黄芪对IgA肾病大鼠蛋白尿及肾组织表达nephrin、podocin的影响

Effect of Huangqi on Proteinuria and Nephrin,Podocin in Rats with IgA Nephropathy

目的观察黄芪对IgA肾病(IgAN)大鼠蛋白尿及肾组织nephrin、podocin的影响,探讨黄芪对IgAN蛋白尿的治疗作用及其机制。方法 24只6周龄SD大鼠分为对照组、模型组及黄芪组。模型组和黄芪组采用牛血清清蛋白+脂多糖+四氯化碳的方法进行IgAN造模,黄芪组予黄芪颗粒治疗,对照组予等量蒸馏水灌胃,9 g.L-1盐水皮下注射及尾静脉注射。考马斯亮蓝法检测其24 h尿蛋白定量,全自动生化分析仪检测其血生化指标。应用直接免疫荧光法检测其肾小球IgA沉积强度,应用间接免疫荧光法检测肾小球nephrin、podocin蛋白的表达及分布,采用实时荧光定量PCR技术检测肾皮质nephrin mRNA和podocin mRNA的表达。结果 1.黄芪组大鼠尿蛋白较模型组显著减少,差异有统计学意义(P<0.01)。2.黄芪组大鼠肾组织病变较模型组减轻。3.黄芪组大鼠肾小球nephrin表达量与模型组比较差异无统计学意义(P>0.05),但仍显著高于对照组(P<0.01);而podocin表达量较模型组显著降低(P<0.05);黄芪组大鼠肾皮质nephrin mRNA及podocin mRNA的表达均较模型组及对照组显著升高(Pa<0.01)。4.黄芪组大鼠肾小球nephrin、podocin的不连续斑片状、团块状异常分布较模型组改善。结论 IgAN大鼠肾组织nephrin、podocin出现表达变化及分布异常,黄芪能减少IgAN大鼠尿蛋白,其作用可能与调节nephrin、podocin的表达及分布有关。

Objective To observe the effect of Huangqi on proteinuria in rats with IgA nephropathy（IgAN） and dynamic changes of expression and distribution of nephrin and podocin,and to further explore the mechanism of proteinuria of IgAN and try to find the approach to the therapy for IgAN.Methods SD rats with 6 weeks（n=24） were randomly divided into 3 groups：control group（n=8）,model group（n=8） and Huangqi group（n=8）.IgAN model was induced by the improved method of bovine serum albumin＋carbon tetrachloride＋lipopolysaccharide.Huangqi group was given Huangqi grain for treatment,but control group and model group were given distilled water.Quantitative urinary protein of 24 hours was tested by Coomassie brilliant blue assay,and blood biochemistry indexes were tested by automatic biochemistry analyzer.The IgA intensity was examined by direct immunofluorescence.The protein expression and distribution of nephrin and podocin were examined by indirect immunofluorescence.The mRNA expression of nephrin and podocin were examined by real-time polymerase chain reaction.Results 1.Urinary protein in Huangqi group was less than that in model group（P〈0.01）.2.Pathological changes of nephritic tissue in Huangqi group were less serious than those in model group.3.The protein expression of nephrin in Huangqi group had no significant diffe-rence compared with that in model group（P〈0.05）,but higher than that in model group（P〈0.01）.Podocin protein expression in Huangqi group was less than that in model group.In Huangqi group,mRNA of nephrin and podocin expressed markedly higher than those in model group and control group（Pa0.01）.4.Abnormal discontinuous distribution of nephrin and podocin in Huangqi group was slighter than that in model group.Conclusions Changes of expression and abnormality of distribution of nephrin and podocin appeared in rats with IgAN.Huangqi is good for decreasing urinary protein by the way of regulating the expression and distribution of nephrin and podocin.