| Obiective:to research5/6nephrectomy chronic renal failure rats renal epithelial cells EMT to renal fibrosis mechanism, and explore the efficacy mechanism of Liuwei Dihuang decoction against renal fibrosis.Methods:Healthy male Sprapue-Demey rats were randomly divided into normal group, sham operation group and operation group. Experimental rats after adaptive feeding for a week.Copy the5/6nephrectomy rat model of chronic renal failure. All operations were carried out under sterile conditions, the surgical group completed two operations, the first time the left kidney, the lower pole resection in1/3, a week after the completion of the second right kidney removal surgery, the sham operation group, only to open the abdominal cavity,The normal group without any treatment.One week after the removal of the right kidney, the survival of operated rats were randomly divided into model group, Liuwei Dihuang decoction group.The Liuwei group every day with Liuwei Dihuang decoction gavage, model group, sham group and normal group to distilled water. All animals were sacrificed after8weeks intragastrically. Prior to modeling, modeling after1-9weeks, weighing every two weeks. Before the animals were killed to collect blood to test BUN and CR levels, HE staining, Masson staining observed morphological changes of the left remnant kidney. Immunohistochemistry, Western blot, RT-PCR methods were used to detect E-cadherin, α-SMA, FSP-1, TGF-β1, BMP-7expression differences of the rat left remnant kidney at the protein and mRNA levels,in cell morphology and molecular biology level of differential expression.Results:①Compared with normal group:model group, body weight slowly increased with the feeding times are extended, but the growth rate was significantly later, the difference was statistically significant (P<0.01);24h urine volume and urinary protein were significantly higher (P<0.01);Renal function, BUN, Cr were significantly increased (P<0.01); Retained renal naked eye to change as the texture is hard and tough, capsule difficult to peel, cut renal cortical thinning. Microscopic change capsule thickening fibers, most of the glomerular atrophy, fibrosis, and renal capsule narrow, a small number of glomerular hypertrophy, the majority of renal tubular epithelial cell atrophy, off, part of the lumen irregular expansion, there are a lot of protein in tube; renal interstitial fibrous connective tissue hyperplasia, apparent renal fibrosis; model group in the protein and mRNA levels, the epithelial cell marker E-cadherin expression level was significantly down-regulated and the muscle fiber cell marker α-SMA and FSP-1expression were significantly up-regulated, the difference between the above group were significant statistically significancet (P<0.01), EMT phenomenon appears, and TGF-β1expression levels increased, BMP-7expression levels lowered, the difference was statistically significance (P<0.01);②Compared with model group after8weeks Liuwei Dihuang decoction treatment:Liuwei group of rats body weight increased significantly,24h urine volume and urinary protein were lower,BUN was lower, the difference between the above groups were statistically significant (P<0.01), renal interstitial fibrosis to reduce, in the protein and mRNA levels, E-cadherin expression was up, α-SMA, FSP-1expression were down, EMT phenomena reduced, TGF-β1expression down-regulated, the expression of BMP-7level raised,above differences between the groups was statistically significant (P <0.01). All of the above experimental results, the sham operation group compared with normal group. was no significant difference (P>0.05).Conelusion:①successfully copied5/6nephrectomy induced rat model of chronic renal failure;②confirmed EMT phenomenon in the rat remnant kidney of5/6nephrectomy-induced chronic renal failure model;③rat remnant kidney EMT in chronic renal failure model caused by5/6nephrectomy may be associated with TGF-β1upregulation and BMP-7downregulation in expression levels; the④Liuwei Dihuang decoction reduce the remnant renal pathological changes, and renal interstitial fibrosis;⑤The Liuwei Dihuang decoction inhibit the EMT;⑥The efficacy mechanism of Liuwei Dihuang decoction inhibiting the EMT and antagonising renal interstitial fibrosis may be by down-regulating TGF-β1and up-regulating BMP-7expression levels. |
PDF Full Text Request