Objective To observe the effects of the preconditioning of ulinastatin on GES-1 cell injury induced by oxygen and glucose deprivation (OGD). Methods GES-1 cells were cultured in vitro and divided into three groups: normal control group (group N), oxygen and glucose deprivation group (group O), and ulinastatin preconditioning group (group U). The OGD model of GES-1 cells were established by glucose-free medium and three-gas incubator for 6h. Ulinastatin was added to group U 12h before the deprivation of oxygen and glucose. The cell viability and apoptosis were determined by cck-8 and flow cytometry respectively. Western Blot was used to examine the protein expression of Caspase-3 and Cleaved Caspase-3. The TRPV1 mRNA expression was measured by quantitative real-time PCR. Results As compared with group N, the viability of GES-1 was decreased, the apoptotic rate and the expression of Caspase-3 and Cleaved Caspase-3 were increased, and the TRPV1 mRNA expression decreased greatly in group O (P < 0.05). As compared with group O, the aforementioned changes were significantly inhibited in group U. Conclusions Ulinastatin preconditioning could effectively inhibit GES-1 cell injury induced by OGD, which may be related to the inhibition of apoptosis and the upregulation of TRPV1 mRNA expression.%目的:观察乌司他丁(UTI)预处理对氧糖剥夺(OGD)诱导人胃黏膜上皮细胞株(GES-1)细胞损伤的影响.方法:采用GES-1细胞株进行实验,设置正常对照组(N组)、氧糖剥夺组(O组)、乌司他丁预处理组(U组).N组不做任何处理,O组和U组通过三气培养箱和无糖培养基共同作用造成细胞氧糖剥夺损伤,U组在损伤前12 h预先加入乌司他丁处理.损伤处理6 h后,采用CCK-8法测定细胞活力,流式细胞术检测细胞凋亡率,免疫印迹法检测Caspase-3和Cleaved Caspase-3的蛋白表达水平,实时荧光定量PCR法检测细胞内瞬时受体电位香草酸亚型-1(TRPV1)分子mRNA的表达水平.结果:与N组比较,O组细胞存活率显著降低,细胞凋亡率明显升高,Caspase-3和Cleaved Caspase-3表达增加,TRPV1 mRNA表达降低(P<0.05);乌司他丁预处理能显著抑制上述O组的变化,差异有统计学意义.结论:乌司他丁预处理可减轻OGD诱导的GES-1细胞损伤,其机制可能与抑制细胞凋亡以及调控TRPV1的表达有关.
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