Ma Meng, Ma Xiaohui, Cui Jie, Guo Yifeng, Tang Xiuqin, Chen Chuanmin, Zhu Ying, Cui Chao, Wang Gang
State Key Laboratory of Microbial Technology, Shandong University, Qingdao, Shandong, China.
Department of Pharmacy, Taian Maternal and Child Health Hospital, Taian, Shandong, China.
Cardiovasc Eng Technol. 2019 Jun;10(2):329-343. doi: 10.1007/s13239-019-00404-7. Epub 2019 Feb 6.
This study aims to investigate the effects and the molecular mechanism of cyclosporin A (CsA) against oxidative stress injury in cultured neonatal rat cardiomyocytes.
Bax/Bcl-2, cl-casp-9/casp-9, cl-casp-3/casp-3, and iNOS/β-actin ratios and p-IκB and IκB levels were analyzed by western blot. IL-1β and TNF-α levels were analyzed by ELISA.
CsA effectively improved the cell viability and reduced the extracellular lactate dehydrogenase release in cardiomyocytes after HO-induced oxidative damage. CsA significantly increased the superoxide dismutase activity, glutathione production, and catalase activity but decreased the malonaldehyde level. CsA treatment considerably reduced the HO-induced intracellular generation of reactive oxygen species, mitochondrial dysfunction, and release of cytochrome c. CsA could act against HO-induced ATP reduction, TCA cycle enzymes, mitochondrial complex I enzyme, and complex V enzyme in cardiomyocytes. CsA significantly decreased the Bax/Bcl-2 ratio, cl-casp-9/casp-9, and cl-casp-3/casp-3 in a concentration-dependent manner. CsA also remarkably reduced the cleaved PARP level and DNA fragmentation. NF-κB was closely related to oxidative stress injury. CsA inhibited NF-κB activation, thereby preventing the upregulation of IL-1β, TNF-α, iNOS, and intracellular NO release.
CsA protected cardiomyocytes against HO-induced cell injury. Hence, CsA may be developed as a candidate drug to prevent or treat myocardial ischemia reperfusion injury.
本研究旨在探讨环孢素A（CsA）对培养的新生大鼠心肌细胞氧化应激损伤的影响及其分子机制。
采用蛋白质免疫印迹法分析Bax/Bcl-2、cl-casp-9/casp-9、cl-casp-3/casp-3和iNOS/β-肌动蛋白的比值以及p-IκB和IκB的水平。采用酶联免疫吸附测定法分析白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）的水平。
在过氧化氢（HO）诱导的氧化损伤后，CsA有效提高了心肌细胞的活力并减少了细胞外乳酸脱氢酶的释放。CsA显著提高了超氧化物歧化酶活性、谷胱甘肽生成量和过氧化氢酶活性，但降低了丙二醛水平。CsA处理显著减少了HO诱导的细胞内活性氧生成、线粒体功能障碍和细胞色素c的释放。CsA可以对抗HO诱导的心肌细胞中ATP减少、三羧酸循环酶、线粒体复合物I酶和复合物V酶的损伤。CsA以浓度依赖的方式显著降低了Bax/Bcl-2比值、cl-casp-9/casp-9和cl-casp-3/casp-3。CsA还显著降低了裂解的聚（ADP-核糖）聚合酶水平和DNA片段化。核因子κB（NF-κB）与氧化应激损伤密切相关。CsA抑制NF-κB的激活，从而阻止IL-1β、TNF-α、诱导型一氧化氮合酶（iNOS）的上调和细胞内一氧化氮的释放。
CsA保护心肌细胞免受HO诱导的细胞损伤。因此，CsA可能被开发成为预防或治疗心肌缺血再灌注损伤的候选药物。
