北葶苈子中抗心肌细胞凋亡活性成分筛选  

Screening of anti-myocardial cell apoptosis active components in the seeds of Lepidium apetalum Willd.

作　　者：李静阳 张莉[1] 杨方方[1] 卢仁睿 李孟[1] 冯卫生[1,2] 郑晓珂 LI Jing-yang;ZHANG Li;YANG Fang-fang;LU Ren-rui;LI Meng;FENG Wei-sheng;ZHENG Xiao-ke(School of Pharmacy,Henan University of Chinese Medicine,Zhengzhou 450046,China;Henan Engineering Technology Research Center for Chinese Medicine Development,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学药学院,郑州450046 [2]河南省中药开发工程技术研究中心,郑州450046

出　　处：《中国新药杂志》2024年第18期1933-1942,共10页Chinese Journal of New Drugs

基　　金：国家重点研发计划-中医药现代化研究项目:基于病势在表在里的麻黄和葶苈子升降浮沉药性研究(2019YFC1708802);河南省科技攻关项目(212102310345);河南中医药大学2018年度博士科研基金资助项目(BSJJ2018-04)。

摘　　要：目的:从北葶苈子中提取、分离得到的27种化合物中筛选出对阿霉素诱导的心肌细胞损伤模型有改善作用的成分,并对其作用机制进行初步研究。方法:阿霉素梯度浓度给药筛选最佳给药浓度,采用5μg·mL-1阿霉素建立H9c2心肌细胞损伤模型,用北葶苈子中化合物处理细胞24 h,检测细胞活力及乳酸脱氢酶(lactate dehydrogenase,LDH)、总超氧化物歧化酶(superoxide dismutase,T-SOD)、丙二醛(malondialdehyde,MDA)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)水平,流式细胞术检测活性氧(reactive oxygen species,ROS)、线粒体膜电位和细胞凋亡水平,In-Cell-Western法检测线粒体凋亡通路关键蛋白表达水平。结果:北葶苈子中的北葶新苷D(LS-18)、槲皮素-3-O-β-D-葡萄糖-(1→2)-β-D-葡萄糖苷(LS-22)和槲皮素-3-O-β-D-吡喃半乳糖苷(LS-26)可有效提高心肌细胞活力,降低LDH,MDA水平,升高T-SOD水平,改善氧化应激。进步一研究发现LS-18,LS-22和LS-26可降低细胞ROS水平,提高线粒体膜电位水平,抑制心肌细胞凋亡,抑制线粒体凋亡通路关键蛋白半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)表达水平,降低B淋巴细胞瘤-2相关X蛋白/B淋巴细胞瘤-2(Bcl-2-associated X protein/B lymphocytoma-2,Bax/Bcl-2)水平。结论:北葶苈子中化合物LS-18,LS-22和LS-26对阿霉素诱导的H9c2细胞损伤具有显著的保护作用,其机制可能与改善氧化应激稳态、抑制线粒体凋亡途径有关。Objective:To screen the components with ameliorating effects on doxorubicin-induced cardiomyocyte injury model among the 27 compounds obtained by extraction and isolation from the seeds of Lepidium apetalum Willd.and conduct preliminary studies on their mechanisms of action.Methods:The optimal concentration of doxorubicin was screened,5μg·mL-1 was used to establish H9c2 cardiomyocyte injury model,and the cells were treated with compounds in the seeds of Lepidium apetalum Willd.for 24 h,then cell viability,LDH,T-SOD,MDA,GSH-Px levels were measured.ROS,mitochondrial membrane potential and apoptosis rate were detected by flow cytometry assay.In-Cell-Western assay was used to detect the expression levels of key proteins in the mitochondrial apoptotic pathway.Results:Lepidiumoside D(LS-18),quercetin3-O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranoside(LS-22),quercetin-3-O-β-D-galactopyranoside(LS-26)from the seeds of Lepidium apetalum Willd.could effectively enhance cell viability,reduce LDH,MDA levels,increase T-SOD,GSH-px levels,and improve oxidative stress.Further research showed LS-18,LS-22,LS-26 could decrease cell ROS level,enhance mitochondrial membrane potential level,inhibit apoptosis rate of cardiomyocytes and the expression level of the key protein Caspase-3 of the mitochondrial apoptosis pathway,and reduce the level of Bax/Bcl-2.Conclusion:Components LS-18,LS-22,LS-26 from the seeds of Lepidium apetalum Willd.have significant protective effect on H9c2 cardiomyocyte in doxorubicin-induced injury,and the mechanism may be related to improve oxidative stress homeostasis and inhibit the mitochondrial apoptotic pathway.

关 键 词：北葶苈子 北葶新苷 阿霉素 心肌损伤 线粒体凋亡通路 氧化应激 

分 类 号：R965[医药卫生—药理学]