Haikunshenxi Rats With Adriamycin Nephrosis Of Endothelin-1 Regulation Mechanism

Recently, researchers all of the world had payed close attention to the role of endothelin(ET) in the Chronic renal failure(CRF).Some of them proposed that the ET level reflected the severity of CRF, because increased levels of ET have been found in CRF nephridial tissue,blood and urino routine examination. Glomerular mesangial cells(GMCs), glomerular epithelial cell (GEC) and glomerular vascular endothelial cell (GVEC) generated more ET-1 when kidney suffered kinds of inflammation, and tubuli epithelial cell excreted more ET-1 to promote nephric hypofunction and fibrosis progressively. Nephric fibrosis was the common pathway in which various kinds kidney disease developed CRF, including glomerular sclerosis and renal interstitium fibrosis. Therefore, it was significant to research the role of ET in the mechanism of nephric fibrosis."Haikunshenxi"was the nationalâ…¡grade new Chinese medicine to treat CRF and early-stage uremia. It was composed by Fucoidan(FPS). The researches showed that FPS had many functions of biology, such as stepping the blood pressure, blood fat and blood sugar down, enhancing immunity and so on. To elucidate the mechanism of action of Haikunshenxi preventing and curing CRF, we researched it from levels of tissue, cell, protein and gene by animal experiment and vitro cell experimentObjectiveIn order to elucidate the mechanism of Haikunshenxi preventing and curing CRF, we observed effectivity of Haikunshenxi by established Adriamycin nephropathy rats model. We also researched the effection of Haikunshenxi to ET-1 from levels of organ, tissue, cell, protein and gene.Method1.SP male wistar rats were randomizated according to weights. Then the operation groups performed right kidney exsection and injected intraperitoneally to induce CRF model. Each treatment group was treated with homologous medicine. Rats'weight was measured weekly, and 24h Urine protein was measured every two weeks. The kidney was taken at the 10th week to be observed the changes of the pathomorphology, and be estimated the pathology changes by glomerular sclerosis index number and tubuli renales-mesenchymal injure score , at the same time the blood serum was separated to examination blood biochemical indicator.2.The ET-1 protein was detected in rats nephridial tissue by immunohisto- chemical method; ET-1mRNA level was detected by hybridization in situ method.3.With LPS stimulating GMCs, the proliferation was detected by MTT method at 12h, 24h, 48h and 60h, and the expression of ET-1mRNA was detected by RT-PCR.4.With LPS stimulating GMCs, the ET-1 proteic content was detected by immunohistochemical method and western blot method.Results1.After the Haikunshenxi treatment, the weight of rats rised conspicuously, 24h Urine protein reduced, BUN , Scr , Cho, TG dropped obviously. The changes of pathomorphology of rats kidney was ameliorated obviously.2.Haikunshenxi declined the expression of the ET-1 protein and the ET-1mRNA in kytoplasm of nephridial inherent cells of adriamyc nephronia rats.3.Haikunshenxi suppressed the multiplication of GMCs stimulated by LPS , the large dose suppression effect was the best. The RT-PCR experimental result indicated that Haikunshenxi may remarkably suppress the expression of ET-1mRNA in GMCs, and showed the dosage dependence relations.4.Haikunshenxi suppressed GMCs to secrete the ET-1 protein obviously, and showed the dosage dependence relations.Conclusion1.Haikunshenxi improved the kidney function of adriamyc nephronia rats, reduced the urine protein, reduced the pathology damage of kidney, and showed the dosage dependence relations.2.The mechanism of Haikunshenxi prevented and cured kidney fibrosis. In cellular level, Haikunshenxi suppressed the multiplication of GMCs, which was possibly one of the mechanisms that Haikunshenxi prevented and cured glomerular sclerosis. In the protein level, Haikunshenxi declined the ET-1 protein expression in kidney organization and GMCs, then decreased the risk of the kidney fibrosis. In the gene level, Haikunshenxi suppressed the expression of ET-1mRNA through reverse transcription, and that suppressed the multiplication of GMCs, and reduced ECM, thus delayed CRF.