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目的研究氨氯地平抑制缺血再灌注诱导的心肌细胞损伤。方法 30只Wistar大鼠随机分为假手术组、模型组、实验组,每组10只。模型组及实验组,通过结扎大鼠左冠状动脉前降支,制作心肌缺血再灌注(MIRI)模型。假手术组及模型组,于术后腹腔注射0.9%NaCl 5 m L;实验组,腹腔注射2 mg·kg~(-1)氨氯地平5 m L,连续灌胃给药7 d。以流式双染法检测心肌细胞凋亡情况,用免疫印迹法分析心肌组织中B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)蛋白表达及磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)通路的激活状况,以分光光度法检测天冬氨酸蛋白水解酶3(Caspase 3)活性。结果与假手术组的早期凋亡率(2.34±0.35)%及晚期细胞凋亡率(3.58±0.39)%比较,模型组早期凋亡率(15.69±1.14)%及晚期细胞凋亡率(24.74±2.56)%显著提高(P<0.05)。假手术组的Bax表达量为(0.18±0.01)、Caspase 3活性为(1.00±0.10),模型组的Bax表达量(0.62±0.06)及Caspase 3活性(3.98±0.18)显著提高(P<0.05);假手术组的Bcl-2表达量为(0.99±0.10)、PI3K表达量为(0.89±0.06),模型组的Bcl-2表达量为(0.14±0.01)及PI3K表达量为(0.18±0.01)显著下调(P<0.05);假手术组的AKT磷酸化水平为(0.95±0.10),模型组的AKT磷酸化水平为(0.13±0.01)显著降低(P<0.05)。与模型组比较,实验组的早期凋亡率(5.23±0.13)%,晚期细胞凋亡率(8.09±0.35)%;实验组的Caspase 3活性(1.47±0.14),实验组能扭转此变化(P<0.05)。结论氨氯地平通过激活PI3K/AKT信号通路能抑制缺血再灌注诱导的心肌细胞损伤。
Aim To study the effect of amlodipine on myocardial cell injury induced by ischemia-reperfusion. Methods Thirty Wistar rats were randomly divided into sham operation group, model group and experimental group, with 10 rats in each group. Model group and experimental group. Myocardial ischemia-reperfusion (MIRI) model was made by ligating left anterior descending coronary artery in rats. Sham-operation group and model group. After intraperitoneal injection of 0.9% NaCl 5 m L, the experimental group was injected intraperitoneally with 2 mg · kg -1 amlodipine 5 m L for 7 days. The apoptosis of cardiomyocytes was detected by flow cytometry. The expression of Bcl-2, Bcl-2 and Bax in myocardium was detected by Western blotting and the expression of phosphatidylinositol 3 Activation of the kinase (PI3K) / protein kinase B (AKT) pathway was measured spectrophotometrically for Caspase 3 activity. Results Compared with the sham operation group, the early apoptosis rate (2.34 ± 0.35)% and the late apoptosis rate (3.58 ± 0.39)%, the early apoptosis rate (15.69 ± 1.14)% and the late apoptosis rate (24.74 ± 2.56)% (P <0.05). The Bax expression in the sham operation group was significantly higher than that in the sham operation group (0.18 ± 0.01), Caspase 3 activity (1.00 ± 0.10), Bax expression in the model group (0.62 ± 0.06) and Caspase 3 activity (3.98 ± 0.18) (0.99 ± 0.10), PI3K (0.89 ± 0.06), the expression of Bcl-2 in the model group (0.14 ± 0.01) and the expression of PI3K in the sham-operated group 0.01). AKT phosphorylation was (0.95 ± 0.10) in sham operation group and AKT phosphorylation level was significantly decreased (0.13 ± 0.01) in model group (P <0.05). Compared with the model group, the early apoptosis rate (5.23 ± 0.13)%, the late apoptosis rate (8.09 ± 0.35)% in the experimental group and the Caspase 3 activity in the experimental group (1.47 ± 0.14) P <0.05). Conclusion Amlodipine can inhibit myocardial cell injury induced by ischemia-reperfusion through activation of PI3K / AKT signaling pathway.