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.
作者
王慧慧
张莉
杨方方
冯卫生
郑晓珂
WANG Hui-hui;ZHANG Li;YANG Fang-fang;FENG Wei-sheng;ZHENG Xiao-ke(Henan University of Chinese Medicine,Zhengzhou 450046,China)
出处
《中成药》
CAS
CSCD
北大核心
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)。
关键词
北葶苈子
H9C2细胞
凋亡
自噬
氧化应激
Lepidium apetalum Willd.Seeds(LaSs)
H9c2 cells
apoptosis
autophagy
oxidative stress
