亚硒酸钠对糖尿病肾病大鼠肾脏Nephrin表达的影响与意义

The Influence of Sodium Selenite on Nephrin Gene Expression in Kidney of Diabetic Nephropathy Rats and Significance

探讨亚硒酸钠对糖尿病肾病大鼠肾脏Nephrin表达的影响及二者间的关系,从而研究亚硒酸钠和Nephrin在糖尿病肾病中的作用机制.通过链脲佐菌素法及给予高脂饮食诱导模拟大鼠糖尿病肾病模型,实验设空白对照组、糖尿病肾病对照组、亚硒酸钠干预组,亚硒酸钠干预组每日给予亚硒酸钠溶液灌胃,其它组给予等量生理盐水灌胃.灌胃10周后处死大鼠,取血、尿标本测相关生化指标.取肾脏组织戊二醛固定制作切片电镜下观察超微结构改变,取肾脏组织多聚甲醛固定制石蜡切片光镜下观察病理改变和免疫组化定位蛋白表达.取肾脏组织RT-PCR检测Nephrin的mRNA表达、Western Blotting检测nephrin的蛋白表达,分析各组数据的统计差异.结果发现亚硒酸钠干预组大鼠基本状况和生化指标较糖尿病肾病对照组明显改善,光镜和电镜下观察病理改变和超微结构病变较糖尿病肾病对照组明显减轻.免疫组化nephrin蛋白表达着色糖尿病肾病对照组较空白对照组减少,亚硒酸钠干预组较糖尿病肾病对照组着色明显增多.Nephrin mRNA和蛋白表达糖尿病肾病对照组较空白对照组明显降低,而亚硒酸钠干预组较糖尿病肾病对照组升高,但低于空白对照组,差异均有统计学意义(P<0.05).亚硒酸钠明显促进肾脏Nephrin表达,改善了糖尿病肾病,表明亚硒酸钠和Nephrin在防治和延缓糖尿病肾病的发生发展中可能起重要作用.

To investigate the influence of sodium selenite on Nephrin gene expression in kidney of diabetic nephropathy rats and the relation between sodium selenite and Nephrin, so as to study the role mechanism of sodium selenite and Nephrin in diabetic nephropathy. Diabetic nephropathy rat models were established by injecting streptozocin intraperitoneally and feeded high fat diet. Three different groups of the rats were employed： control group, diabetic nephropathy group and selenite-treated group. The selenite-treated group taked selenite solution by intragastric administration every day, and other groups took same dose saline by intragastric administration every day. After 10 weeks, rats were killed, blood and urine sample were got ten to took some biochemical indicators. The kidney was cut and made section by glutaral fixation to observe fine structure changes through electron microscope. The kidney was cut and made paraffin section to observe pathological changes with light microscope and to locate nephrin protein expression with immunohistochemistry analysis. The kidney was cut and the expression of Nephrin mRNA and nephrin protein through RT-PCR and Western blotting were examined. All data was analyzed by statistics methods. The basic conditions and biochemical indicators of selenite-treated group were better than diabetic nephropathy group, and the same to pathological changes and fine structure changes. Immunohistochemistry analysis manifested that the coloration of diabetic nephropathy group paraffin section diminished comparing to the control group, but the coloration of selenite-treated group paraffin section enhanced comparing to the diabetic nephropathy group. The expression of Nephrin mRNA and nephrin protein in diabetic nephropathy group was decreased comparing to the control group, but of which selenite-treated group enhanced comparing to the diabetic nephropathy group, diminishing comparing to the control group yet. All differences were statistically significant （P〈0.05）. Sodium selenite can enhance the expression of Nephrin in kindey of diabetic nephropathy rats, improve diabetic nephropathy, and it is hinted that sodium selenite and Nephrin may play important roles in delaying the process of diabetic nephropathy.