| Objective: To observe the effects of different doses of Yishen-Tongluo Prescription on the general conditions,biochemical indexes and renal pathology of rats with membranous nephropathy through animal experiments,so as to provide a theoretical basis for clinical treatment of membranous nephropathy.Methods:1.Sixty healthy male SD rats were selected,and after one week of adaptive feeding,24-hour urinary total protein(24hUTP)was negative(<5mg)and randomly divided into normal group(NG,n=10)and modeling group(n=50).2.The model group was established by the tail vein injection of cationic bovine serum albumin(C-BSA),after the end,measured 24 hUTP,modeling success.3.Model rats were randomly divided into model group(MG),western medicine group(BG),traditional Chinese medicine low dose group(ZLG),traditional Chinese medicine middle dose group(ZMG),traditional Chinese medicine high dose group(ZHG),each group 10 only.4.The traditional Chinese medicine group was treated with Yishen Tongluo decoction(6.61g/kg/d?13.22g/kg/d?26.44g/kg/d)orally,and the western medicine group was given benazepril hydrochloride(10mg/kg/d).The model group and the normal group were given equal volumes of distilled water for 4 weeks.5.Observe the general condition of each group of rats.After 4 weeks of treatment,measure 24 hUTP in each group,take blood from abdominal aorta,measure blood biochemical indexes,and take kidney tissue.Observepathological changes of kidney in rats by light microscope,immunofluorescence and electron microscope.The expression of ezrin and synaptopodin mRNA in the kidney was measured by real-time PCR and histological analysis of ezrin and synaptopodin protein.Results:1.General conditionCompared with NG,MG rats had poor luster,apathy,unresponsiveness,and decreased feeding,and some had abdominal edema.Compared with the model group,the condition of the rats in each treatment group improved.2.24h-UTP comparison of rats in each groupCompared with NG,24h-UTP in model group increased significantly(P<0.05).The treatment group was significantly lower than MG(P<0.05);compared with BG,ZLG was higher(P<0.05),ZMG,ZHG had no significant difference(P>0.05);compared with ZLG,ZMG,ZHG decreased significantly(P<0.05).3.Comparison of blood biochemical indexes in each groupCompared with NG,the TP and ALB were significantly lower in the modeling group,and TC and TG were significantly increased(P<0.05).Compared with MG,TP and ALB were significantly higher in each treatment group,and TC and TG were significantly lower(P<0.05).Compared with BG,TP and ALB of ZLG decreased,and TC and TG increased(P<0.05).ZMG,ZHG had no significant difference(P>0.05);compared with ZLG,ZMG,ZHG decreased significantly(P<0.05).There was no significant difference in BUN and Scr between rats in each group(P>0.05).4.Kidney pathological changes in each groupThere was no significant change in the glomerular structure of NG rats.MG rats under light microscope epithelial subparietal deposition of eosinophils,diffuse thickening of the basement membrane,nail formation;the treatment group pathological changes are minor.Immunofluorescence: NG glomeruli showed no fluorescence;MG saw lgG,C3 granular deposition along the diffuse capillary deposition,fluorescence intensity was obvious;the fluorescence intensity of the treatment group was weak.Electron microscopy showed that the NG foot process was clear and well-arranged;the basement membrane of MG was diffused and thickened,a large number of electron dense materials were deposited under the epithelium,and the foot processes were extensively fused or disappeared,and the nail processes formed.The treatment group was slightly more pathological than the model group.5.ImmunohistochmistryNG: ezrin and synaptopodin are mainly expressed in the glomerulus and are dotted along the capillary loop.Ezrin is expressed in small amounts in the renal tubules.Compared with NG,the expression of MG was decreased(P<0.05).Compared with MG,the expression of both of them was enhanced(P<0.05);compared with BG,the expression of both of them was weak(P<0.05),ZMG,ZHG difference was not significant(P>0.05);compared with ZLG,ZMG,ZHG expression increased(P<0.05).6.Real-time PCRCompared with NG,the expression of ezrin and synaptopodin mRNA in MG was down-regulated(P<0.05).Compared with MG,the expression of ezrin and synaptopodin was up-regulated in all treatment groups(P<0.05);compared with BG,the expression of ezrin and synaptopodin was lower in ZLG;the expression levels of ZMG and ZHG were not significantly different(P<0.05).ZMG and ZHG were up-regulated compared with ZLG(P<0.05).Conclusions:1.Yishen Tongluo decoction can significantly increase the levels of TP and ALB in rats with membranous nephropathy,and significantly reduce24 hUTP,TC and TG levels in membranous nephropathy rats.2.Yishen Tongluo decoction may inhibit the deposition of glomerular basement membrane immune complexes and the thickening of basement membrane in rats with membranous nephropathy,and may reduce the renal pathological injury such as foot process fusion.3.Yishen Tongluo decoction may up-regulate the expression of ezrin and synaptopodin proteins,ezrin and synaptopodin mRNA in renal tissue of rats with membranous nephropathy,thus reducing podocyte injury,reducing proteinuria,protecting renal tissue,and delaying the progression of membranous nephropathy. |
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