基于体内实验和网络药理学探究清肝二十七味丸对非酒精性脂肪性肝炎的作用及作用机制OA

AIM:Through in vivo experiments and network pharmacology,the effects and mecha-nisms of Qinggan Ershiqiwei Wan(QGW)on non-al-coholic 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 mor-phology 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 triglyc-erides(TG),total cholesterol(TC),aspartate amino-transferase(AST)and alanine aminotransferase(ALT)were detected by colorimetry.Network phar-macology was used to screen key targets of QGW active components for the treatment of NASH through databases,and gene and pathway enrich-ment analysis was performed on the key targets.The levels of interleukin-1β(IL-1β)and IL-6 were quantitatively analyzed by enzyme-linked im-munosorbent assay(ELISA),and the MAPK signal-ing pathway was experimentally validated by West-ern blot and immunohistochemistry.RESULTS:Ani-mal 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 re-vealed that QGW significantly ameliorated the dis-ordered cell arrangement,lipid accumulation,and inflammatory infiltration in liver tissues induced by HFD.Network pharmacology results revealed the screening of 18 core components,including stylop-ine,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-no-tonipetranone,and luteolin;the 25 core targets,in-cluding 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 lev-els 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 de-creased in the HQG group(P<0.05,P<0.01).CON-CLUSION:QGW alleviates hepatic lipid accumula-tion and inflammatory infiltration in HFD-induced NASH mice,with its mechanism likely related to the inhibition of the MAPK signaling pathway.