| Objective: This study established membranous nephropathy rat model by cationic bovine serum albumin(C-BSA),and the modified Shengjiang power and benazepril were applied to the intervention treatment.The serum biochemical index of rats such as blood urea nitrogen,serum creatinine,albumin,total cholesterol and the 24-hour urinary protein quantity were detected.Through observing the renal pathological changes in rats by light microscope,electron microscope,immunofluorescence,detecting the expression of CXCL16 protein and FcRn protein in kidney by immunohistochemical staining and CXCL16 mRNA and FcRn mRNA by real-time PCR,exploring the renal protective effect and the influence of the expression of CXCL16 and FcRn on membranous nephropathy rats of the modified Shengjiang power,theoretical foundation was provided for clinical treatment of membranous nephropathy.Methods: Healthy SD rats 60 of clean level,by feeding a week with standard diet,were randomly divided into the normal group(NG)of 10,the marked group of 50.Making sure 24-hour urinary protein of all the rats were below 5mg.After preimmunization for one week,the tail vein of the marked group was injected with C-BSA three times a week for a total of four weeks.The 24-hour urinary protein was tested.After confirming the success of modeling,the rats of the marked group were divided into five groups: model group(MG),Benazepril treatment group(BG),the modified Shengjiang power treatment group of low dosage(SLG),the modified Shengjiang power treatment group of middle dosage(SMG)and the modified Shengjiang power treatment group of high dosage(SHG).Every group including 10 rats.The rats of BG were given benazepril hydrochloride(10mg/kg/d)grinding dissolved in3 ml saline by intragastric administration.The rats of SLG,SMG and SHG were treated with 3ml(25mg/kg/d,50mg/kg/d,100mg/kg/d),respectively,otherwise the rats of NG and MG were given the equal dose of normal saline.Four weeks later,the rats of every group whose 24-hour urinary protein had been tested already,were cut the abdominal wall and taken blood from the femoral artery,and then the serum biochemical index such as urea nitrogen(BUN),creatinine(Scr),serum total protein(TP),albumin(ALB),total cholesterol(TC),triglyceride(TG)were determined.The fresh kidney tissues were kept.The pathological changes of the kidney tissues were observed by light microscopy,electron microscopy and immunofluorescence.CXCL16 protein and FcRn protein were detected by immunohistochemistry.Real-time PCR was used to detect the expression of CXCL16 mRNA and FcRn mRNA in the kidney.Result:1 General conditionThe rats of NG showed high spirits,quick response and glossy hairs.After molding,the rats of the marked group appeared coarse hairs,low spirits,slow to respond,less ingestion,even abdomen puffy.After medication,the rats in each treatment group were better than before,and no significant improvement in the model group.2 Quantitative comparison of 24 hours urinary protein of rats in each groupAfter molding,24-hour urinary total protein in the marked group was significantly higher than that in the NG,and the difference was statistically significant(P<0.05).After gavage for 4 weeks,the 24h-UTP of each treatment group was significantly lower than that of the model group,and the difference was statistically significant(P<0.05).Compared with SLG,the 24h-UTP level in the MG,SMG,SHG was significantly lower,the difference was statistically significant(P<0.05).But there was no significant difference between the two of the BG,the SMG and the SHG(P>0.05).3 Comparison of serum total protein,serum albumin,total cholesteroland triglyceride in ratsAfter molding,compared with the normal group,the TP and ALB of each marked group were significantly reduced,the TC and TG were significantly increased.The difference was statistically significant(P<0.05).After gavage for 4 weeks,compared with the model group,the treatment group TP and ALB were significantly increased,TC and TG were significantly reduced,the difference was statistically significant(P<0.05);Compared with SLG,the TC and TG in the MG,SMG,SHG were significantly lower,TP and ALB in the MG,SMG,SHG were increased significantly,and the difference was statistically significant(P<0.05);Otherwise there was no statistically significant difference of the levels of TP,ALB,TC and TG among the BG,the SMG and the SHG(P>0.05).4 Comparison of blood urea nitrogen and serum creatinine in every group of ratsTaking the BUN and Scr into account,there were no clear changes among the six groups,as well as there was no significant difference among them(P>0.05).5 The renal pathological changes in every groupNG have no obviously abnormal changes.Under light microscope,MG showed that the basement membrane of glomerular capillary loops had thickened evidently.Under electron microscope,the glomerular basement membrane was irregularly thickened,and a large number of electron dense deposits were deposited.Spike formed.By immunofluorescence appeared that lgG and C3 in granular form deposited diffusively along the capillary loops and mesangial area.Compared with the model group,the rats,pathological changes were slight in the treatment group.Among them,the pathologicalchanges in the modified Shengjiang power of low dosage were between the model group and other treatment groups,and there was no significant difference between other treatment groups.6 The expression of CXCL16 and FcRn in kidney by immunohistochemistryCXCL16 in the normal group and the marked group had different degrees of expression.Compared with the normal group,the expression level of CXCL16 was increased(P<0.05)in each marked group.Compared with the model group,the expression level of CXCL16 in each treatment group was decreased(P<0.05).Compared with SLG,the CXCL16 level in the MG,SMG,SHG was significantly lower,the difference was statistically significant(P<0.05).The expression levels of CXCL16 in the BG,SMG and SHG had no obvious changes,and there was no statistically significant difference between each other(P>0.05).CXCL16 in the normal group and the marked group had different degrees of expression.Compared with the normal group,the expression level of FcRn was increased(P<0.05)in each marked group.Compared with the model group,the expression level of FcRn in each treatment group was decreased(P<0.05).Compared with SLG,the FcRn level in the MG,SMG,SHG was significantly lower,the difference was statistically significant(P<0.05).The expression levels of FcRn in the BG,SMG and SHG had no obvious changes,and there was no statistically significant difference between each other(P>0.05).7 The expression of CXCL16 and FcRn in kidney by real-time PCRCompared with the normal group,the expression levels of CXCL16 mRNA and FcRn mRNA were up-regulated in all marked groups,and the difference was statistically significant(P<0.05).The expression levels of CXCL16 mRNA and FcRn mRNA in all the treatment groups were significantly lower than those in the model group(P<0.05).Compared with SLG,the CXCL16 mRNA and FcRn mRNA in the MG,SMG,SHG were significantly lower,the difference was statistically significant(P<0.05).The expression levels of CXCL16 mRNA and FcRn mRNA in the BG,SMG and SHG had no obvious changes,and there was no statistically significant difference between each other(P>0.05).Conclusions:1 The modified Shengjiang power can increase membranous nephropathyrats,serum total protein and albumin markedly,reduce the membranous nephropathy rats,24-hour urinary protein,total cholesterol and triglyceride significantly.2 The modified powder may have an effect of inhibiting glomerular basement membrane thickening and can immune complex deposition in rats with membranous nephropathy,which can reduce the pathological damage of kidney.3 The modified Shengjiang power can lower the expression of CXCL16 protein and mRNA in nephridial tissue of membranous nephropathy rats,and also lower the expression ofFcRn protein and mRNA.The effect may have something to do with reducing the podocyte injury and delaying the evolve of membranous nephropathy. |
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