养阴活血方药对糖尿病大鼠肾皮质氧化应激损伤的保护作用

Protection of Nourishing Yin and Promoting Blood Circulation Recipe on the Oxidative Stress Injuries in Renal Cortex of Diabetic Rats

目的:通过观察养阴活血方药对糖尿病(DM)大鼠肾皮质一氧化氮合酶(NOS)及过氧亚硝基(ONOO-)水平的影响,探讨养阴活血方药对糖尿病大鼠肾皮质氧化应激损伤的保护作用。方法:将16只SD大鼠采用链脲佐菌素(STZ)复制糖尿病大鼠模型造模,造模成功后,随机分为糖尿病对照组(DM组,n=8)和养阴活血方药组(NYPBR组,n=8),另设8只健康SD大鼠为正常对照组(NC组)。NYPBR组给予养阴活血方药干预。至10周末,检测各组大鼠肾皮质丙二醛(MDA)含量及超氧化物岐化酶(SOD)、总NOS(TNOS)、诱导型NOS(i NOS)和固有型NOS(c NOS)的活性,免疫组化检测肾皮质ONOO-特异性标志物硝基酪氨酸(NT)的生成,检测各组大鼠血糖、体质量、肾质量、24小时尿蛋白及血肌酐。结果:与NC组相比,DM组大鼠肾皮质MDA[(1.78±0.16)nmol/mgprot]水平升高(P<0.05),SOD[(68.96±3.82)U/mgprot]活性下降,TNOS(3.44±0.30)U/mgprot与i NOS(2.54±0.21)U/mgprot活性显著升高,NT[(42.66±3.14)]生成明显增加,肾质量/体质量、24小时尿蛋白、血肌酐水平显著升高。上述指标2组比较差异均有统计学意义(P<0.05)。干预后,BYPBR组肾质量/体质量、24小时尿蛋白、血肌酐较DM组均有明显改善,MDA[(1.56±0.08)nmol/mgprot]下降,SOD[(76.44±4.90)U/mgprot]升高,iNOS[(2.54±0.21)U/mgprot]活性和NT(26.19±1.58)生成均显著降低,上述指标组内比较差异均有统计学意义(P<0.05)。c NOS 3组比较差异无统计学意义(P>0.05)。结论:NYPBR可显著减轻糖尿病大鼠肾皮质氧化应激损伤,此作用可能与提高SOD活性,降低i NOS活性及ONOO-的生成有关。

Objective： To investigate the protection of nourishing Yin and promoting blood circulation recipe（NYPBR） on the oxidative stress injuries in renal cortex of diabetic rats through observing the effect of nourishing Yin and promoting blood circulation recipe on the level of nitric oxide synthase（NOS） and peroxynitrite（ONOO-）in renal cortex of diabetic rats. Methods：Sixteen SD rat models were replicated by streptozotocin（STZ） and divided into diabetes control group（DM group, n=8）, nourishing Yin and promoting blood circulation recipe group（NYPBR group,n=8） and normal control group（NC group） with eight healthy rats at random. The NYPBR group was treated with NYPBR. By the end of the tenth week, the content of renal cortices malondiadehyde（MDA）, the activity of superoxide dismutase（SOD）, total NOS（TNOS）, inducible NOS（i NOS） and constitutive NOS（c NOS） were measured, the generation of nitrotyrosine（NT） as the specific marker of renal cortex ONOO- was detected by immunohistochemical method, blood sugar, body weight, kidney weight, 24-hour urine protein and serum creatinine（SCr） were determined in three groups. Results： Compared with NC group, renal cortex MDA[（1.78±0.16）nmol/mgprot] of DM group elevated（P〈0.05）, the activity of SOD [（68.96±3.82）U/mgprot] decreased（P〈0.05）, the activities of TNOS（3.44±0.30）U/mgprot and i NOS（2.54 ±0.21）U/mgprot elevated significantly（all P〈0.05）, the generation of NT [（42.66 ±3.14）]increased greatly（P〈0.05）, the level of renal mass/body mass, 24-hour urine protein and SCr rose greatly（P〈0.05）.After the intervention, renal mass/ body mass, 24-hour urine protein and SCr improved significantly（ P〈0.05）,MDA[（1.56±0.08）nmol/mgprot] lowered（P〈0.05）,SOD[（76.44±4.90）U/mgprot] elevated（P〈0.05）,the activity of i NOS [（2.54±0.21）U/mgprot] and the generation of NT（26.19 ±1.58） descended greatly（all P〈0.01）.The differences between c NOS（0.70±0.12）U/mgprot and c NOS（0.74±0.13）U/mgprot）of DM group had no statistical significance（P〉0.05）. Conclusion：NYPBR could significantly relieve renal cortex injuries of diabetes rat, which might be related to improving SOD activity, and decreasing i NOS activity and the generation of ONOO-.