Liaoning Provincial Key Laboratory of Zoonosis, College of Animal Science & Veterinary Medicine, Shenyang Agricultural University, Shenyang, Liaoning, People's Republic of China.
Adv Exp Med Biol. 2019;1155:451-462. doi: 10.1007/978-981-13-8023-5_42.
Objective To determine whether taurine has protective effects on chicken myocardial apoptosis induced by hypoxic condition through inhibiting calpain-1 derived mitochondrial apoptotic pathway. Methods Chicken primary embryonic myocardial cells were isolated and cultured at 37 °C under a 5% CO atmosphere. Firstly the optimum concentration of taurine or PD150606 was chosen by detecting the cell viability. Chicken cardiomyocytes were cultured in 95% N-5% CO atmosphere for 12 h to produce hypoxic conditions. Before hypoxic treatment, 10 mM taurine and 10 uM PD150606 (a specific calpains inhibitor) were added separately or together. The cell apoptosis was detected by acridine orange/ethidium bromide (AO/EB) double staining. Western blotting was used to determine the protein expressions of calpain-1, cytochrome c, Bcl-2, procaspase-9 and procaspase-3 in the cardiomyocytes. Results Taurine administration effectively attenuated the myocardial apoptosis under hypoxic condition, reduced the calpain-1 protein level. In addition, pre-treated taurine could up-regulate the protein expressions of Bcl-2 and procaspase-3 in hypoxic myocardial cells, down-regulate protein expression levels of cytochrome c and procaspase-9. Moreover, taurine exhibited same inhibition effect as PD150606 on the cell apoptosis and proteins express under hypoxic condition. Conclusions Taurine could attenuate the chicken cardiomyocyte apoptosis impaired by hypoxia through inhibiting calpian-1-derived mitochondrial apoptotic pathway in vitro.
目的 通过抑制钙蛋白酶-1 衍生的线粒体凋亡途径,研究牛磺酸对缺氧诱导的鸡心肌细胞凋亡是否具有保护作用。方法 分离培养鸡原代胚胎心肌细胞,37℃、5% CO2 条件下培养。首先通过检测细胞活力选择牛磺酸或 PD150606 的最佳浓度。将鸡心肌细胞在 95% N2-5% CO2 条件下培养 12h 以产生缺氧条件。在缺氧处理前,分别或同时加入 10mM 牛磺酸和 10uM PD150606(一种特定的钙蛋白酶抑制剂)。吖啶橙/溴化乙锭(AO/EB)双重染色检测细胞凋亡。Western blot 法检测心肌细胞中钙蛋白酶-1、细胞色素 c、Bcl-2、procaspase-9 和 procaspase-3 的蛋白表达。结果 牛磺酸处理可有效减轻缺氧状态下的心肌细胞凋亡,降低钙蛋白酶-1 蛋白水平。此外,牛磺酸预处理可上调缺氧心肌细胞中 Bcl-2 和 procaspase-3 的蛋白表达,下调细胞色素 c 和 procaspase-9 的蛋白表达水平。此外,牛磺酸对缺氧条件下细胞凋亡和蛋白表达的抑制作用与 PD150606 相同。结论 牛磺酸可通过抑制钙蛋白酶-1 衍生的线粒体凋亡途径,减轻体外缺氧诱导的鸡心肌细胞凋亡。