高尿酸通过转运蛋白引起大鼠小肠黏膜上皮细胞炎性反应

Inflammatory Response of Intestinal Mucosa Epithelial Cells Induced by High Uric Acid through TSPO Transporter Protein in Rats

研究转运蛋白(TSPO)在高尿酸引起肠上皮细胞炎症反应中的作用及机制。高尿酸体外处理肠黏膜上皮细胞,通过荧光定量PCR方法检测转运蛋白,ATP结合膜转运蛋白(ABCG2)、闭锁蛋白Occludin、上皮紧密连接相关蛋白-1(ZO-1)和IL-1β等基因表达水平,采用Western blot检测NF-κB信号通路中p65和IL-1β表达水平,采用试剂盒检测线粒体活性氧的变化,从分子水平及氧化应激反应的角度研究尿酸诱导的肠上皮细胞的炎症反应机制。最后研究高尿酸血症小鼠和正常小鼠的肠渗透性变化。结果表明,与对照组相比,高尿酸处理的肠上皮细胞TSPO、ABCG2、IL-1β等基因的mRNA的表达量显著上升(P<0.05),紧密连接相关蛋白ZO-1、Occludin表达量降低(P<0.05),p65蛋白和IL-1β的蛋白表达升高(P<0.05),活性氧(ROS)产生增高(P<0.05);高尿酸小鼠的肠渗透性明显高于正常小鼠(P<0.05)。说明高尿酸可以通过转运蛋白引起肠上皮细胞炎症反应,增加肠渗透性。

To study the roles and mechanism of TSPO in intestinal epithelial inflammatory response induced by high uric acid,mitochondrial membrane translocation protein(TSPO),ATP binding membrane transporter(ABCG2),Occludin,epithelial tight junction associated protein-1(ZO-1)and IL-1βexpression were detected by quantitative PCR.The levels of p65 and IL-1βwas detected using Western blotting.The mitochondrial ROS was detected using DCFH-DA method.We explored intestinal epithelial inflammatory response induced by high uric acid in the perspective of molecular mechanism and oxidative stress response.Finally,the changes of intestinal permeability in hyperuricemic mice and wild-type mice were studied in vivo,respectively.Compared with the control group,the mRNA expression of TSPO,ABCG2,and IL-1βwere increased significantly(P<0.05),whereas Occludin and ZO-1 were decreased(P<0.05).Both of the protein expression of p65 and IL-1βincreased significantly,and the production of ROS increased significantly(P<0.05).The intestinal permeability of hyperuricemia mice was significantly higher than that of wild mice(P<0.05).High uric acid can cause the inflammatory response of the intestinal epithelial cells through TSPO transporter and increase intestinal permeability.