复方积雪草对糖尿病肾病大鼠肾脏保护作用及对血红素加氧酶-1、NADPH氧化酶4蛋白表达的影响

Effect of Compound Centella on Renal Protection and Protein Expression of HO-1,NOX4 in Diabetic Nephropathy Rats

目的:观察复方积雪草对STZ联合高脂高糖饲料诱导的糖尿病肾病大鼠肾脏保护作用及对肾组织中血红素加氧酶-1(HO-1)、NADPH氧化酶4(NOX4)蛋白表达的影响。方法:20只SD大鼠随机分为复方积雪草组、氯沙坦钾组、模型组和正常对照组,每组5只大鼠。复方积雪草组、氯沙坦钾组、模型组大鼠从实验第1天起至第112天,全程用高脂高糖饲料喂养,并在实验第29天给予单次腹腔注射链脲霉素(STZ)35 mg/kg。实验第112天处死所有大鼠。对大鼠空腹血糖、24 h尿蛋白定量、血肌酐进行检测。予HE及PAS染色观察肾脏病理改变,透射电镜观察肾脏超微结构。氧化试剂盒检测超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量。Westernblot检测肾组织中HO-1、NOX4蛋白含量。免疫组化观察肾组织中HO-1、NOX4蛋白表达部位及强度。结果:在模型组中,24 h尿蛋白定量增多,光镜可见小球体积增大,系膜及基质增生,肾小管空泡变性,透射电镜可见足细胞融合脱落,肾组织中SOD活性下降、MDA含量增高、HO-1蛋白表达减少、NOX4蛋白表达增多。经过复方积雪草干预,24 h尿蛋白减少,肾病理改善,肾组织中SOD活性升高,MDA含量减少,HO-1蛋白表达上调,NOX4蛋白表达下调。氯沙坦钾组中可观察到类似结果。结论:复方积雪草可能通过调节氧化应激关键蛋白HO-1及NOX4的表达从而保护糖尿病肾病。

Objective:To observe the effect of compound centella on the renal protection of diabetic nephropathy rats induced by STZ combined with high-fat and high-sugar diet and the protein expression of HO-1,NOX4 in renal tissue.Methods:20 SD rats were randomly divided into the compound Centella group,losartan group,model group and normal group,with 5 rats in each group.Rats in the compound Centella group,losartan group and model group were fed with high-fat and high-sugar diet from day 1 to day 112 and were given a single intraperitoneal injection of STZ 35 mg/kg on day 29.All rats were killed on day 112 of the experiment.Fasting blood glucose,24-hour urinary protein quantification and serum creatinine were measured.HE and PAS staining were used to observe the pathological changes of the kidney and the ultrastructure of the kidney was observed by transmission electron microscopy.The SOD activity and the MDA content were detected by oxidation kit.Westernblot was used to detect the protein expression of HO-1 and NOX4.Immunohistochemistry was used to observe the expression sites and contents of HO-1 and NOX4 proteins in renal tissues.Results:In the model group,24-hour urinary protein increased.The mesangial cells and stroma were proliferated.Moreover,renal tubule vacuolization was also observed in the model group.The SOD activity decreased but the MDA content increased in renal tissues.The HO-1 protein expression decreased while the NOX4 protein expression increased.After the treatment of compound Centella,24-hour urinary protein decreased and renal pathology improved.The SOD activity increased and the MDA content decreased.The HO-1 protein expression up-regulated while the NOX4 protein expression down-regulated.Similar results were observed in the losartan group.Conclusion:Compound Centella may protect diabetic nephropathy by regulating the expression of key oxidative stress proteins HO-1 and NOX4.