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目的:考察酒石酸对新生大鼠原代心肌细胞缺氧复氧损伤的保护作用。方法:大鼠乳鼠进行心肌细胞原代培养,MTT法检测心肌细胞的活力。复制心肌细胞缺氧复氧损伤模型,考察酒石酸对心肌细胞缺氧复氧损伤所致的LDH漏出的影响,并进一步分别采用流式细胞术及Western blot考察酒石酸对心肌细胞凋亡,cleaved-caspase 3,Akt及P-Akt的影响。结果:(1)酒石酸31.25～500μg/ml范围对原代心肌细胞活力不产生影响。酒石酸400μg/ml均可显著降低心肌细胞缺氧复氧损伤后LDH漏出率,而200、100、50μg/ml浓度时作用均不明显;(2)酒石酸400、200μg/ml有抑制心肌细胞缺氧复氧损伤所致凋亡百分数增加的趋势;(3)酒石酸200μg/ml均可显著抑制心肌细胞缺氧复氧所致的cleaved-caspase 3蛋白表达增加,而酒石酸400μg/ml有降低cleaved-caspase 3蛋白表达的趋势,无统计学差异;(4)酒石酸400μg/ml可显著增加心肌细胞的p-Akt蛋白表达,而酒石酸200μg/ml对p-Akt蛋白表达未见明显影响。结论:酒石酸可以抑制心肌细胞缺氧复氧损伤所致的坏死和凋亡,可能是通过激活Akt的磷酸化而实现的。
Objective: To investigate the protective effect of tartaric acid on hypoxia-reoxygenation injury in neonatal rat cardiomyocytes. Methods: Primary rat cardiomyocytes were cultured in neonatal rat and the activity of cardiomyocytes was measured by MTT assay. To investigate the effects of tartaric acid on LDH leakage induced by anoxia-reoxygenation injury in cardiomyocytes and the effects of tartaric acid on the apoptosis of cardiomyocytes, cleaved-caspase 3, Akt and P-Akt. Results: (1) The range of 31.25-500 μg / ml tartaric acid did not affect the viability of primary cardiomyocytes. Tartaric acid 400μg / ml can significantly reduce LDH leakage rate of myocardial cells after hypoxia-reoxygenation injury, and 200,100,50μg / ml concentration of the role was not obvious; (2) 400,200μg / ml tartaric acid can inhibit myocardial cell hypoxia (3) Tartaric acid at 200μg / ml could significantly inhibit the expression of cleaved-caspase 3 protein induced by hypoxia-reoxygenation in cardiomyocytes, whereas tartaric acid at 400μg / ml could reduce cleaved-caspase (4) 400μg / ml tartaric acid could significantly increase the expression of p-Akt protein in cardiomyocytes, while no significant effect of p-Akt protein expression was found at 200μg / ml tartaric acid. Conclusion: Tartaric acid can inhibit necrosis and apoptosis induced by hypoxia-reoxygenation injury in cardiomyocytes, probably through the activation of Akt phosphorylation.