摘要： 目的 观察microRNA-486(miR-486)对原代心肌细胞低氧损伤过程的影响。方法 培养原代乳鼠心肌细胞,构建模拟心肌细胞急性缺血的低氧损伤模型,通过实时荧光定量PCR测定低氧培养6、12、24、36、48 h和常氧培养后miR-486的表达水平。另外,按照转染Ad-miR-486和Ad-miR-486-antago,分为常氧组(C+N组)、Ad-miR-486+常氧组(M+N组)、Ad-miR-486-antago+常氧组(I+N组)、低氧组(C+H组)、Ad-miR-486+低氧组(M+H组)、Ad-miR-486-antago+低氧组(I+H组)。通过CCK-8、乳酸脱氢酶(LDH)、流式细胞仪测定细胞活力、损伤和凋亡程度。结果 随着低氧损伤时间的延长,miR-486的表达逐渐上升,至24 h达到峰值后逐渐下降。常氧环境中,M+N组、I+N组与C+N组比较,细胞活力无明显改变,LDH释放和细胞凋亡率略有升高,但差异无统计学意义(P>0.05)。低氧状态下,M+H组较C+H组LDH释放、细胞凋亡率明显降低(P<0.05),细胞活力明显升高(P<0.05);I+H组较C+H组LDH释放、细胞凋亡率明显升高(P<0.05),细胞活力明显降低(P<0.05)。结论 miR-486能够减轻心肌细胞低氧损伤、增强心肌细胞活力、降低细胞凋亡率,对细胞低氧损伤和凋亡有改善作用。

microRNA-486对低氧导致的原代大鼠心肌细胞损伤和凋亡的改善作用

张海涛¹, 韦薇², 齐弘炜³, 袁彪³, 汤楚中⁴

1. 承德医学院附属医院心脏外科, 河北 承德 067000;
2. 承德医学院附属医院血液检验室, 河北 承德 067000;
3. 首都医科大学附属北京同仁医院心脏外科, 北京 100730;
4. 中国人民解放军总医院第六医学中心心脏外科, 北京 100048

收稿日期:2019-09-10出版日期:2020-06-30发布日期:2020-06-15
通讯作者:汤楚中E-mail:tangchuzhong@sohu.com
作者简介:张海涛(1986-),男,主治医师,硕士.
基金资助:国家自然科学基金（81370237）


关键词: microRNA-486, 腺病毒, 低氧, 细胞凋亡
Abstract: Objective To elucidate the effects of microRNA-486 (miR-486) on the hypoxia-induced injury of primary cardiomyocytes. Methods Primary cardiomyocytes from neonatal rats were cultured,and models of myocardial hypoxia-induced injury were constructed by inducing acute ischemia. The expression of miR-486 under conditions of hypoxia was detected using real-time PCR at 6,12,24,36,and 48 hours. The mice were divided into six groups according to the method of transfection of Ad-miR-486 and Ad-miR-486-antago,and comprised the normoxic group (C+N group),normoxic group transfected with Ad-miR-486 (M+N group),normoxic group transfected with Ad-miR-486-antago (I+N group),hypoxic group (C+H group),hypoxic group transfected with Ad-miR-486 (M+H group),and hypoxic group transfected with Ad-miR-486-antago (I+H group). The cell viability,injury,and apoptosis were measured by determining the levels of CCK-8 and lactate dehydrogenase (LDH),and by using flow cytometry. Results The expression of miR-486 at 12,24,and 36 hours was significantly higher than that of the control group. There were no significant changes in cell viability,LDH release,and rate of apoptosis among the normoxic groups. Under the conditions of hypoxia in the M+H group,the amount of LDH released and the rate of apoptosis significantly decreased,while the cell viability significantly increased,than in the C+H group. In the I+H group,the amount of LDH released and the rate of apoptosis were significantly higher,while the cell viability significantly reduced,than in the C+H group. Conclusion miR-486 improved cell injury and apoptosis under conditions of hypoxia by alleviating cell injury,enhancing cell viability,and inhibiting apoptosis.
Key words: microRNA-486, adenovirus, hypoxia, apoptosis
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