养阴活血方药对糖尿病大鼠主动脉内皮损伤的保护作用

The Protective Effect of Nourishing Yin and Promoting Blood Flow Recipe on the Aortic Endothelium Injury of Diabetic Rats

目的:探讨养阴活血方药(Nourishing Yin and Promoting Blood FlowRecipe,NYPBR)对糖尿病大鼠主动脉内皮损伤的保护作用机制。方法:采用链脲佐菌素制备糖尿病大鼠模型,随机分为正常对照组、模型组和NYPBR组。正常对照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灌胃给药(含生药1 g/mL),每只大鼠3 mL/d。13 w后,RT-PCR检测主动脉诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)mRNA表达,Western blotting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、扫描电镜观察大鼠主动脉内皮形态学变化。并测定大鼠血浆中内皮素-1(endothelin-1,ET-1)、血栓素B2(thromboxane B2,TXB2)以及血清SOD活性、MDA含量等生化指标的变化。结果:与正常对照组比较,模型组大鼠主动脉iNOS mRNA表达及iNOS蛋白含量增加,NT生成明显增加,与主动脉内皮病理变化呈一致性。NYPBR可显著改善上述变化。结论:NYPBR可显著减轻糖尿病大鼠的主动脉内皮损伤,此作用可能与降低iNOS mRNA表达及iNOS蛋白含量、减少ONOO-的生成有关。

Objective：To investigate the protective effects of Nourishing Yin and Promoting Blood Flow Recipe （NYPBR） on aortic endothelium injury of diabetic rats. Methods：The SD rats were divided into 3 groups randomly：control group, modle group and NYPBR group. The latter two groups were injected intraperitoneally with streptozotocin to establish diabetes model. Rats in NYPBR group were fed NYPBR solution（3 mL/d） ,with dose equivalent to the clinical use in the patients. Rats in the other groups were administrated equivalent water. 13 weeks after diabetes was induced, the inducible nitric oxide synthase （iNOS） mRNA expressionin the aorta was detected by RT-PCR, iNOS protein content was determined by Western blotting and immunohistochemistry was used to detect the formation of nitrotyrosine （ NT）, a specific marker of peroxynitrite （ ONOO - ）. The morphological changes of aortic endothelium were observed under optical microscope and scanning electron microscope （SEM）. The levels of endothelin-1 （ ET-1 ） and thromboxane B2 （TXB2 ） in serum, superoxide dismutase （SOD） and malondialdehyde （ MDA ） in serum were detected. Results： Compared with control group, the iNOS mRNA expression and its protein content, and the NT content all increased significantly in model group,in accordance with the pathological changes of aortic endothelium. NYPBR improved the pathological alterations obviously. Conclusion： NYPBR can decrease iNOS mRNA expression and its protein content, reduce the over-formation of ONOO - and protect the diabetic rats from injury aortic endothelium.