摘要
目的探讨芍药内酯苷改善缺氧/复氧(hypoxia/reoxygenation,H/R)诱导的心肌细胞损伤的作用机制。方法H/R诱导大鼠H9c2心肌细胞建立心肌缺血再灌注损伤模型,给予芍药内酯苷干预,检测丙二醛(malondialdehyde,MDA)水平及超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性;流式细胞术测定细胞凋亡率;Western blotting检测凋亡相关蛋白表达;qRT-PCR检测miR-219a-5p表达。转染miR-219a-5p mimics或anti-miR-219a-5p,考察下调miR-219a-5p对芍药内酯苷改善H/R诱导的心肌细胞损伤作用的影响。体内建立缺血再灌注(ischemia/reperfusion,I/R)小鼠模型,给予芍药内酯苷干预,检测血清中肌酸激酶同工酶(creatine kinase isoenzyme,CKMB)活性和心肌肌钙蛋白T(cardiac troponin T,c TnT)水平;检测心肌组织MDA水平及SOD、GSH-Px活性;采用苏木素-伊红(HE)染色考察心肌组织病理变化;采用TTC染色检测心肌组织梗死情况。结果与对照组比较,模型组H9c2细胞MDA水平、细胞凋亡率及剪切型半胱氨酸天冬氨酸蛋白酶-3(cleaved cystein-asparate protease-3,cleaved Caspase-3)、cleaved Caspase-9蛋白表达水平显著上升(P<0.05),而miR-219a-5p表达、SOD、GSH-Px活性显著下降(P<0.05);给予芍药内酯苷干预后,MDA水平、细胞凋亡率及凋亡相关蛋白表达显著降低(P<0.05),miR-219a-5p表达、SOD、GSH-Px活性显著升高(P<0.05),且呈剂量相关性;下调miR-219a-5p表达后,显著逆转了芍药内酯苷对H/R诱导的心肌细胞损伤的抑制作用(P<0.05)。体内结果与体外结果一致,芍药内酯苷显著缓解了I/R引发的小鼠心肌组织损伤和凋亡。结论芍药内酯苷通过调控miR-219a-5p表达而抑制心肌细胞氧化应激及凋亡,从而减轻H/R诱导的心肌细胞损伤。
Objective To explore the mechanism of albiflorin on improving hypoxia/reoxygenation(H/R)-induced myocardial cell injury.Methods H/R was used to induce H9c2 cardiomyocytes to establish a myocardial ischemia-reperfusion injury model.Albiflorin was used to intervene,the level of malondialdehyde(MDA)and activities of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)were detected;Flow cytometry was used to measure the apoptosis rate of cells;Western blotting was used to detect the expressions of apoptosis-related proteins;qRT-PCR was used to detect the expression of miR-219a-5p.miR-219A-5p mimetics or anti-miR-219a-5p was transfected to investigate the effect of down-regulating of miR-219a-5p on improvement of H/Rinduced myocardial cell injury by albiflorin.An ischemia-reperfusion(I/R)mouse model in vivo was established,albiflorin was used to intervene,creatine kinase isoenzyme(CK-MB)activity and cardiac troponin T(cTnT)level in serum were detected;Levels of MDA and activities of SOD and GSH-Px in myocardial tissue were detected;Hematoxylin eosin(HE)staining was used to investigate pathological changes in myocardial tissue;TTC staining was used to detect myocardial tissue infarction.Results Compared with control group,MDA level,apoptosis rate,and expression levels of cleaved cysteine aspartate protease-3(cleaved Caspase-3)and cleaved Caspase-9 proteins in H9c2 cells of model group were significantly increased(P<0.05),while the expression of miR-219a-5p,SOD and GSH Px activities were significantly decreased(P<0.05);After intervention with albiflorin,MDA level,apoptosis rate,and expressions of apoptosis related proteins were significantly reduced(P<0.05),while miR-219a-5p expression,SOD and GSH Px activity were significantly increased(P<0.05),and showed a dose-dependent relationship;After down-regulating the expression of miR-219a-5p,the inhibitory effect of albiflorin on H/R-induced myocardial cell injury was significantly reversed(P<0.05).The in vivo and in vitro results were consistent,albiflorin significantly alleviated myocardial tissue damage and apoptosis induced by I/R.Conclusion Albiflorin inhibits oxidative stress and apoptosis of myocardial cells by regulating the expression of miR-219a-5p,thereby alleviating H/R-induced myocardial cell damage.
作者
李霞
王成
侯永兰
王建美
金卫东
LI Xia;WANG Cheng;HOU Yonglan;WANG Jianmei;JIN Weidong(Department of Cardiovascular Medicine,Xinxiang Central Hospital,Xinxiang 453000,China)
出处
《中草药》
北大核心
2025年第12期4327-4335,共9页
Chinese Traditional and Herbal Drugs
基金
河南省医学科技攻关计划项目(LHGJ20230886)。
