平贝母水提物通过激活AMPK/ACC通路和调节肠道菌群减轻小鼠非酒精性脂肪性肝病

Aqueous extract of Fritillaria ussuriensis attenuates nonalcoholic fatty liver disease in mice by activating AMPK/ACC pathway and regulating intestinal flora

目的:探索平贝母水提物(aqueous extract of Fritillaria ussuriensis,FU-AE)抗非酒精性脂肪性肝病的作用效果与机制。方法:通过网络药理学分析平贝母(Fritillaria ussuriensis Maxir.,FU)与非酒精性脂肪性肝病的关联,利用高脂饮食(high fat diet,HFD)+10%果糖饮水诱导小鼠构建非酒精性脂肪性肝病小鼠模型,给予小鼠平贝母水提物高、中、低三个剂量进行干预治疗。通过比色法检测实验小鼠血清中门冬氨酸氨基转移酶(aspartate aminotransferase,AST)、丙氨酸氨基转移酶(alanine aminotransferase,ALT)、甘油三酯(triglyceride,TG)、胆固醇(to-tal cholesterol,TC)、高密度脂蛋白胆固醇(high-density lipoprotein cholesterol,HDL-C)、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-C)水平;HE染色评估小鼠肝脏病理学变化,明确平贝母水提物抗非酒精性脂肪性肝病的效果;提取肝脏组织蛋白,Western blot检测AMP活化蛋白激酶(AMP-activated protein kinase,AMPK)/乙酰辅酶A羧化酶(acetyl-CoA carboxylase,ACC)通路相关蛋白的表达,明确平贝母抗非酒精性脂肪性肝病作用机制。运用16S rRNA测序技术探究盲肠内容物微生物组成等,揭示平贝母水提物对非酒精性脂肪性肝病小鼠肠道菌群结构的调节作用。结果:平贝母水提物(高剂量)能够改善非酒精性脂肪性肝病小鼠肝脂肪变性(P<0.05)。与模型组比较,平贝母水提物(高剂量)干预组AST、ALT、TG、TC、LDL-C水平均显著性下降(P<0.05),HDL-C水平显著性上升(P<0.05);平贝母水提物(高剂量)干预组能够显著性增加模型小鼠肝脏中磷酸化AMPKα、AMPKα和磷酸化ACC蛋白水平(P<0.05),显著性减少ACC蛋白的表达水平(P<0.05);并显著性增加潜在有益菌Faecalibaculum ro-dentium、Lactobacillus johnsonii和Akkermansia muciniphila的相对丰度(P<0.05)。结论:平贝母水提物可能通过激活AMPK/ACC通路和调节肠道菌群结构、改善小鼠非酒精性脂肪性肝病。

AIM:To explore the effect and mechanism of action of the aqueous extract of Fritillaria ussuriensis(FU-AE)against nonalcoholic fatty liver disease(NAFLD).METHODS:The association between Fritillaria ussuriensis Maxir.(FU)and NAFLD was analyzed by network pharmacology.A mouse model of NAFLD was induced in mice by high fat diet(HFD)+10%fructose drinking water,and three doses of Fritillaria ussuriensis aqueous extract were given to the mice for intervention.Colorimetric assay was used for detection of aspartate aminotransferase(AST),alanine aminotrans-ferase(ALT),triglyceride(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C),and low-density lipoprotein cholesterol(LDL-C)levels in the serum of experimental mice.Hematoxylin and eosin staining was used to as-sess the pathological and histological changes in the liver of mice and to clarify the anti-NAFLD effect of aqueous extracts of Fritillaria ussuriensis.Liver tissue proteins were extracted,and expression of proteins related to the AMP-activated pro-tein kinase(AMPK)/acetyl-CoA carboxylase(ACC)pathway was detected by Western blot to clarify the mechanism of an-ti-NAFLD action of Fritillaria ussuriensis.The microbial composition of cecum contents was explored using 16S rRNA se-quencing to reveal the modulatory effect of the aqueous extract of Fritillaria ussuriensis on the structure of intestinal flora in mice with nonalcoholic fatty liver disease.RESULTS:Aqueous extract of Fritillaria ussuriensis(high dose)ameliorated exogenous adipocyte infiltration in the liver of mice with NAFLD(P<0.05).AST,ALT,TG,TC and LDL-C levels were significantly decreased(P<0.05)and HDL-C levels were significantly increased(P<0.05)in the high-dose group.Aque-ous extract of Fritillaria ussuriensis(high dose)significantly increased expression of phosphorylated AMPKα,AMPKα,and phosphorylated ACC in the livers of the model mice(P<0.05),significantly reduced expression of ACC(P<0.05),and significantly increased the relative abundance of the potentially beneficial bacteria Faecalibaculum rodentium,Lacto-bacillus johnsonii,Akkermansia muciniphila(P<0.05).CONCLUSION:Aqueous extract of Fritillaria ussuriensis may ameliorate NAFLD in mice by activating the AMPK/ACC pathway and modulating the structure of intestinal flora.