Abstract:

AIM: Through in vivo experiments and network pharmacology, the effects and mechanisms of Qinggan Ershiqiwei Wan (QGW) on non-alcoholic steatohepatitis (NASH) was explored. METHODS: A NASH mouse model was established by feeding a high-fat diet (HFD) induction. The C57BL/6J mice were divided into three groups: the normal group (CON), the model group (HFD), and the QGW administration group (HFD+QGW, HQG), with 8 mice in each group. Starting from the 7th week, the HQG group was administered the drug continuously until the 14th week, and the body weight-related indices of each group of mice were monitored. At the end of the 14th week, the mice were dissected to observe the appearance and morphology of the liver. Pathological morphology of the liver tissue was observed using H&E staining and Oil red O staining methods. The serum levels of triglycerides (TG), total cholesterol (TC), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were detected by colorimetry. Network pharmacology was used to screen key targets of QGW active components for the treatment of NASH through databases, and gene and pathway enrichment analysis was performed on the key targets. The levels of interleukin-1β (IL-1β) and IL-6 were quantitatively analyzed by enzyme-linked immunosorbent assay (ELISA), and the MAPK signaling pathway was experimentally validated by Western blot and immunohistochemistry. RESULTS: Animal experiments demonstrated that after 8 weeks of QGW intervention in model mice, serum levels of TG, TC, ALT and AST were significantly reduced (P<0.01). H&E staining and Oil Red O staining revealed that QGW significantly ameliorated the disordered cell arrangement, lipid accumulation, and inflammatory infiltration in liver tissues induced by HFD. Network pharmacology results revealed the screening of 18 core components, including stylopine, 7β-senecioyloxyoplopa-3(14)Z, 8(10)-dien-2-one, (1R,3R,4R,5S,6S)-1-acetoxy-8-angeloxoyloxy-3, 4-epoxy-5-hydroxybisabola-7(14), 10-dien-2-one, 7β-(3-ethyl-cis-crotonoyloxy)-14-hydroxy-notonipetranone, and luteolin; the 25 core targets, including MAKP8, MAPK14, MAPK9, and IL-6, were screened, involving 20 pathways including the MAPK signaling pathway. ELISA experiments demonstrated that QGW reduced the levels of IL-1β and IL-6 in the serum of mice (P<0.01). Western blot and immunohistochemistry results indicated that, compared to the CON group, the protein levels of p-ERK/ERK, p-JNK/JNK, and p-P38/P38 were significantly increased in the HFD group (P<0.01). In comparison to the HFD group, the protein levels of p-ERK/ERK, p-JNK/JNK, and p-P38/P38 were decreased in the HQG group ( P<0.05, P<0.01). CONCLUSION: QGW alleviates hepatic lipid accumulation and inflammatory infiltration in HFD-induced NASH mice, with its mechanism likely related to the inhibition of the MAPK signaling pathway.

Key words: Qinggan Ershiqiwei Wan, NASH, network pharmacology, MAPK signaling pathway