上海中医药杂志2026，Vol.60Issue(7)：60-73,14.DOI:10.16305/j.1007-1334.2026.z20251216001
参葛方对酒精性肝病小鼠肝脏损伤的保护作用及其相关机制
Protective effect of Shenge Formula against liver injury in alcoholic liver disease in mice and its related mechanisms
摘要
Abstract
Objective To elucidate the mechanism of Shenge Formula(SGF)in the treatment of alcoholic liver disease(ALD)by up-regulating the adiponectin receptor 1(AdipoR1)-calcium/calmodulin-dependent protein kinase kinase 2(CaMKK2)-adenosine 5′-monophosphate-activated protein kinase(AMPK)signaling pathway in hepatocytes.Methods ① Network pharmacology was employed to screen the active components and potential targets of SGF for ALD treatment,followed by systematic analysis of its core therapeutic targets and related functional pathways.② Thirty specific pathogen-free(SPF)healthy male C57BL/6J mice were randomly divided into five groups(n=6 per group):control group,model group,low-dose SGF group(SGF-L,7.95 mg/g),medium-dose SGF group(SGF-M,15.9 mg/g),and high-dose SGF group(SGF-H,31.8 mg/g).Except for the control group,a mouse model of ALD(NIAAA)was established in all other groups.Concurrently with modeling,the treatment groups were administrated with corresponding drugs via gavage,while the control and model groups were given an equal volume of 0.9%sodium chloride solution for 10 consecutive days.Serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),and triglyceride(TG),as well as hepatic levels of superoxide dismutase(SOD)and malondialdehyde(MDA),were measured.The mRNA expression levels of inflammation-and fibrosis-related genes in liver tissues were detected by reverse transcription quantitative real-time polymerase chain reaction(RT-qPCR).Protein expression levels of AdipoR1,CaMKK2,and AMPK in liver tissues were determined by Western blot analysis.Hepatic pathological changes,lipid accumulation,and inflammatory cell infiltration were assessed by hematoxylin-eosin(HE)staining,Oil Red O staining,and immunohistochemistry.③ An in vitro ALD cell model was constructed by treating HepG2 cells with absolute ethanol.Cells were further treated with SGF-containing rat serum,combined with AdipoR1 siRNA(si-AdipoR1)and Dorsomorphin(an AMPK inhibitor),to verify the key targets of SGF in ALD treatment.Results ① Network pharmacology identified CaMKK2 as a putative target of SGF against ALD,which was mainly involved in the AMPK and calcium-related signaling pathways.② Compared with the model group,serum ALT,AST,and TG levels were significantly decreased in all SGF-treated groups,accompanied by down-regulated mRNA levels of hepatic pro-inflammatory factors and collagen-related factors,as well as markedly alleviated hepatic inflammation and lipid accumulation.③ In vitro stimulation of HepG2 cells with absolute ethanol promoted intracellular lipid deposition and suppressed the protein expression of AdipoR1,CaMKK2,and phosphorylated AMPK(p-AMPK).Treatment with SGF-containing serum up-regulated AdipoR1,CaMKK2,and p-AMPK expression levels and reduced lipid accumulation.Transfection with si-AdipoR1 abolished the ameliorative effect of SGF on lipid deposition and its activation of AdipoR1,CaMKK2,and AMPK.Dorsomorphin abrogated SGF-mediated improvements in lipid deposition and SGF-triggered AMPK activation.Conclusion SGF exerts therapeutic effects against ALD via activation of the AdipoR1-CaMKK2-AMPK signaling pathway.关键词
酒精性肝病/参葛方/脂质沉积/炎症/脂联素受体1/钙调蛋白依赖性蛋白激酶激酶2/腺苷酸活化蛋白激酶Key words
alcoholic liver disease/Shenge Formula/lipid accumulation/inflammation/AdipoR1/CaMKK2/AMPK引用本文复制引用
黄知礼,冯俊华,曹海霞,李茜,纪龙珊,梁晓晖,高月求,李曼..参葛方对酒精性肝病小鼠肝脏损伤的保护作用及其相关机制[J].上海中医药杂志,2026,60(7):60-73,14.基金项目
上海市中医临床重点实验室项目(20DZ2272200) （20DZ2272200）
