牛磺酸对肾NRK-52E细胞缺氧-复氧损伤的保护作用研究

Protective activity of taurine against ischemia reperfusion injury in renal NRK-52E cells

目的观察牛磺酸对肾NRK-52E细胞缺氧-复氧损伤的保护作用,探讨其相关的作用机制。方法建立缺氧-复氧细胞模型,实验细胞分为四组:正常对照(A)组、缺氧-复氧(B)组、低浓度牛磺酸干预组(C)组、高浓度牛磺酸干预组(D)组。台盼蓝染色计算细胞存活率,Annexin V/PI双染流式细胞仪检测细胞凋亡的发生,Western blot检测蛋白表达的变化。结果 A、B、C、D组细胞的存活率分别为(96.2±3.4)%、(58.7±6.2)%、(66.9±6.7)%、(75.8±7.1)%,各组之间存在显著性差异(P<0.05)。A、B、C、D组细胞的存活率分别为(3.6±1.4)%、(36.8±5.2)%、(28.5±4.8)%、(17.6±4.2)%,各组之间存在显著性差异(P<0.05)。缺氧-复氧上调了肾NRK-52E细胞GRP78和Caspase-12蛋白表达,牛磺酸可以显著抑制缺氧-复氧处理所刺激的GRP78和Caspase-12蛋白表达的上调。结论牛磺酸可以抑制肾细胞的缺氧-复氧损伤,下调GRP78和Caspase-12蛋白抑制内质网应激可能是其主要的作用机制。

[ Objective ] To study the protective activity of taurine against ischemia reperfusion injury in renal NRK-52E ceils and its action mechanism. [ Methods ] The hypoxia-reoxygenation cell model was established. The experimental cells were divided into four groups： normal control group（A）, hypoxia-reoxygenation group（B）, low con- centrations of taurine intervention group （C）, high concentrations of taurine intervention group （D） group. Trypan blue was used to detect cell viability. The apoptosis was determined by Annexin V-FITC/PI staining and flow cytometry analysis. The protein expression was detected by Western blot analysis. [Results] The cells survival rates were （96.2±3.4）%, （58.7±6.2）%, （66.9±6.7）%, （75.8±7.1）% in A, B, C, D group. There was a significant difference among the groups（P 〈0.05）. The percent of apoptosis was （3.6±1.4）%, （36.8±5.2）%, （28.5±4.8）%,（ 17.6±4.2）% in A, B, C, D group. There was a significant difference among the groups （P 〈0.05）. The expression of GRP78 and Caspase-12 protein was unregulated in kidney NRK-52E cells by hypoxia-reoxygenation. Taurine significantly inhibited the hy- poxia - reoxygenation-stimulated up-regulation of GRP78 and Caspase-12. [ Conclusions ] Taurine inhibited hy- poxia-reoxygenation injury in renal cell, and which was mainly related to the downregulation of GRP78 and Cas- pase-12 protein.