补肾活血祛风方对阿霉素肾病大鼠肾病蛋白分子表达的干预研究

Effect of Bushen Huoxue Qufeng Prescription on Nephrin Molecule Expression in Rats with Adriamycin Nephropathy

目的:探讨补肾活血祛风方对阿霉素肾病大鼠足细胞裂隙隔膜分子肾病蛋白(Nephrin)表达及病理组织结构的影响。方法:两次尾静脉注射盐酸多柔比星(阿霉素),制备肾病综合征(Nephrotic Syndrome,NS)阿霉素大鼠模型。造模成功的大鼠再随机分为4组:模型组、激素组、补肾活血祛风方组和激素+补肾活血祛风方组。另设空白组。在造模成功后各干预组灌胃给药,模型组给予等体积蒸馏水灌胃,空白对照组不予干预。第28天测各组大鼠24 h尿蛋白定量、生化指标、免疫组化法检测肾组织Nephrin蛋白水平、PCR检测Nephrin mRNA表达水平、光镜和电镜观察肾组织病理结构变化。结果:与空白组比较,模型组大鼠24 h尿蛋白升高、TC、TG升高(P<0.05);Alb、Nephrin蛋白及其mRNA表达降低(P<0.05);病理可见肾小球肿大、肾小管上皮细胞变性肿胀、间质纤维组织增生等。与模型组比较,激素+补肾活血祛风方组24 h尿蛋白降低最明显(P<0.05),Nephrin mRNA表达升高最明显(P<0.05);补肾活血袪风方组及激素+补肾活血祛风方组TC、TG降低(P<0.05),激素组TC、TG升高(P<0.05);各干预组大鼠肾脏病理不同程度改善,其中激素+补肾活血祛风方组病理病变最轻。结论:补肾活血祛风方可减轻阿霉素肾病大鼠的肾脏病理损害,升高Nephrin表达,降低24 h尿蛋白定量,改善血脂代谢异常,且与激素联合使用效果最优。

Objective:To investigate the effect of Bushen Huoxue Qufeng prescription on the expression of nephrin and the structure of pathological tissue in rats with adriamycin-induced nephropathy.Methods:Doxorubicin hydrochloride(Adriamycin)was injected twice through the tail vein to prepare a rat model of nephrotic syndrome(NS)with Adriamycin.The successfully modeled rats were randomly divided into 4 groups:model group,hormone group,Bushen Huoxue Qufeng prescription group and hormone+Bushen Huoxue Qufeng prescription group.Set another blank group.After the successful model building,each intervention group was given intragastric administration,the model group was given distilled water intragastric administration,and the blank control group received no intervention.On the 28th day,the 24 h urine protein quantification,biochemical indicators,immunohistochemical method to detect nephrin protein content in renal tissues,PCR to detect nephrin mRNA expression content,and light microscope and electron microscope to observe the pathological structure of renal tissue were measured.Results:Compared with the blank group,the 24 h urine protein and TC and TG of the model group increased(P<0.05);the expression of Alb,nephrin protein and mRNA decreased(P<0.05);the pathology showed glomerular enlargement and renal tubules Epithelial cell degeneration and swelling,interstitial fibrous tissue hyperplasia,etc.Compared with the model group,the hormone+Bushen Huoxue Qufeng Prescription group had the most significant reduction in 24 h urine protein(P<0.05),and the increase in nephrin mRNA expression was the most significant(P<0.05);TC and TG decreased in the wind prescription group(P<0.05),and TC and TG increased in the hormone group(P<0.05);the renal pathology of rats in each intervention group improved to varying degrees,and the hormone+Bushen Huoxue Qufeng prescription group had the least pathological changes.Conclusion:Bushen Huoxue Qufeng prescription can alleviate the pathological damage of the kidney in rats with adriamycin nephropathy,increase the expression of nephrin,reduce the 24 h urine protein quantification,improve the abnormal blood lipid metabolism,and the combination of hormones has the best effect.