摘要
目的探讨咖啡酸(CA)对阿霉素(ADM)诱导的心肌细胞损伤的保护作用及其机制。方法采用不同浓度ADM诱导建立心肌细胞H9c2损伤模型,确定ADM诱导的最佳浓度。其次,在心肌损伤模型的基础上给予不同浓度CA,培养24 h确定最适CA浓度。将H9c2心肌细胞分为模型组(ADM组)、治疗组(ADM+CA组)和空白对照组(Control组),采用MTT法测定心肌细胞的存活率,用HE染色观察心肌细胞组织形态学变化。测定H9c2心肌细胞中乳酸脱氢酶(LDH)和活性氧(ROS)水平、线粒体膜电位变化及Na^(+)-K^(+)-ATP酶和Ca^(2+)-ATP酶活性,并运用Western blot检测NF-κB-p65的表达水平。结果选择2.0μmol/L ADM作为诱导浓度,并选择4.0μmol/L CA用于后续试验。与Control组比较,ADM组中心肌细胞纤维排列紊乱,肌纤维变形严重;LDH和ROS水平升高,线粒体膜电位升高,且Na^(+)-K^(+)-ATP酶和Ca^(2+)-ATP酶活性降低,NF-κB-p65蛋白表达增高。与ADM组比较,ADM+CA组中心肌细胞肌束排列整齐,LDH和ROS水平降低,线粒体膜电位降低(P<0.05),且线粒体Na^(+)-K^(+)-ATP和Ca^(2+)-ATP酶活性提高(P<0.05),NF-κB-p65蛋白表达降低(P<0.05)。结论CA通过抗氧化应激、降低线粒体膜电位、提高H9c2细胞内ROS水平,并且通过调控NF-κB信号通路,参与保护ADM诱导的心肌损伤过程。
Objective This study was to explore the protective effect and mechanism of caffeic acid on adriamycin-induced cardiomyocyte damage.Methods Adriamycin(ADM)is used to induce H9c2 injury model of cardiomyocytes and to determine the best concentration of it.Then the injured H9c2 were co-cultured with different concentrations of caffeic acid(CA)for 24 hours,and the optimal concentration of caffeic acid was selected.The H9c2 were divided into model group(ADM),treatment group(ADM+CA)and blank control group(control).As shown,the survival rate of cardiomyocytes was measured by MTT method,and the morphology of H9c2 was observed by HE.Then the levels of lactate dehydrogenase(LDH)and reactive oxygen species(ROS),changes in mitochondrial membrane potential and the activities of Na^(+)-K^(+)-ATPase and Ca^(2+)-ATPase in H9c2 were measured,as well as the expression of NF-κB-p65 were detected by Western blot.Results 2.0μmol/L ADM and 4.0μmol/L caffeic acid were selected for subsequent experiments.Compared with the control group,in the adriamycin-treated model group,the cardiomyocyte fiber arrangement was disordered,and the muscle fiber was deformed seriously;LDH and ROS levels were increased,mitochondrial membrane potential were increased,and Na^(+)-K^(+)-ATPase and Ca^(2+)-ATPase activities were reduced.Compared with the model group,in the caffeic acid treatment group,myocardial cell muscle bundles were arranged neatly,LDH and ROS levels,and mitochondrial membrane potential were reduced(P<0.05),while mitochondrial Na^(+)-K^(+)-ATP and Ca^(2+)-ATPase activities were increased(P<0.05),NF-κB-p65 protein expression was reduced(P<0.05).Conclusion Caffeic acid can protect against oxidative stress,reduce mitochondrial membrane potential,increase the level of reactive oxygen species in H9c2 cells,and participate in the protection of adriamycin-induced myocardial injury by regulating the NF-κB signaling pathway.
作者
唐炜
叶鹏林
刘坤
朱杨杰
高恶斌
Tang Wei;Ye Penglin;Liu Kun;Zhu Yangjie;Gao Ebin(Dept of Pediatrics,Affiliated Hospital of Jiangsu University,Zhengjiang 212000;School of Life Sciences, Jiangsu University,Zhengjiang 212000)
出处
《安徽医科大学学报》
CAS
北大核心
2022年第3期402-407,共6页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:31200019)
镇江市社会发展项目(编号:SH2019055)
江苏省妇幼保健项目(编号:F201648)
中国科学院南海海洋研究所开放基金(编号:LMB131001)。
关键词
咖啡酸
心肌毒性
氧化应激
线粒体
NF-ΚB
caffeic acid
myocardial toxicity
oxidation stress
mitochondria
NF-κB
