摘要
目的 研究在高糖刺激下H9c2心肌细胞凋亡及凋亡过程中miR-34a(微小核糖核酸-34a的调控作用。方法 将H9c2心肌细胞按葡萄糖浓度不同分为两组(共12孔),使用高浓度葡萄糖(30mmol/L)刺激的观察组(6孔)和低浓度葡萄糖(5 mmol/L)培养的对照组(6孔),利用TUNEL法和流式细胞仪检测H9c2心肌细胞的凋亡情况,miR-34a等相关基因表达采用荧光实时定量PCR测定,通过Western Blot检测B淋巴细胞瘤-2基因(Bcl-2)和半胱氨酸蛋白酶-3(Caspase-3)的表达。结果使用TUNEL法检测,染色计数后计算心肌细胞凋亡率,观察组心肌细胞凋亡率明显高于对照组(P<0.05);使用流式细胞仪检测,观察组心肌细胞凋亡率,明显高于对照组[(9.97±0.32)%vs.(5.94±0.03)%,(P<0.05)];观察组H9c2心肌细胞miR-34a的表达量明显高于对照组,[(2.49±0.41)vs.(0.94±0.06),P<0.05];观察组Bcl-2 mRNA的表达低于对照组[(0.071±0.003)vs.(0.401±0.003),P<0.05];观察组Caspase-3 mRNA的表达高于对照组[(0.602±0.015)vs.(0.326±0.005),P<0.05]。结论 高糖刺激可促使H9c2心肌细胞凋亡,整个过程与miR-34a的高表达密切相关,可在临床为糖尿病心肌病的预防与治疗上提供新的思路。
Objective To study the regulating effect of microRNA-34a(miR-34a) in apoptosis course of H9c2 cardiomyocytes stimulated by high glucose.Methods H9c2 cardiomyocytes(12 wells in total) were divided into observation group stimulated by high concentration of glucose(30 mmol/L,6 wells) and control group stimulated by low concentration of glucose(5 mmol/L,6 wells).The apoptosis of H9c2 cardiomyocytes was detected by using TUNEL method and flow cytometer,and expressions of genes related to miR-34a were detected by using qRT-PCR.The expressions of B-cell lymphoma-2(Bcl-2) and Caspase-3 were detected by using Western blotting assay.Results The results of TUNEL detection showed that the apoptosis rate of H9c2 cardiomyocytes was significantly higher in observation group than that in control group(P<0.05).The results of flow cytometer detection showed that the apoptosis rate of H9c2 cardiomyocytes was significantly higher in observation group than that in control group [(9.97±0.32)% vs.(5.94±0.03)%,(P<0.05)].The expression of miR-34a in H9c2 cardiomyocytes was significantly higher in observation group than that in control group [(2.49±0.41) vs.(0.94±0.06),P<0.05].The expression of Bcl-2 mRNA was lower in observation group than that in control group [(0.071±0.003) vs.(0.401±0.003),P<0.05].The expression of Caspase-3 mRNA was higher in observation group than that in control group [(0.602±0.015) vs.(0.326±0.005),P<0.05].Conclusion High glucose stimulation can promote the apoptosis of H9c2 cardiomyocyte,which is closely related to the high expression of miR-34a in the whole process.This study can provide new ideas for the prevention and treatment of diabetic cardiomyopathy in clinical practice.
作者
黄仲略
黄菲菲
高翔
Huang Zhonglue;Huang Feifei;Gao Xiang(Department of Cardiology,Xiehe Jiangbei Hospital,Wuhan 430100,China;不详)
出处
《中国循证心血管医学杂志》
2022年第4期480-483,共4页
Chinese Journal of Evidence-Based Cardiovascular Medicine
