北葶苈子炮制前后对H2O2诱导的H9c2心肌细胞损伤保护作用  

Protective effects of differently treated Lepidium apetalum seeds on HO-induced H9c2 cardiomyocytes injury

作　　者：王慧慧[1] 张莉[1] 杨方方[1] 冯卫生[1] 郑晓珂[1] WANG Hui-hui;ZHANG Li;YANG Fang-fang;FENG Wei-sheng;ZHENG Xiao-ke(Henan University of Chinese Medicine,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学,河南郑州450046

出　　处：《中成药》2020年第8期2018-2024,共7页Chinese Traditional Patent Medicine

基　　金：中央引导地方科技发展专项(14104349);河南省重大科技专项(171100310500);河南中医药大学2018年度博士科研基金(BSJJ2018-04)。

摘　　要：目的比较北葶苈子炮制前后对H2O2诱导的H9c2心肌细胞损伤的保护作用。方法H2O2诱导H9c2细胞建立细胞损伤模型,同时加入北葶苈子生品及炮制品(400、200、100μg/mL)处理细胞6 h。MTT法检测H9c2细胞存活率,流式细胞术检测细胞ROS、线粒体膜电位、细胞凋亡率、自噬空泡,Incell-Western法检测细胞凋亡相关蛋白,并检测细胞LDH、MDA、T-SOD、GSH-PX水平。结果与模型组比较,北葶苈子炮制前后均能降低H9c2细胞凋亡率、自噬空泡、ROS水平及凋亡相关蛋白Bax/Bcl-2比值(P<0.01),增加线粒体膜电位(P<0.01),降低细胞培养液中LDH及细胞MDA水平(P<0.01),提高细胞T-SOD、GSH-PX水平(P<0.05,P<0.01);北葶苈子炮制品抑制caspase-3蛋白表达(P<0.05)。结论北葶苈子炮制前后均可有效改善由H2O2诱导的心肌细胞损伤,可能与其调节氧化应激的作用有关。AIM To compare the protective effects of crude and processed Lepidium apetalum Willd.(L.apetalum)seeds(LaSs)on H2O2-induced H9 c2 cardiomyocytes(HHCs)injury.METHODS H9 c2 cells injury models established by H2O2 induction were exposed to 6 h crude and processed LaSs(400,200,100μg/mL)differently.H9 c2 cells were subjected to survival rate identification by MTT method,detection of cellular ROS,mitochondrial membrane potential,apoptosis rate,autophagic vacuole by flow cytometry,detection of apoptosis-related proteins by Incell-Western method,and levels of LDH,MDA,T-SOD and GSH-PX.RESULTS Compared with the model group,both groups of crude and processed LaSs shared reduced H9 c2 cell apoptosis rate,autophagic vacuole,ROS level and apoptosis-related protein Bax/Bcl-2 ratio(P<0.01),increased mitochondrial membrane potential(P<0.01),reduced the LDH level in the cell culture medium and the cell MDA level(P<0.01),and increased levels of T-SOD and GSH-PX(P<0.05,P<0.01).In addition,processed LaSs inhibited the expression of caspase-3 protein(P<0.05).CONCLUSION Both the crude and processed LaSs can ameliorate the myocardial cell injury induced by H2O2 effectively,which may be related to the enhancement of oxidative stress inhibition.

关 键 词：北葶苈子 H9C2细胞 凋亡 自噬 氧化应激 

分 类 号：R285.5[医药卫生—中药学]