Establishment of AML-12 cell model of non-alcoholic steatohepatitis
Objective To analysis and evaluate the in vitro models of non-alcoholic steatohepatitis(NASH)inflam-mation induced by different proportional concentration gradients of OA/PA.Methods AML-12 cells were induced by different concentrations of OA/PA(2∶3 or 2∶1)to establish NASH model in vitro,and CCK-8 cells were induced by OA/PA(500 μmol/L∶250 μmol/L)to detect cytotoxicity or survival rate.The mRNA contents of TNF-α,IL-6 and TGF-β 1 in the cells were measured by qRT-PCR or Western blotting.Results It was concluded that after 24 hours or 48 hours of stimulation with a mixture of different concentrations of OA/PA,OA/PA were mixed in a 2∶1 ratio(500 μmol/L+250 µmol/L)in equal molar ratio,the cytotoxicity was significantly lower than that of 2∶3 of OA/PA(200 µmol/L+300 µmol/L)(P＜0.05).After 48 hours of stimulation,the expression levels of IL-6 and TNF-α mRNA and protein in the model group were significantly increased(P＜0.05).Conclusion In vitro model of NASH inflamma-tion AML-12 was successfully established with a 2∶1 ratio of OA(500 µmol/L)/PA(250 µmol/L).
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