补肾降脂方调控TFEB-自噬-溶酶体途径干预ApoE^(-/-)小鼠动脉粥样硬化的机制  

Mechanism on Bushen Jiangzhi Formula in regulating TFEB-autophagy-lysosome pathway on atherosclerosis in ApoE^(-/-) mice

作　　者：贺平一 李琳丹 张悦[1,2] 申定珠 HE Pingyi;LI Lindan;ZHANG Yue;SHEN Dingzhu(Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Shanghai Geriatrics Institute of Chinese Medicine,Shanghai 200031,China)

机构地区：[1]上海中医药大学,上海201203 [2]上海市中医老年医学研究所,上海200031

出　　处：《中华中医药杂志》2025年第2期630-635,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(No.82074506,No.81873348);全国中医药创新骨干人才培训项目(No.国中医药人教函[2019]128号)。

摘　　要：目的:基于转录因子EB(TFEB)-自噬-溶酶体途径探讨补肾降脂方对ApoE^(-/-)动脉粥样硬化(AS)小鼠的效应机制。方法:100只雄性ApoE^(-/-)小鼠随机分为普食组、模型组、补肾降脂方组、阿托伐他汀组、3-甲基腺嘌呤(3-MA)组,每组20只;20只雄性C57BL/6J小鼠为正常组。给予相应药物干预12周后,HE、油红O染色分别观测小鼠主动脉窦病理形态与脂质蓄积情况;生化检测小鼠外周血血清总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白(HDL)、低密度脂蛋白(LDL)含量;免疫组化、免疫荧光分别观测小鼠主动脉窦TFEB蛋白表达与核转位;透射电子显微镜观察小鼠主动脉窦自噬小体与自噬溶酶体;Western Blot检测小鼠主动脉溶酶体相关膜蛋白1(LAMP1)、P62及微管相关蛋白轻链3(LC3)蛋白表达。结果:与模型组比较,补肾降脂方组主动脉窦可见自噬小体与自噬溶酶体,AS斑块与红染脂质面积显著减小(P<0.01),TC、TG、LDL含量显著下降(P<0.01),HDL含量显著升高(P<0.01),TFEB蛋白表达与核转位显著上调(P<0.01),LAMP1、LC3蛋白表达显著升高(P<0.01),P62蛋白表达显著降低(P<0.01)。结论:补肾降脂方可有效干预治疗AS,其机制与TFEB-自噬-溶酶体途径有关。Objective:To explore the effect and mechanism of Bushen Jiangzhi Formula on ApoE^(-/-) mice of atherosclerosis(AS)based on transcription factor EB(TFEB)-autophagy-lysosome pathway.Methods:A total of 100 male ApoE^(-/-)mice were randomly divided into 5 groups:common diet group,model group,Bushen Jiangzhi Formula group,atorvastatin group(ATV)and autophagy inhibitor group(3-MA),20 in each,with 20 male C57BL/6J mice as normal group.After 12 weeks of corresponding drug intervention,the pathological morphology and lipid accumulation of aortic sinus in mice were observed by HE and oil red O staining;contents of total cholesterol(TC),triglyceride(TG),high-density lipoprotein(HDL)and low-density lipoprotein(LDL)in peripheral blood of mice were detected by biochemical assay;the protein expression and nuclear translocation of TFEB in aortic sinus were observed by immunohistochemistry and immunofluorescence;the autophagosomes and autophagy lysosomes were observed by transmission electron microscope;the proteins expressions of lysosome-associated membrane proteins(LAMP1),P62 and microtubule-associated proteins light chain 3(LC3)in mouse aorta were detected by Western Blot.Results:Compared with model group,autophagosomes and autophagy lysosomes were found in Bushen Jiangzhi Formula group;the area of AS plaque and red-stained lipid in aortic sinus decreased significantly(P<0.01);the contents of TC,TG,LDL decreased significantly(P<0.01)while HDL increased significantly(P<0.01);the protein expression and nuclear translocation of TFEB upregulated significantly(P<0.01);the protein expression of LAMP1 and LC3 increased significantly(P<0.01)while the P62 decreased significantly(P<0.01).Conclusion:Bushen Jiangzhi Formula can effectively interfere with the treatment of AS,whose mechanism is related to regulating TFEB-autophagy-lysosome pathway.

关 键 词：补肾降脂方 动脉粥样硬化 转录因子EB 自噬 溶酶体 

分 类 号：R28[医药卫生—中药学]