| Objective:In this experiment, through established IgA nephropathyanimal model, application of Yiqiyangyin Chinese medicine treatment ofanimal model, and to observe the experimental animal24hour urinary proteinquantity, the red blood cell count,renal function (SCr), renal pathologicalchanges, immunohistochemistry and PCR method in order to examin theexpression of ACE, radioimmunoassay method was used to detect the amountof serum AngII,to study the therapeutic effect of Yiqiyangyin inesemedicine treatment on IgA nephropathy and its possible mechanism, providenew theoretical and experimental basis for the clinical application.Method:40SPF male Sprague-Dauley rats with about150g weref-ed adaptively for1week, and then they were randomly divided into nor-malgroup, model group,Yiqiyangyin group and Benazepril group,10ger-bils eachgroup.Except the normal group, all the other rats use the foll-owingmethods:Took orally immunogen BSA (Sigma, USA),configured t-o100g/L,at adose of400mg/kg.qod for8weeks.Subcutaneous injectionofcarbon tet-rachloride0.1ml, castor oil0.3ml for once a week and co-ntinuous9weeks. At the6thweek, injecting LPS(Sigma, USA)0.05mg.Normal groupwere to be fed the same amount of distilled water for8weeks.Once a week forsubcutaneous injection of saline solution0.4ml for nine consecutive weeksand in the sixth week tail intravenous injec-tion amount ofsaline solution.Therats of24h of urine were collected w-ith metabolic cages and whose urineprotein obviously increased enter th-e stage of treatment. The10th week,Yiqiyangyin group and Benazepril group respectively to bay that splithydrochloride,corresponding Chinese medicine lavage, normal group, modelgroup give amount of saline lavage, for8weeks.All rats were put intometabolsm cages to obtain24hour urine to measure24hour urinary protein quantitative and red blood cell countbefore modeling and at3thweek,6thweek,9thweek,13thweek,17thweekafter modeling respectively. At the end ofthe17thweek,all animals werekilled to obtain the blood and the renal tissues,Scr and BUN were ass-ayed with the automaticbiochemistry analyzer.Pathologic changes of glo-merulus by light microscopy. Analysising ACE innephridial tissue by i-mmunohistochemistry and RT-PCR. Analysising AngIIin serum by radio-immunity.Results:1The24hour urinary protein quantity in each groupThe24hour urinary protein quanty of the rats in normal group ha-d no obvious differences at the times of the end of the3th,6th,9th,13th,17thweek (P>0.05).Rats of model group and treatment groups after building6weeken-d began to appear proteinuria, and urine protein with progressive growt-h,on the9th weekend reached the highest. Compared with the controlgroup, the difference was statistically significant (P<0.05).<0.01).After treatment, the rats of treatment groups of urine protein beganto decline after4weeks. Compared with model group,24hours urinaryprotein quanty was reduced (P<0.05).There was no significant differencebetween the Yiqiyangyin group and Benazepril group (P>0.05).inapuli g2Urine red blood cell count in each groupRats of normal group in the experiment, all did not appear blood in theurine.Rats of model group and treatment groups after building6weekendbegan to appear hematuria, and the urine red blood cell count with pr-ogressive growth, on the9th weekend reached the highest.Compared w-ith the control group,the difference was statistically significant (P<0.05)After treatment, the rats of Yiqiyangyin group of urine red blood c-ellcount began to decline after4weeks and compared with model grou-p urine red blood cell count was reduced(P<0.05).Compared with modelgroup,the urine red blood cell count of Benazepril was reduced slightly. But there was no significant difference between the two groups.(P>0.05).3Scr and BUN of each groupThe Scr and BUN were no differences in each group (P>0.05).4The pathomorphology changes of kidneyObservations under light microscope:In the normal group: the struct-ure of renal glomerulus was integrated,glomerular mesenterium and basematerial were normal. In the model group: thickening glomerular mesan-gial cell proliferation thickening, mesangial matrix increased, mesangialarea broadening.The Yiqiyangyin group and Benazepril group also show-ed different degree of pathological changes, but the pathological degreewas obviously lighter than the changes of the model group.Observations under immunofluorescence:In the normal group: the gl-omerular mesenterium had no immune complex deposition. In the modelgroup: the immune complex deposited granularly along the glomerularmesenterium. The Yiqiyangyin group and Benazepril group also showeddifferent degree of immune complex deposition, but the degree was ligh-ter than the changes of the model group.5RT-PCR and immunohistochemical results of the expression of ACE inkidneyRT-PCR method:Compared with the normal group, model group andtreatment groups, ACE expression was significantly increased (P<0.05);compared with the model group,the expression of ACE in the treatmentgroups was decreased (P<0.05).Immunohistochemical results:Compared with the normal group, modelgroup and treatment groups, ACE expression was significantlyincreased(P<0.05); compared with the model group, the expression of ACE in thetreatment groups was decreased(P<0.05).6AngII amount determined by radioimmunoassayCompared with the normal group,model group and treatment groups,ACE expression was significantly increased (P<0.05); compared withthemodel group, the expression of ACE in the treatment groups wasdecree-sed (P<0.05).Conclusions:1Yiqiyangyin prescription could decreas the proteinuria and urine redblood cell count of IgAN rats.2Yiqiyangyin prescription could reduce the pathological damages of kid-ney tissues.3Yiqiyangyin prescription can inhibit the expression of ACE in kidney tissueand to reduce the amount of AngII in blood, this hints it may inhibit theactivation of RAS system protect the kidney disease progress. |
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