| 于梦,宋欣丽,孙丽,等.大血藤总酚酸调控微小 RNA-155对缺氧复氧心肌细胞的细胞活性凋亡及氧化应激的影响[J].安徽医药,2021,25(12):2355-2359. |
| 大血藤总酚酸调控微小 RNA-155对缺氧复氧心肌细胞的细胞活性凋亡及氧化应激的影响 |
| Effect of Sargentodoxa cuneatatotal phenolic acid on cell activity, apoptosis and oxidative stress of hypoxia-reoxygenation cardiomyocytes by regulating miR-155 |
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| DOI：10.3969/j.issn.1009-6469.2021.12.006 |
| 中文关键词: 心肌再灌注损伤 大血藤总酚酸 miR-155 肌酸激酶 L-乳酸脱氢酶 丙二醛 缺氧复氧 心肌细胞 凋亡 氧化应激 |
| 英文关键词: Myocardial reperfusion injury Sargentodoxa cuneatatotal phenolic acid MiR-155 Creatine kinase L-lactate de. hydrogenase Malondialdehyde Hypoxia and reoxygenation Cardiomyocytes Apoptosis Oxidative stress |
| 基金项目:北京市自然科学基金( 7172124) |
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| 中文摘要: |
| 目的探讨大血藤总酚酸对缺氧复氧心肌细胞的细胞活性凋亡及氧化应激的影响及分子机制。方法 2019年 12月至 2020年 8月，心肌细胞 H9C2分为对照组(正常培养)、缺氧复氧组(缺氧 4h，复氧 12 h)、大血藤总酚酸低、中、高浓度组(造模前分别用 12.5、25.0、50.0 mg/L大血藤总酚酸培养 2h，然后进行缺氧复氧处理)、大血藤总酚酸 -H+NC组(将 miR-155模拟物阴性对照转染至 H9C2中再用 50.0 mg/L大血藤总酚酸培养，最后进行缺氧复氧处理)、大血藤总酚酸 -H+miR-155 mimic组(将 miR-155模拟物转染至 H9C2中再用 50.0 mg/L大血藤总酚酸培养，最后进行缺氧复氧处理)。实时荧光定量 PCR(RT-qPCR)检测微小 RNA-155(miR-155)表达水平； MTT法检测细胞存活率；流式细胞术检测细胞凋亡；蛋白质印迹法(Western blotting)检测蛋白表达；试剂盒检测细胞培养液中肌酸激酶、乳酸脱氢酶( LDH)、丙二醛含量。结果缺氧复氧处理的心肌细胞中 miR-155表达水平升高，细胞吸光度降低，心肌细胞凋亡率升高，活化胱天蛋白酶 -3(cleaved-caspase-3)表达水平升高，胱天蛋白酶 -3前体(pro-caspase-3)表达水平降低，心肌细胞中肌酸激酶、 LDH、丙二醛含量升高(P<0.05)。大血藤总酚酸中、高浓度处理后缺氧复氧诱导的心肌细胞中 miR-155表达水平降低，细胞吸光度升高，心肌细胞凋亡率降低， cleaved-caspase-3表达水平降低， pro-caspase-3表达水平升高，心肌细胞中肌酸激酶、 LDH、丙二醛含量降低，且呈浓度依赖性( P<0.05)；而大血藤总酚酸低浓度组各数据无显著变化。过表达 miR-155可逆转大血藤总酚酸对缺氧复氧诱导的心肌细胞活性、凋亡［( 22.73±0.58)%比( 13.55±0.33)%，P<0.05］和肌酸激酶［( 79.19±2.78)U/L比( 41.22±2.31)U/L，P<0.05］、 LDH［( 44.20±2.80)U/L比( 24.77±2.46)U/L，P<0.05］、丙二醛［( 24.05±1.28)μmol/L比( 12.73±0.59)μmol/L，P<0.05］含量的影响。结论大血藤总酚酸可能通过下调 miR-155抑制缺氧复氧心肌细胞的凋亡及氧化应激。 |
| 英文摘要: |
| Objective To investigate the effect of Sargentodoxa cuneatatotal phenolic acid on cell activity, apoptosis and oxidative stress of hypoxia-reoxygenation cardiomyocytes and its molecular mechanism.Methods From December 2019 to August 2020, cardio. myocytes H9C2 were assigned into control group (normal culture), hypoxia-reoxygenation group (hypoxia 4 h, reoxygenation 12 h), sar.gentodoxa cuneatatotal phenolic acid low, medium and high concentration group (12.5, 25.0, 50.0 mg/L of Sargentodoxa cuneatatotalphenolic acid were cultured for 2 h before modeling, and then hypoxia and reoxygenation treatment), sargentodoxa cuneatatotal pheno.lic acid-H+NC group (miR-155 mimic negative control was transfected into H9C2 and then cultured with 50.0 mg/L of Sargentodoxa cu.neatatotal phenolic acid, and finally treated with hypoxia and reoxygenation), sargentodoxa cuneatatotal phenolic acid-H+miR-155 mim. ic group (miR-155 mimic was transfected into H9C2 and then cultured with 50.0 mg/L of Sargentodoxa cuneatatotal phenolic acid, andfinally treated with hypoxia and reoxygenation). Real-time fluorescence quantitative PCR (RT-qPCR) was used to detect the expression of miR-155; the tetramethylazolium salt colorimetric method (MTT) was used to detect cell viability; flow cytometry was used to detectcell apoptosis; Western blotting method was used to detect protein expression; kit was used to detect the content of CK, LDH, MDA in cell culture fluid.Results The expression of miR-155 in the cardiomyocytes treated with hypoxia and reoxygenation was increased,the cell OD value was decreased, the apoptosis rate of cardiomyocytes was increased, the expression of cleaved-caspase-3 was in. creased, the expression of pro-caspase-3 was decreased, and the contents of CK, LDH and MDA were increased (P<0.05). The expres. sion of miR-155 in cardiomyocytes induced by hypoxia and reoxygenation was decreased after treatment with middle and high concen.tration of sargentodoxa cuneatatotal phenolic acid, the cell OD value was increased, the apoptosis rate of cardiomyocytes was decreased,the expression of cleaved-caspase-3 was decreased, and the expression of pro-caspase-3 was increased, and the content of CK, LDH, and MDA in cardiomyocytes were decreased, in a concentration-dependent manner (P<0.05); however, the data of the low-concentra. tion group of Sargentodoxa cuneatatotal phenolic acid did not change significantly. Overexpression of miR-155 reversed the effect of Sargentodoxa cuneatatotal phenolic acid on cardiomyocyte activity, apoptosis [(22.73±0.58)% vs. (13.55±0.33)% , P<0.05] and CK [(79.19±2.78)U/L vs. (41.22±2.31)U/L, P<0.05], LDH [(44.20±2.80)U/L vs. (24.77±2.46)U/L, P<0.05] and MDA [(24.05±1.28)μmol/L vs. (12.73±0.59)μmol/L, P<0.05] content induced by hypoxia and reoxygenation.Conclusion Sargentodoxa cuneatatotal phenolic acid may inhibit hypoxia-reoxygenation cardiomyocyte apoptosis and oxidative stress by down-regulating miR-155. |
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