体外循环再灌注血培养诱导肾小管细胞凋亡及乌司他丁的保护作用研究

Protective Effect of Ulinastatin on Apoptosis of Cultured Human Proximal Tubular Cells Induced by in Vitro Cardiopulmonary Bypass Serum

目的探索乌司他丁对体外循环血清致人肾小管上皮细胞损伤的保护作用。方法以人近曲肾小管上皮细胞HK-2为研究对象,根据是否应用体外循环血清将细胞分为空白组:不含人血清的培养基;对照组:含正常人血清的培养基;试验组:含体外循环血清的培养基,进行缺氧复氧培养,通过流式细胞仪检测肾小管细胞凋亡情况,免疫组化检测caspase-3蛋白表达;乌司他丁组:以改良的缺血再灌注方式处理肾小管上皮细胞的过程中,施加乌司他丁干预。结果流式细胞仪检测结果显示,体外循环血清处理后,4组肾小管上皮细胞HK-2凋亡率比较,差异有统计学意义(P<0.05);其中试验组较空白组、对照组、乌司他丁组均明显升高,差异有统计学意义(P=0.00)。4组caspase-3蛋白积分分光光度值(IOD)值比较差异有统计学意义(P<0.05);其中试验组较空白组、对照组、乌司他丁组均明显升高,差异有统计学意义(P=0.00)。结论乌司他丁可通过抑制caspase-3激活,减轻体外循环缺血再灌注血清对HK-2损伤作用。

Objective To investigate the protective effect of Ulinastatin on human proximal tubular cells （ HK - 2） in- jured by in vitro cardiopulmanary bypass （CPB） serum through improved ischemia reperfusion model. Methods HK - 2 cells were used as target cells. According to whether using CPB serum or not, the ceils were divided into blank group （ human serum - free medium）, control group （medium containing normal human serum）, experiment group （medium containing CPB serum, given hypoxia and reoxygenation culture, and the apoptosis rate of HK - 2 cells was detected by flow cytome/er and the protein ex- pression was detected by immunohistochemical technique） and Ulinastatin group （ during the process of using improved ischemia reperfusion method treating HK -2 cells, Ulinastatin was used to intervene the treatment course） . Results The results of flow cytometer showed that after the treatment of CPB serum, the apoptosis rate of HK - 2 cells between the four groups showed statisti- cally significant differences （P 〈 0. 05） . Compared with blank group, control group and Ulinastatin group, the apoptosis rate of experiment group was significantly higher （P = 0. 00） . The IOD value of caspase - 3 protein between the four groups showed sta- tistically significant difference （P 〈 0. 05） . Compared with blank group, control group and Ulinastatin group, the IOD value of experiment group was significantly higher （ P = 0. 00） . ConcluSion Ulinastatin can alleviate the damage on HK - 2 cells by CPB serum through inhibiting the activation of caspase - 3.