosis pathways.