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目的探讨黄连素(BR)对大鼠H9c2心肌细胞缺氧/复氧(H/R)损伤后凋亡的影响及其作用机制。方法将培养的H9c2心肌细胞随机分为:正常对照组(NC组)、单纯药物组(BR组)、缺氧/复氧组(H/R组)、药物低剂量LBR组(1.5×10-6mol/L)及高剂量HBR组(1.5×10-4mol/L)。处理结束后,用MTT比色法检测H9c2心肌细胞的活力,用Hoechst33258染色检测细胞核形态的变化,用Western blot法检测Nrf2,Keap1蛋白的表达。结果与NC组比较,单纯BR对H9c2心肌细胞无影响。与H/R组比较,低、高剂量BR组细胞的活力明显上升(65.2±1.6%;82.3±1.4%)(P<0.01),心肌细胞的凋亡率明显减少(P<0.05),Western blot的结果显示,BR可明显促进抗氧化相关蛋白Nrf2表达,抑制Keap1的表达。结论 BR对H9c2心肌细胞H/R损伤后的凋亡具有显著的抑制作用,可能与其促进抗氧化核转录因子Nrf2表达有关。
Objective To investigate the effect and mechanism of berberine (Bcl-2) on the apoptosis of rat H9c2 cardiomyocytes after hypoxia / reoxygenation (H / R) injury. Methods The cultured H9c2 cardiomyocytes were randomly divided into normal control group (NC group), pure drug group (BR group), hypoxia / reoxygenation group (H / R group) and low dose LBR group (1.5 × 10- 6mol / L) and high dose HBR group (1.5 × 10-4mol / L). After treatment, the viability of H9c2 myocardial cells was detected by MTT colorimetric assay. The changes of nuclear morphology were detected by Hoechst33258 staining. The expression of Nrf2 and Keap1 protein was detected by Western blot. Results Compared with NC group, BR alone had no effect on H9c2 cardiomyocytes. Compared with H / R group, the viability of cells in low and high doses of BR was significantly increased (65.2 ± 1.6% vs 82.3 ± 1.4%, P <0.01), and the apoptosis rate of cardiomyocytes was significantly decreased (P <0.05) The results of blot showed that BR could obviously promote the expression of Nrf2, and inhibit the expression of Keap1. Conclusion BR has a significant inhibitory effect on H9c2 cardiomyocytes apoptosis after H / R injury, which may be related to the promotion of the expression of Nrf2, an antioxidant nuclear factor.