Ercao Qinggan decoction regulates apoptosis of hepatocytes in mice with acute liver failure via the TLR4-mediated PI3K/Akt/GSK3β signal pathway

Abstract

Background: To clarify the inhibitory effect of Ercao Qinggan decoction (EQD) on acute liver failure (ALF) and its related mechanisms. Methods: HL-7702 hepatocytes were pretreated with TLR4 inhibitor CLI-095, glycogen synthase kinase 3 β (GSK3 β) inhibitor LiCl and different doses of EQD for 2 hours, and lipopolysaccharide (LPS) (10 μg/mL) for 24 hours. Cell apoptosis, TNF-α and IL-6 and GSK3β were detected by flow cytometry, immunofluorescence, quantitative polymerase chain reaction. After mice were gavaged with different concentrations of EQD for 12 days, ALF mouse models were established intraperitoneal injection of D-Gal/LPS. After 24 hours, the mice were euthanized and the liver tissue was stained with hematoxylin and eosin. Liver cell apoptosis, the serum levels of aspartate aminotransferase, alanine aminotransferase, TNF-α and IL-β were detected by terminal transferase-mediated dUTP nick end-labelling, enzyme linked immunosorbent assay, quantitative polymerase chain reaction, and Western blotting, respectively. These methods were also used to test the mRNA expression of Bax, Bcl-2 and the protein expression of GSK3β, p-Akt/Akt in livers. Results: CLI-095, LiCl, and EQD significantly inhibited apoptosis induced by LPS, the mRNA expression of IL-6, TNF-α and the nuclear translocation of GSK3β in HL-7702 hepatocytes. EQD dose-dependently inhibited hepatocyte apoptosis, the serum concentration of aspartate aminotransferase and ALT, the expression of TNF-α and IL-β, the ratio of p-GSK3β/GSK3β, p-Akt/Akt in alanine aminotransferase mice. Conclusion: EQD can inhibit hepatocyte apoptosis in ALF mice through regulating TLR4/PI3K/Akt/GSK3β signaling pathway.