miR-34a在缺氧复氧介导心肌细胞凋亡中的作用以及当归补血汤的干预效果  

Study on the role of miR-34a in cardiomyocyte apoptosis induced by reoxygenation after hypoxia and the intervention of Danggui Buxue Decoction

作　　者：范智文[1] 赵一俏[2] 凌龙[1] 席玉胜[1] 李铭[1] 靳文[2] 崔同涛 徐基瑛 

机构地区：[1]中国人民解放军第四二一医院心内科,广东广州510318 [2]广东省第二人民医院心内科,广东广州510318 [3]广东省人民医院(广东省医学科学院)心内科,广东广州510080

出　　处：《广东医学》2017年第22期3401-3404,共4页Guangdong Medical Journal

基　　金：广东省自然科学基金资助项目(编号:2016A030313853);广东省中医药局科研课题(编号:20151041);中国人民解放军第四二一医院医学创新项目(编号:201604)

摘　　要：目的探讨miR-34a在缺氧复氧介导心肌细胞凋亡中的作用以及当归补血汤的干预效果。方法培养乳鼠心室肌细胞,制备缺氧复氧模型,并分为对照组、miR-34a抑制组以及当归补血汤低、中、高剂量含药血清组。细胞计数试剂盒-8法检测各组心肌细胞存活率、流式细胞术检测心肌细胞凋亡率,定量PCR法检测miR-34a的表达,Western blot检测Bcl-2的表达。结果对照组、当归补血汤低、中、高剂量含药血清组以及miR-34a抑制组的凋亡细胞比例分别为(35.00±4.69)%、(33.00±5.55)%、(28.83±4.49)%、(25.33±5.43)%、(23.17±5.04)%;相对于对照组,当归补血汤低、中、高剂量含药血清组以及miR-34a抑制组的心肌细胞活力分别为1.290±0.130、1.822±0.123、2.082±0.149、2.238±0.247,miR-34a表达分别为0.877±0.071、0.705±0.045、0.580±0.077、0.453±0.076。与对照组相比,miR-34a抑制组、当归补血汤高、中、低剂量含药血清组的心肌细胞存活率明显升高(P<0.05),心肌细胞miR-34a表达降低(P<0.05),Bcl-2蛋白表达升高(P<0.05)。与对照组和当归补血汤低剂量含药血清组相比,当归补血汤高、中剂量含药血清组心肌细胞凋亡明显减少(P<0.05)。结论当归补血汤能够有效抑制心肌细胞凋亡,与抑制miR-34a表达密切有关。Objective To investigate the role of miR - 34a in hypoxia/reoxygenation - mediated cardiomyocyte apoptosis and the effect of Danggui Buxue Decoction. Methods Cultured neonatal rat ventricular myocytes were prepared by hypoxia and reoxygenation model, and divided into control group, miR -34a inhibitory group and low - , medium - and high - dose Danggui Buxue Decoction groups. Cell counting kit - 8 method was used to detect the viability of cardio- myoeytes. Flow cytometry was used to detect the apoptosis of cardiomyocytes. The expression of miR - 34a was assessed by quantitative PCR. The expression of Bcl -2 was assessed by Western blot. Results The apoptosis ratio in control group, low - , medium - and high - dose Danggui Buxue Decoction groups and miR - 34a inhibitory group were （ 35.00± 4. 69） %, （33. 00 ± 5.55 ） %, （28.83± 4.49 ） %, （25.33 ±5.43 ） % and （23.17 ± 5.04） %, respectively. In contrast to control group, the cell survive ratios in low -, medium - and high - dose Danggui Buxue Decoction groups and miR - 34a inhibitory group were 1. 290 ±0. 130, 1. 822 ±0. 123, 2. 082 ±0. 149 and 2. 238 ±0. 247, respectively. In contrast to control group, the miR - 34a expression in low - , medium - and high - dose Danggui Buxue Decoction groups and miR -34a inhibitory group were 0. 877±0. 071,0. 705 ±0. 045, 0. 580 ±0. 077 and 0. 453 ±0. 076, respectively. Compared with control group, the viability of cardiomyocytes and the expression of Bcl - 2 protein were significantly improved, and the expression of miR - 34a was decreased in miR - 34a inhibitory group and low - , medium - and high - dose Dan- ggui Buxue Decoction groups （ all P 〈 0. 05 ）. Compared with control group and low - dose Danggui Bnxue Decoction group, the apoptosis of cardiomyocytes in medium - and high - dose Danggui Buxue Decoction group were decreased （P 〈0. 05）. Conclusion Danggui Buxue Decoction can effectively inhibit the apoptosis of cardiomyocytes, which is closely related to the inhibition of miR - 3

关 键 词：心肌细胞 细胞凋亡 当归补血汤 MIR-34A 

分 类 号：R542.2[医药卫生—心血管疾病]