文献类型：期刊文献

中文题名：麝香通心滴丸对心肌细胞缺氧/复氧损伤的保护作用

英文题名：Protective effect of shexiang tongxin dropping pills on anoxia-reoxygenation injury in H9c2 cardiomyocytes

作者：刘新文[1];沈男男[1];王佳良[1];傅永平[2]

机构：[1]绍兴文理学院附属医院(绍兴市立医院)药剂科,浙江绍兴312000;[2]绍兴文理学院附属医院(绍兴市立医院)心血管内科,浙江绍兴312000

年份：2020

卷号：13

期号：3

起止页码：170

中文期刊名：中华危重症医学杂志：电子版

收录：CSTPCD、、CSCD_E2019_2020、CSCD

基金：浙江省自然科学基金项目(LY18H020008);浙江省医药卫生科技计划项目(2018KY842、2019RC297);绍兴市科技计划项目(2018C30031)。

语种：中文

中文关键词：肌细胞;心脏;缺氧;麝香通心滴丸

外文关键词：Myocytes,cardiac;Anoxia;Shexiang tongxin dropping pill

中文摘要：目的探讨麝香通心滴丸(STDP)对心肌细胞缺氧/复氧的保护作用。方法建立心肌缺氧/复氧模型。将H9c2细胞分为5组:对照组(细胞正常培养,不做任何处理)、模型组、STDP组(在复氧前40 min给予STDP 50 mg/L)、3-甲基腺嘌呤(3-MA)组(在复氧前40 min给予3-MA 5 mmol/L)及STDP+3-MA组(在复氧前40 min分别给予STDP 50 mg/L及3-MA 5 mmol/L)。采用细胞计数试剂盒8(CCK-8)法检测各组细胞存活率,并测定各组细胞乳酸脱氢酶(LDH)、丙二醛及肌酸激酶水平。采用流式细胞仪检测各组细胞的细胞凋亡率,实时荧光定量PCR检测信使RNA(mRNA)自噬相关基因Beclin-1、Atg5的表达水平,并通过Western-blotting检测各组细胞自噬相关蛋白LC3Ⅱ/LC3Ⅰ、低氧诱导因子1α(HIF-1α)、Bcl-2/腺病毒E1B 19 kDa相互作用蛋白3(BNIP3)、Beclin-1、Atg5的表达水平。结果各组细胞存活率比较,差异有统计学意义(F=85.893,P=0.028),且STDP组细胞存活率较模型组、3-MA组及STDP+3-MA组细胞存活率均显著升高[(90±7)%、(63±5)%、(80±5)%、(81±6)%,P均<0.05]。各组细胞LDH、丙二醛、肌酸激酶水平,细胞凋亡率,Beclin-1 mRNA、Atg5 mRNA以及LC3Ⅱ/LC3Ⅰ、HIF-1α、BNIP3、Beclin-1、Atg5蛋白表达水平的比较,差异均有统计学意义(F=78.381、23.519、48.376、22.726、6.315、5.294、75.219、116.546、21.125、39.724、65.247,P均<0.05)。进一步两两比较发现,在STDP组细胞中,LDH[(92±6)、(194±13)、(195±11)、(192±10)μmol/L,P均<0.05]、丙二醛[(12.5±0.7)、(17.2±1.2)、(18.5±1.6)、(17.9±1.0)μmol/L,P均<0.05]、肌酸激酶[(19.9±1.0)、(34.3±2.2)、(35.3±2.2)、(34.8±2.5)μmol/L,P均<0.05]水平,细胞凋亡率[(5.8±0.8)%、(8.8±0.9)%、(7.6±0.7)%、(7.3±0.5)%,P均<0.05]较模型组、3-MA组及STDP+3-MA组均明显降低;Beclin-1 mRNA、Atg5mRNA、LC3Ⅱ/LC3Ⅰ、BNIP3、Beclin-1、Atg5蛋白表达水平较模型组均明显降低,而较3-MA组及STDP+3-MA组均显著升高(P均<0.05);HIF-1α蛋白表达水平较模型组、3-MA组及STDP+3-MA组均明显升高(P均<0.05)。结论STDP可通过调控HIF-1α/BNIP3信号通路发挥对心肌细胞缺氧/复氧损伤的保护作用。

外文摘要：Objective To study the protective effect of shexiang tongxin dropping pills(STDP)on anoxia-reoxygenation injury in H9c2 cardiomyocytes.Methods A model of myocardial anoxia-reoxygenation was established and the H9c2 cardiomyocytes were divided into 5 groups:a control group(with normal H9c2 cardiomyocytes),a model group,a STDP group(given 50 mg/L STDP 40 min before reoxygenation),a 3-methyladenine(3-MA)group(given 5 mmol/L 3-MA 40 min before reoxygenation),and a STDP+3-MA group(given 50 mg/L STDP+5 mmol/L 3-MA 40 min before reoxygenation).The cell survival rate was detected by cell counting kit-8,and the levels of lactate dehydrogenase(LDH),malonaldehyde,and creatine kinase were compared among the five groups.The apoptosis rate of cardiomyocytes was detected by flow cytometry.The messenger RNA(mRNA)expression levels of Beclin-1 and Atg5 were examined by real-time fluorescent quantitative PCR.The protein expression levels of LC3Ⅱ/LC3Ⅰ,hypoxia-inducible factor-1alpha(HIF-1α),Bcl-2/adenovirus E1B 19 kDa-interacting protein 3(BNIP3),Beclin-1,and Atg5 were detected by Western-blotting.Results The cell survival rate was significantly different among the five groups(F=85.893,P=0.028),and it was much higher in the STDP group than in the model group,3-MA group and STDP+3-MA group[(90±7)%,(63±5)%,(80±5)%,(81±6)%;all P<0.05].The LDH,malonaldehyde,creatine kinase,apoptosis rate,Beclin-1 mRNA,Atg5 mRNA,LC3Ⅱ/LC3Ⅰprotein,HIF-1αprotein,BNIP3 protein,and Beclin-1 protein were significantly different among the five groups(F=78.381,23.519,48.376,22.726,6.315,5.294,75.219,116.546,21.125,39.724,65.247;all P<0.05).In the STDP group,the levels of LDH[(92±6),(194±13),(195±11),(192±10)μmol/L],malonaldehyde[(12.5±0.7),(17.2±1.2),(18.5±1.6),(17.9±1.0)μmol/L],and creatine kinase[(19.9±1.0),(34.3±2.2),(35.3±2.2),(34.8±2.5)μmol/L]and the apoptosis rate of cardiomyocytes[(5.8±0.8)%,(8.8±0.9)%,(7.6±0.7)%,(7.3±0.5)%]were much lower than those in the model group,3-MA group and STDP+3-MA group,and the HIF-1αprotein level was much higher(all P<0.05);meanwhile,the levels of Beclin-1 mRNA,Atg5 mRNA,LC3Ⅱ/LC3Ⅰprotein,BNIP3 protein,Beclin-1 protein,and Atg5 protein were much lower than those in the model group,but were much higher than those in the 3-MA group and STDP+3-MA group(all P<0.05).Conclusion The STDP can protect cardiomyocytes from anoxia-reoxygenation injury by regulating the HIF-1α/BNIP3 signaling pathway.