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Effects of Supplementing Qi, Nourishing Yin Method on the Expression of Type Ⅳ Collagen and AT1in Rats of IgA Nephropathy

Author: AnRan
Tutor: GuoDengZhou
School: Hebei Medical University
Course: Traditional Chinese Medicine
Keywords: IgA nephropathy Renin-angiotensin system Angiotensin Ⅱ Angiotensin Ⅱ typeⅠreceptor (AT1) Type Ⅳcollagen Glomerular sclerosis
CLC: R692.3
Type: Master's thesis
Year: 2014
Downloads: 2
Quote: 0
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Objective: Immunoglobulin A (IgA) nephropathy is the mostcommon primary glomerular disease worldwide which seriously threatensthe health of patients. In recent years, studies have found that15%~20%of patients within10years,30%~35%of patients within20years afterthe diagnosis will develop into end-stage kidney failure. IgAN has higherincidence in our country, the clinical manifestations are diverse, whichcharacterized by recurrent hematuria. It can be associated with proteinuria,hypertension,etc. The diagnosis of IgAN is currently depended onpathologic examination combined with clinical manifestations such ashematuria and proteinuria. Its histopathologic characteristics mainlyinclude depositions of polymeric IgA1molecules in glomerularmesangium with a small amount of C3, IgM and IgG depositions andmesangial cells proliferation and mesangial matrix increasing. About itspathogenesis,it is still poorly understood, many scholars considered thatit is the comprehensive result of many factors. In resent years,the role ofcytokines caused for concern. Studies have found that the activation ofthe RAS system in local kidney can stimulate the mesangial cellsproliferation, mesangial matrix broadening and the synthesis ofextracellular matrix which plays an important role in the progressiveprocess of IgAN. At present our therapeutic armamentarium is stilllimited largely to supportive care and immunosuppression in someinstances, so discovering specific and effective therapeutic strategybecomes a research hotspot. So we put forward ‘supplementing qi,nourishing yin’ method to treat IgAN and then obtained good therapeuticeffects after clinical application, but its specific mechanism remained unclear.In this paper, we firstly established the animal model of IgAN. Thenwe carried on intervention treatment according to the corresponding dosewith traditional Chinese medicine of supplementing qi, nourishing yin.Through observing24-hour urinary protein quantitative, urine red bloodcell count, renal function and detecting the level of AngⅡand expressionof AT1, type Ⅳ collagen in renal tissue,we aimed to explore the possiblemechanism and drug targets of this method in the treatment of IgAnephropathy.Methods: Forty male Sprague-Dawley rats in clean level wereselected. All rats were randomly divided into four groups:10in normalcontrol group,10in model group,10in benazepril group,10intraditional Chinese medicine group. The rats in last three groups inaddition to control group were established animal model of IgAnephropathy with bovine serum albumin, lipopolysaccharide, carbontetrachloride and castor oil. The specific method is as follows: First, weprepared the BSA solution of100g/l concentration with distilled water.Then the rats were treated with BSA by gavage at the dosage of400mg/kg every other day for eight weeks; they were injected carbontetrachloride0.1ml and castor oil0.3ml by subcutaneous injection oncea week for nine weeks; at the6th group, the rats were givenlipopolysaccharide by tail vein injection0.05mg one time. The rats incontrol group were gave distilled water by gavage with the same doseevery other day for eight weeks; They were injected physiological saline0.4ml by subcutaneous injection once a week for nine weeks; At the6thgroup, the rats were injected physiological saline0.2ml by tail veininjection one time. Then we gave the rats of treatment group benazeprilhydrochloride and traditional Chinese medicine by gavage respectivelywith the corresponding quantity from the10th week for8weeks. Theother group rats were gave distilled water by gavage at the same doseevery day for eight weeks. In this process, we put the rats in the metabolic cage to obtain urine specimens at the each time point of theexperiment.Then we detected red blood cell count and24hours urinaryprotein quantitative. At the17thweeks, we anesthetized all the rats andthen took the blood and collected kidney specimens quickly. Then wedetected renal function of centrifugal blood with semi-automaticbiochemical analyzer and the expression level of AngⅡ withradioimmunity technique. At the end of the experiment, we observedhistopathological changes of kidney under light microscope as well aselectron microscope, and detected the expression levels of IgA immunecomplex deposition, type Ⅳ collagen and AT1gene by applying withimmunofluorescence method immunohistochemical technique andreverse transcription polymerase chain reaction technique respectively.Results:Compared with normal group,AT1gene expression of the rats inmodel group, benazepril group, traditional Chinese medicine group wereincreased significantly (P <0.01).This index of rats in benazepril group,traditional Chinese medicine group was obviously lower than that inmodel group (P <0.01). But we observed no obvious difference betweenbenazepril group and traditional Chinese medicine group.In normal group, type Ⅳ collagen was mainly expressed in theglomerular and renal tubular basement membrane of the rats. Mesangialarea renal interstitial almost had no expression. The expression of type Ⅳcollagen of rats in model group,benazepril group and traditional Chinesemedicine were significantly increased compared with normal group. Itwas visible in the mesangial area. The expression of type Ⅳcollagen ofrats in benazepril group and traditional Chinese medicine group wassignificantly reduced compared with model group. But there were nosignificant differences between benazepril group and traditional Chinesemedicine group.Conclusion:1Traditional Chinese medicine of supplementing qi, nourishing yin can lower the urine protein and reduce urine red blood cell count in ratsof IgA nephropathy to protect renal function;2This medicine can significantly improve the pathological lesion inrats of IgA nephropathy, such as the foot process fusion of podocytes,mesangial cells proliferation, mesangial matrix broadening;3The expression of AT1mRNA were increased obviously in localrenal of rats in model group, which suggest that the activation of localRAS system in renal may play a critical role in the progressive process ofIgAN. Our traditional Chinese medicine of supplementing qi, nourishingyin maybe achieved treatment effectiveness through blocking RASsystem;4AT1mRNA expression level and the synthesis of collagen type Ⅳwere positive correlation in rats of IgA nephropathy. This result suggestedthat the activation of local RAS system can stimulate synthesis ofextracellular matrix. Our traditional Chinese medicine of supplementingqi, nourishing yin can suppress this effect significantly to achieve thepurpose of delaying the progress of IgAN.

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CLC: > Medicine, health > Surgery > Urology ( urinary and reproductive system diseases) > Kidney disease > Nephritis
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