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-myocytes.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.Com-pared 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 Buxue 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 -34a expression.%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.
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