Zhang Qin, Dong Xin-Wei, Xia Jia-Ying, Xu Ke-Ying, Xu Zhe-Rong
Department of Geriatrics, First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China (mainland).
Department of Pharmacology, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China (mainland).
Med Sci Monit. 2017 May 4;23:2127-2136. doi: 10.12659/msm.901361.
BACKGROUND Obestatin, primarily recognized as a peptide within the gastrointestinal system, has been shown to benefit the cardiovascular system. We designed this experiment to study the protective role and underlying mechanism of obestatin against ischemia-reperfusion(I/R) injury in myocardial cells. MATERIAL AND METHODS In an In vivo experiment, LAD was ligated for 0.5 h and then opened for reperfusion with obestatin for 24 h. Then, the infarction area was shown with TTC staining, and inflammation factors in serum were analyzed by qRT-PCR. In primary cultured cardiomyocytes, we measured the level of LDH, MDA, GSH, and SOD. Finally, we assessed cells apoptosis using flow cytometry and detected the concentrations of caspase-3, Bax, and Bcl-2 using Western blot analysis. RESULTS TTC staining showed that in the 3 obestatin groups, the infarct area became smaller with the increase of obestatin concentration. Obestatin also inhibited LDH expression in rat serum and decreased mRNA levels of TNF-α, IL-6, ICAM-1, and iNOS in rat cardiomyocytes after reperfusion. In primary cultured cardiomyocytes, obestatin decreased LDH content and increased GSH level after I/R injury. Obestatin was also found to antagonize the apoptosis of cardiomyocytes in a dose-dependent manner. Western blot analysis showed that obestatin downregulated the expression of caspase-3 and Bax and upregulated the expression of Bcl-2. CONCLUSIONS Obestatin can protect cardiomyocyte from I/R-induced injury in vitro and in vivo. This beneficial effect is closely related with its properties of anti-inflammation, anti-cytotoxicity, and anti-apoptosis. The protective effect of obestatin might be associated with activation of Bcl-2 and inhibition of caspase-3 and Bax.
背景 肥胖抑制素最初被认为是一种胃肠道系统中的肽，已被证明对心血管系统有益。我们设计了本实验来研究肥胖抑制素对心肌细胞缺血再灌注（I/R）损伤的保护作用及其潜在机制。材料与方法 在体内实验中，结扎左冠状动脉前降支（LAD）0.5小时，然后开放再灌注，同时给予肥胖抑制素24小时。然后，用TTC染色显示梗死面积，并用qRT-PCR分析血清中的炎症因子。在原代培养的心肌细胞中，我们测量了乳酸脱氢酶（LDH）、丙二醛（MDA）、谷胱甘肽（GSH）和超氧化物歧化酶（SOD）的水平。最后，我们用流式细胞术评估细胞凋亡，并通过蛋白质免疫印迹分析检测半胱天冬酶-3（caspase-3）、促凋亡蛋白（Bax）和抗凋亡蛋白（Bcl-2）的浓度。结果 TTC染色显示，在3个肥胖抑制素组中，随着肥胖抑制素浓度的增加，梗死面积变小。肥胖抑制素还抑制大鼠血清中LDH的表达，并降低再灌注后大鼠心肌细胞中肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）、细胞间黏附分子-1（ICAM-1）和诱导型一氧化氮合酶（iNOS）的mRNA水平。在原代培养的心肌细胞中，肥胖抑制素降低了I/R损伤后的LDH含量并提高了GSH水平。还发现肥胖抑制素以剂量依赖的方式拮抗心肌细胞的凋亡。蛋白质免疫印迹分析显示，肥胖抑制素下调caspase-3和Bax的表达，并上调Bcl-2的表达。结论 肥胖抑制素在体外和体内均可保护心肌细胞免受I/R诱导的损伤。这种有益作用与其抗炎、抗细胞毒性和抗凋亡特性密切相关。肥胖抑制素的保护作用可能与激活Bcl-2以及抑制caspase-3和Bax有关。
