透射电镜在糖尿病大鼠肠上皮细胞连接分析中的应用

Application of transmission electron microscopy in the analysis of intestinal epithelial cell connections in diabetic rats

探讨透射电镜技术在糖尿病大鼠肠上皮细胞连接分析中的应用。采用高脂饮食联合链脲佐菌素(STZ)腹腔注射方法建立糖尿病大鼠模型,采用罗氏血糖仪检测大鼠尾血血糖,采用透射电镜观察大鼠回肠上皮细胞连接的病理学改变,采用免疫印迹检测大鼠回肠组织claudin、occludin、ZO-1蛋白表达水平。成功构建糖尿病大鼠模型。糖尿病大鼠肠上皮细胞间紧密连接与正常对照组比较稍感松弛,电子密度有所降低;中间连接、缝隙连接则出现缝隙异常增宽现象。与正常对照组比较,模型组细胞连接相关蛋白claudin、occludin、ZO-1表达水平显著降低。透射电镜技术可应用于糖尿病大鼠肠上皮细胞连接病理形态学分析,对糖尿病大鼠肠黏膜屏障功能评价具有重要意义。

Objective： To investigate the application of transmission electron microscopy （TEM） in the analysis of intestinal epithelial cell connectivity in diabetic rats. Methods： A diabetic rat model was established by high fat diet combined with intraperitoneal injection of STZ. The blood glucose in tail blood of rats was detected by Roche glucose meter. The pathological changes of the ileum epithelial cell connections were observed by transmission electron microscopy. The expression levels of claudin, occludin and ZO-1 in the ileum of rats were detected by Western blotting. Results： The model of diabetic rats was successfully constructed. Compared with the normal control group, the blood glucose value was significantly higher in diabetic rat models, and the blood sugar remained on a high level after 4 weeks of continuous feeding. Compared with the normal control group, diabetic rats showed a little relaxation in intestinal epithelial tight junctions between cells, and decreased electron density. The gap of intermediate junction and gap junction appeared abnormally wide. The expression level of cell junction associated protein claudin, occludin, ZO-1 were significantly lower. Conclusion： Transmission electron microscopy （TEM） can be used for pathological analysis of intestinal epithelial cell junction in diabetic rats, and is of great significance for evaluation of intestinal mucosal barrier function in diabetic rats.t