中草药2026，Vol.57Issue(4)：1377-1390,14.DOI:10.7501/j.issn.0253-2670.2026.04.016
基于UHPLC-MS技术及体内外实验探讨参附注射液对慢性心力衰竭心阳虚证大鼠微管蛋白翻译后修饰表达的影响
Effect of Shenfu Injection on tubulin post-translational modifications in rats with chronic heart failure of heart-yang deficiency syndrome based on UHPLC-MS and in vivo,in vitro experiments
摘要
Abstract
Objective To screen the core active components of Shenfu Injection(参附注射液,SFI)using UHPLC-MS and molecular docking techniques,and to verify the effects of SFI on expressions of post-translational modifications of tubulin in chronic heart failure of heart-yang deficiency syndrome through in vivo and in vitro experiments.Methods UHPLC-MS was applied to identify the main components of SFI,and molecular docking was performed to analyze their interactions with tubulin-related targets.SD rats were randomly divided into control group,model group,colchicine(0.1 mg/kg)group,SFI low-and high-dose(3,6 mL/kg)groups,with eight rats in each group.The CHF heart-yang deficiency model was replicated using sc isoproterenol,and drug intervention was given for two weeks.Echocardiography was used to detect cardiac function.ELISA was used to detect N-terminal pro-B-type natriuretic peptide(NT-proBNP)level in serum.Hematoxylin-eosin(HE)staining was used to detect pathological changes in myocardial tissue.Immunofluorescence was used to detect the expressions of acetyl α-tubulin,polyglut α-tubulin and detyr α-tubulin in myocardial tissue.Immunohistochemistry was used to detect the expressions of tubulin tyrosine ligase(TTL)and histone deacetylase 6(HDAC6)in myocardial tissue.H9c2 cardiomyocytes were treated with isoproterenol to establish a cell injury model,and after intervention with SFI,the level of microtubule protein modification was detected.Results A total of 12 main active ingredients in SFI were obtained through UHPLC-MS screening,and molecular docking results showed strong binding affinity with tubulin and modification related proteins.The animal experiment results showed that compared with model group,SFI could significantly improve cardiac function of rats(P<0.01),reduce the level of NT-proBNP in serum(P<0.01),alleviate the degree of myocardial interstitial fibrosis,reduce the expressions of microtubule protein α-tubulin and microtubule density in myocardial tissue(P<0.05,0.01),reduce the expressions of microtubule protein tyrosine and glutamate,increase acetylation expression,down-regulate HDAC6 positive expression(P<0.01),and up-regulate TTL positive expression(P<0.01).The results of cell experiments were consistent with animal experiments,further verifying that SFI could improve post-translational modifications of microtubule proteins.Conclusion SFI could effectively reduce the levels of tyrosinization and glutamatzation of microtubules,increase acetylation levels,correct the imbalance of post-translational modifications of microtubules,thereby improving the abnormal microtubule network in cardiac myocytes of rats with CHF heart-yang deficiency,enhancing microtubule stability and improving heart function.This provides experimental evidence for the molecular mechanism of SFI treatment for CHF heart-yang deficiency.关键词
参附注射液/慢性心力衰竭/微管蛋白/去酪氨酸化/谷氨酸化/乙酰化Key words
Shenfu Injection/chronic heart failure/tubulin/detyrosination/polyglutamylation/acetylation分类
医药卫生引用本文复制引用
李琳,叶嘉豪,赵震宇,李伟军,胡志希..基于UHPLC-MS技术及体内外实验探讨参附注射液对慢性心力衰竭心阳虚证大鼠微管蛋白翻译后修饰表达的影响[J].中草药,2026,57(4):1377-1390,14.基金项目
国家自然科学基金资助项目(82305092) （82305092）
国家自然科学基金资助项目(82574922) （82574922）
湖南中医药大学杏林英才支持计划(2025) （2025）
湖南省科技创新计划资助(2024RC3199) （2024RC3199）
湖南中医药大学优秀青年项目(Z2023XJYQ03) （Z2023XJYQ03）
