| Objective: IgA nephropathy(IgAN)is a group of primary glomerular disease,characterized by basing on IgA immune complex depositing in glomerular mesangial region,clinical and pathological manifestations are diverse.At present,there is no ideal treatment for western medicine,but in recent years,the curative effect of traditional Chinese medicine on IgAN has been affirmed.In this experiment,by making the model of Ig AN rats,observe the effects of Shutong Sanjiao Qingre Lishi recipe on kidney TGF-?1 and Serum IL-6?IL-17 expression levels of IgAN rats,and investigate the possible nosogenesis of Shutong Sanjiao Qingre Lishi recipe on IgAN rats to provide experimental theoretical basis for clinical treatment of IgAN.Methods: 40 Sprague-Dauley(SD)rats,weighing about 150~180g,were placed in a laboratory room.The rats were fed adaptively for one week and their urine protein or occult blood was negative.Then they were randomly divided into four groups: Normal group(N group)?Nmodel group(M group)?benazepril group(X group)and traditional Chinese medicine group(Z group),10 rats in each group.The method of making IgAN rat model by Zhang Jing is made by using the combined method of BSA,LPS and CCL4.After the completion of the model,according to the grouping,the corresponding drug intervention was given respectively:Z group was given Shutong Sanjiao Qingre Lishi recipe and X group was given benazepril hydrochloride daily for8 weeks.The urine samples of rats were collected at the 2nd,6th,11 th,15th and the 19 th week respectively to detect 24h-UTP.At the begin of the 19 th week of the experiment,the rats were fasting water for 12 hours and weighed.Then there were anesthetized with chloral hydrate,blood was taken from femoral artery to check the content of Scr?BUN and ALB.Renal tissue was taken and stained with HE ? Masson and PASM,and immunofluorescenceassay was used to detect the deposition of immune complexes IgA in rat kidney.The expression of TGF-?1 in renal tissue was observed by immunohistochemistry and RT-PCR.The content of IL-6 and IL-17 in serum of experimental rats were detected by ELISA.Results:1 The comparison of 24h-UTPBefore modeling,the urine protein was negative in all experimental rats.After starting the modeling,compared with N group,the 24h-UTP in M?X?Z group were significantly higher,and the difference was statistically significant(P<0.05),but there were no significant change in M ? X ? Z group(P>0.05).After treatment with benazepril and Shutong Sanjiao Qingre Lishi recipe,the 24h-UTP in M?X?Z group were higher compared with N group,and the difference was statistically significant(P<0.05).But compared with M group,the 24h-UTP in X?Z group were significantly lower(P<0.05),and there were no significant change in X?Z group(P>0.05).2 The content comparison of Scr?BUN and ALBThere was no significant difference in serum levels of Scr?BUN and ALB(P>0.05).3 Pathological observation of renal tissue3.1 The observation by light microscopeIn N group,the glomerular structure was intact,the glomerular Mesangium and matrix were no obvious abnormal changes.In M group,the renal tissue showed obvious mesangial cell hyperplasia,and mesangial matrix increased.In X ? Z group,they were the same to the M group,but the pathological changes were lighter than those in M group.3.2 Immunofluorescence observationIn N group,there were no immune complex founded in glomerular mesangial region.In group M,massive fluorescent green immune complexes were deposited in the glomerular Mesangial region.The immune complex was deposited in the glomerular Mesangial area of the experimental rats in the X?Z group,but the deposition amount was significantly less than that in the Mgroup.4 The expression of TGF-?1 in Renal tissue by the methods of Immunohistochemical and RT-PCR4.1 Immunohistochemical methodCompared with N group,the expression of TGF-?1 in renal tissue in M?X?Z group was significantly higher(P<0.05).Compared with M group,the expression of TGF-?1 in X?Z group was decreased(P<0.05),but there were no significant change in X?Z group(P>0.05).4.2 RT-PCR methodCompared with N group,the expression of TGF-?1 mRNA in renal tissue in M?X?Z group was significantly higher(P<0.05).Compared with M group,the expression of TGF-?1 mRNA in X?Z group was decreased(P<0.05),but there were no significant change in X?Z group(P>0.05).5 By ELISA method to check the content of IL-6 and IL-17 in rat serumCompared with N group,the serum IL-6 and IL-17 levels in M?X?Z group were significantly higher(P<0.05).Compared with M group,the expression of IL-6 and IL-17 in X?Z group was decreased(P<0.05),but there were no significant change in X?Z group(P>0.05).Conclusions:1 Compared with benazepril hydrochloride,Shutong Sanjiao Qingre Lishi Recipe can significantly reduce the 24h-UTP in IgAN rats.2 Shutong Sanjiao Qingre Lishi Recipe can reduce renal pathological injury in Ig AN and delay the further development of IgAN.3 Its mechanism for retarding IgAN progression may be closely related to reducing the expression of TGF-?1 in the renal tissue and reducing the content of IL-6 and IL-17 in the IgAN serum. |
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