养阴活血方药对糖尿病大鼠肾脏的保护作用及机制探讨

Protective effect of Nourishingyin and promotingblood flow recipe on kidney of diabetic rat

目的:探讨养阴活血方药(nourishing yin and promoting blood flow recipe,NYPBR)对糖尿病大鼠肾脏的保护作用机制。方法:SD大鼠随机分为正常组、模型组和NYPBR组,后两组大鼠腹腔注射链脲佐菌素复制糖尿病模型。正常组和模型组自由饮水。NYPBR组按人与动物间体表面积折算的等效剂量比值灌喂给药(含生药1 g·mL^(-1)),每只大鼠3 mL·d^(-1)。10周后,RT-PCR检测肾皮质诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)mRNA表达,Western blot检测iNOS蛋白含量,免疫组化检测过氧亚硝基阴离子(peroxynitrite,ONOO^-)特异性标志物-硝基酪氨酸(nitrotyrosine,NT)的生成。光镜观察大鼠肾皮质形态学变化。检测各组大鼠肾皮质超氧化物歧化酶(superoxide dismutase,SOD)活性和丙二醛(malondialdehyde,MDA)含量及血糖、24 h尿蛋白定量和肌酐清除率。结果:与正常组相比,模型组肾皮质iNOS mRNA表达0.90±0.10和蛋白量含量43.00±6.08增加、NT生成87.23±5.94明显增加,与肾皮质病理改变及肾功能减退呈一致性变化。NYPBR可显著改善上述变化。结论:NYPBR可显著减轻糖尿病大鼠的肾脏损伤,此作用可能与降低iNOS mRNA表达和蛋白含量、减少ONOO^-的生成有关。

Objective： To investigate the protective effects of Nourishingyin and Promotingblood flow recipe （NYPBR） on the kidney of diabetic rat. Method： SD rats were divided into 3 groups at random ： control group, diabetes group and NYPBR group. The latter two groups were injected intraperitoneally with streptozotocin to induce diabetes model. Rats in NYPBR group were fed NYPBR solution （3 g·d^-1）, with dose equivalent to the clinical use in the patients. Rats in the other groups were fed equivalent water. 10 weeks after diabetes was induced, the inducible nitric oxide synthase （iNOS） mRNA expression in the renal cortex was detected by RT-PCR, and its protein content by Western blotting. Immunohistochemistry was used to detect the formation of nitrotyrosine （NT）, a specific marker of peroxynitrite （ONOO^- ）. The morphological changes of renal cortex were observed under optical microscope. Superoxide dismutase （SOD） and malondialdehyde （MDA） in renal cortex, blood glucose, 24 h urine protein content and creatinine clear-ance rate in different groups were detected. Result： Compared with control group, the iNOS mRNA expression （0.90±0. 10） and its protein content （43.00±6.08）, and NT content （87.23±5.94） increased significantly in diabetes group, in accord with the patho- logical changes of renal cortex and renal dysfunction. NYPBR can attenuate the pathological alterations. Conclusion： NYPBR could decrease iNOS mRNA expression and its protein content, and reducing the overformation of ONOO^-, thus protecting the kidney of diabetic rat from injury.