马钱苷调控AMPK信号通路改善果糖饮食小鼠肝脏脂代谢紊乱的作用机制  

Effects and mechanism of loganin regulating AMPK signaling pathway to ameliorate hepatic lipid metabolism disorders in fructose-fed mice

作　　者：熊阳昆 龚名洋 罗颖熙 屈雅琴 宋莹莹 张聪 XIONG Yangkun;GONG Mingyang;LUO Yingxi;QU Yaqin;SONG Yingying;ZHANG Cong(College of Basic Medical Sciences,China Three Gorges University,Yichang 443002,China;College of Biological&Pharmaceutical Sciences,China Three Gorges University,Yichang 443002,China;School of Pharmacy,Hubei University of Chinese Medicine,Wuhan 430065,China;Hubei Key Laboratory of Tumor Microenvironment and Immunotherapy,China Three Gorges University,Yichang 443002,China)

机构地区：[1]三峡大学基础医学院,湖北宜昌443002 [2]三峡大学生物与制药学院,湖北宜昌443002 [3]湖北中医药大学药学院,湖北武汉430065 [4]肿瘤微环境与免疫治疗湖北省重点实验室,湖北宜昌443002

出　　处：《中草药》2024年第21期7325-7334,共10页Chinese Traditional and Herbal Drugs

基　　金：湖北省自然科学基金青年基金项目(2024AFB140);中药资源与中药化学湖北省重点实验室开放课题项目(KLRCCM2303);武当特色中药研究湖北省重点实验室(湖北医药学院)开放课题项目(WDCM2024001);三峡大学人才科研启动基金项目(2024RCKJ009)。

摘　　要：目的 探究马钱苷改善果糖饮食小鼠肝脏脂代谢功能紊乱的作用及其机制。方法 将40只雄性C57BL/6小鼠随机分为对照组、模型组、非诺贝特(50 mg/kg)组和马钱苷低、高剂量(25、50 mg/kg)组,每组8只。对照组小鼠正常饮水,其余各组小鼠均给予30%果糖水饲养,同时非诺贝特组和马钱苷低、高剂量组小鼠分别ig给予相应药物治疗,每天1次,连续给药8周。实验结束后收集各组小鼠血清和肝脏样本,记录各组小鼠体质量、肝质量,并计算肝脏指数;检测各组小鼠血清中天冬氨酸氨基转移酶(aspartate aminotransferase,AST)和丙氨酸氨基转移酶(alanine aminotransferase,ALT)的活力、三酰甘油(triglyceride,TG)、总胆固醇(total cholesterol,TC)、高密度脂蛋白胆固醇(high-density lipoprotein cholesterol,HDL-C)和低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-C)的水平,同时检测各组小鼠肝脏中TG、丙二醛(malondialdehyde,MDA)、总超氧化物歧化酶(total-superoxide dismutase,T-SOD)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)、IL-1β的水平;苏木素-伊红(hematoxylin-eosin staining,HE)和油红O染色观察肝脏病理学变化;通过转录组测序筛选马钱苷干预后果糖饮食小鼠肝脏中差异表达的基因,并对差异表达基因进行功能富集分析;通过分子对接分析马钱苷与单磷酸腺苷活化蛋白激酶α(adenosine monophosphate activated protein kinase α,AMPKα)蛋白的相互作用;采用Western blotting检测各组小鼠肝组织中AMPKα、磷酸化AMPKα(phosphorylation-AMPKαThr172,p-AMPKα)、固醇调节元件结合蛋白-1(sterol regulatory element binding protein-1,SREBP-1)和脂肪酸合成酶(fatty acid synthetase,FASN)的蛋白表达量;采用免疫组织化学法检测各组小鼠肝组织中p-AMPKα蛋白表达。结果 与模型组比较,马钱苷治疗组小鼠体质量、肝质量和肝脏指数显著降低(P<0.05、0.01),血清中ASObjective To investigate the effect and mechanism of loganin ameliorating hepatic lipid metabolism disorder in fructose-fed mice.Methods Male C57BL/6 mice were randomly divided into control group,model group,fenofibrate(50 mg/kg)group,and low-,high-dose loganin group(25,50 mg/kg),with eight mice in each group.The mice in the control group were given normal drinking water,and the mice in the other groups were fed 30%fructose water,while the mice in the fenofibrate group and the low-and high-dose loganin groups were given the corresponding drugs by gavage once a day for eight weeks.At the end of the experiment,serum and liver samples were collected from each group of mice,and their body weights and liver weights were recorded,and liver indexes were calculated;serum viability of aspartate aminotransferase(AST),alanine aminotransferase(ALT)and triglyceride(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C),and low-density lipoprotein cholesterol(LDL-C)were detected,respectively.Meanwhile,the levels of TG,malondialdehyde(MDA),total-superoxide dismutase(T-SOD),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-1β(IL-1β)were detected in the livers of the mice in each group.Histopathological changes in the liver observed by hematoxylin-eosin(HE)staining and oil red O(ORO)staining.Transcriptome sequencing was used to screen differentially expressed genes in the liver of fructose-fed mice after loganin intervention,and functional enrichment analysis of differentially expressed genes was performed.Interaction of loganin between adenosine monophosphate activated protein kinaseα(AMPKα)protein was analysis by molecular docking.The expression levels of AMPKα,phosphorylated-AMPKα(p-AMPKα),sterol regulatory element binding protein-1(SREBP-1),and fatty acid synthetase(FASN)were detected by Western blotting,and the expression level of p-AMPKαprotein in the liver tissues of mice was detected by immunohistochemistry.Results Compared with the model group,the body weight,liver weight and liv

关 键 词：马钱苷 肝脏脂代谢紊乱 果糖饮食 AMPK信号通路 脂肪酸合成 

分 类 号：R285.5[医药卫生—中药学]