Effect of Danhong injection on expression of nephrin and podocin in podocyte of renal tissue in rats with membranous nephropathy

Abstract

Objective To observe the effect of Danhong injection on the expression of nephrin podocin protein and gene in the kidney of membranous nephropathy rats. Methods A total of 34 male clean level SD rats were divided at random into the normal control group (8 rats), model group (26 rates). The model group were injected the C-BSA into caudal vein to establish the model of the membranous nephropathy. After four week of succession of the models, two model rats were taken to confirm the success of the model. Another 24 models divided at random into model group, Losartan potassium treatment group and Danhong injection treatment group, 8 in each group. In the losartan group, the losartan 40 mg/kg was administered, and the Danhong group was injected with 4 ml/kg of Danhong injection in the tail vein for 4 weeks. The rat urine was collected after the last administration, and 24 h urine protein was determined by spectrophotometer. The serum total protein, albumin, TC and TG levels were determined by automatic biochemical analyzer. The renal pathological changes were observed by HE staining. The expressions of nephrin, podocin and WT1 labeled podocytes were detected by immunohistochemical staining. The expressions of nephrin and podocin in renal tissue were detected by real-time quantitative PCR and Western blot, and the results were correlated. Results Compared with the model group, the content of serum albumin increased in the Danhong group (P<0.01), 24 h urine protein quantitation, TC, TG significantly decreased (P<0.01). The expression of nephrin protein (1.12 ± 0.03 vs. 0.57 ± 0.09), podocin protein (0.99 ± 0.04 vs. 0.53 ± 0.08), nephrin mRNA (0.49 ± 0.08 vs. 0.14 ± 0.02), podocin mRNA (0.50 ± 0.11 vs. 1.18 ± 0.05) in Danhong group significantly increased (P<0.01), and the number of podocytes labeled with WT1 (14.88% ± 1.73% vs. 9.63% ± 1.69%) significantly increased (P<0.05). Correlation analysis showed that 24 h urine protein quantitation and expression of nephrin protein (r=-0.908), podocin protein (r=-0.916), nephrin mRNA (r=-0.930), podocin mRNA (r=-0.923). There was a negative correlation between serum albumin (r=-0.934) and podocyte count (r=-0.778) (P<0.01), and positive correlation with TC (r=0.814) and TG (r=0.870) (P<0.01). Conclusions The Danhong injection may reduce the urine protein of membranous nephropathy rats by up-regulating the expression of nephrin, podocin mRNA and protein in renal cells of membranous nephropathy. Key words: Glomerulonephritis, membranous; Podocytes; Proteinuria; Dan Hong Zhu She Ye; Rats