芹菜素对化学法氧糖剥夺/复氧复糖损伤大鼠心肌细胞保护作用研究  被引量:2

The protective effect of apigenin on cardiomyocytes of rats injured by chemical oxygen-glucose deprivation/restoration

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作  者:禹博 祁琳 金鑫[3] 曹清文 田晨 李佳颖 王越 周建妹[5] 陈康寅[1] YU Bo;QI Lin;JIN Xin;CAO Qing-wen;TIAN Chen;LI Jia-ying;WANG Yue;ZHOU Jian-mei;CHEN Kang-yin(Department of Cardiology,The Second Hospital,Tianjin Medical University,Tianjin Key Laboratory of Ionic-Molecular Function of Cardiovascular Disease,Tianjin Institute of Cardiology,Tianjin 300211,China;Department of Pharmacy,Characteristic Medical Center of PAP,Tianjin 300162,China;Military Medicine Section,Logistics University of Chinese People′s Armed Police Force,Tianjin 300309,China;College of Integrated Traditional Chinese and Western Medicine,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;Department of Cadre Health Care,Zhejiang Hospital,Hangzhou 310013,China)

机构地区:[1]天津医科大学第二医院心脏科,天津市心血管病离子与分子机能重点实验室,天津心脏病学研究所,天津300211 [2]武警特色医学中心药剂科,天津300162 [3]武警后勤学院军事药学教研室,天津300309 [4]天津中医药大学中西医结合学院,天津301617 [5]浙江医院干部保健科,杭州310013

出  处:《天津医科大学学报》2023年第3期258-264,共7页Journal of Tianjin Medical University

基  金:浙江省医药卫生科技计划项目(2021KY013);天津市医学重点学科(专科)建设资助项目(TJYXZDXK-029A);天津中医药大学研究生创新基金(ZXYCXLX202119);天津中医药大学研究生创新基金(ZXYCXLX202018);武警部队创新研究项目(ZZKY20222419);武警后勤学院基础研究项目(WHJ202007)。

摘  要:目的:研究芹菜素(API)对化学法氧糖剥夺/复氧复糖(OGD/R)大鼠H9c2心肌细胞损伤的保护作用及分子机制。方法:首先给予H9c2细胞不同浓度的氯化钴(CoCl_(2))或1%O2处理24 h,通过测定细胞活力和低氧诱导因子-1α(HIF-1α)的表达水平,确定模拟1%O2的等效CoCl_(2)浓度;然后给予H9c2细胞CoCl_(2)或1%O_(2)处理合并无糖培养16 h,再正常培养(复氧复糖)4 h,测定细胞活力和HIF-1α的表达水平,确定OGD/R模型等效CoCl_(2)浓度。MTT法测定不同浓度API对OGD/R大鼠H9c2细胞活力的影响;ELISA法检测细胞Caspase-3含量;超氧化物歧化酶(SOD)和乳酸脱氢酶(LDH)试剂盒测定细胞SOD和LDH含量;采用Western印迹方法检测细胞Bcl-2、Bax、SIRT1、p62和LC3Ⅱ/Ⅰ表达水平。结果:筛选1%O_(2)的等效CoCl_(2)浓度结果显示,与对照组相比,CoCl_(2)(0.8 mmol/L)组细胞活力明显降低(P<0.05),HIF-1α表达明显上调(P<0.05)且与1%O_(2)组无显著差异(P>0.05),因此选取CoCl_(2)(0.8 mmol/L)用于OGD/R模型。与对照组相比,OGD/R组的细胞活力及SOD含量明显下降(P<0.05),LDH漏出量和Caspase-3含量明显提高(P<0.05),SIRT1、LC3Ⅱ/Ⅰ及Bcl-2表达量均明显下降(均P<0.05),Bax及p62的表达量明显增加(P<0.05);与OGD/R组相比,API(0.01~10μmol/L)组细胞活力有显著提高(P<0.05);10μmmol/L API能显著提高细胞SOD含量(P<0.05),显著降低LDH漏出量和Caspase-3含量(均P<0.05),明显提高SIRT1、LC3Ⅱ/Ⅰ及Bcl-2的表达量(均P<0.05),降低Bax及p62的表达量(均P<0.05)。结论:0.8 mmol/L CoCl_(2)合并剥糖16 h后恢复4 h可构建稳定的H9c2细胞OGD/R损伤模型,API可通过调节SIRT1增强自噬以抵抗H9c2细胞OGD/R损伤和凋亡。Objective:To investigate the protective effect andmolecularmechanismof apigenin(API)onH9c2 cardiomyocytes of rats injured by chemical glucose-oxygen deprivation/restoration(OGD/R).Methods:Firstly,H9c2 cells were treated with different concentrations of cobalt chloride(CoCl_(2))or 1%O2 for 24 h.The cell viability and the expression level of hypoxia-inducible factor-1α(HIF-1α)were measured to determine the equivalent CoCl_(2)concentration to simulate 1%O_(2).Then H9c2 cells were treated with CoCl_(2)or 1%O_(2) and cultured without glucose for 16 h,and then cultured normally(restoration)for 4 h.Cell viability and HIF-1αexpression level were measured to determine the equivalent CoCl_(2)concentration of the OGD/R model.MTT assay was used to determine the effects of different concentrations of API on the viability of H9c2 cells injured by OGD/R.The content of Caspase-3 was detected by ELISA.Superoxide dismutase(SOD)and lactate dehydrogenase(LDH)kits were used to determine the contents of SOD and LDH.The expression levels of Bcl-2,Bax,SIRT1,p62 and LC3Ⅱ/Ⅰwere detected by Wes tern blotting.Results:The results of screening the equivalent CoCl_(2)concentration to simulate 1%O2 showed that compared with the control group,the cell viability of CoCl_(2)(0.8 mmol/L)group was significantly decreased and the HIF-1αexpression was significantly up-regulated(P<0.05);and there was no significant difference between 1%O_(2) and the above results.Therefore,CoCl_(2)(0.8 mmol/L)was selected for OGD/R model.Compared with the control group,the cell viability and SOD content of the OGD/R group were significantly decreased(P<0.05);LDH leakage and Caspase-3 content in the OGD/R group were sig-nificantly increased(P<0.05);the expressions of SIRT1,LC3Ⅱ/Ⅰand Bcl-2 in OGD/R group were significantly decreased(all P<0.05),the expressions of Bax and p62 were significantly increased(both P<0.05).Compared with the OGD/R group,the API(0.01-10μmol/L)group significantly increased cell viability(P<0.05),10μmol/L API could significantly incre

关 键 词:芹菜素 氧糖剥夺/复氧复糖 氯化钴 心肌细胞 

分 类 号:R541[医药卫生—心血管疾病]

 

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