| Objective: Using the unilateral ureteral obstruction (UUO) to make arenal interstitial fibrosis (RIF) model, this paper aims to observe the change ofexpression levels of renal Vascular Endothelial Growth Factor (VEGF) andangiopoietin-1(Ang-1) as well as angiopoietin-2(Ang-2) in different UUOgroups, explore the effect of the Chinese traditional medicine (CTM)dedicated to activating blood circulation and dredging the collaterals on renalinterstitial micro-vessels and its action mechanism, with a view to provideexperimental evidence for clinical application.The breakage and loss of renal microvessels, particularly the breakageand loss of peritubular capillaries (PTC) has drawn great attention in recentyears during the treatment course of kidney diseases. As a common finding inpatients with progressive chronic renal disease of all types, RIF arises out of aclose connection with the breakage and loss of PTC. Exploring thedevelopment mechanism of RIF to prevent and delay RIF has become vital inthe treatment course of kidney disease. In this regard, research from theperspectives of PTC breakage and loss is of great significance. UUO is anoptimal model for RIF research, since it reveals the development course ofObstructive Nephropathy and renal fibrosis diseases. Therefore, we choose theUUO model to conduct this research.RIF is the common pathological foundation for various chronic kidneydisease (CKD) leading to the chronic renal failure (CRF). CKD and CRF bothbelong to the TCM syndrome scope of edema,consumptive disease andobstruction and rejection, etc. In the last two years, there are medical mastersholding the view that CKD is similar to the "mass in the abdomen" accordingto the excessive storage of ECM and the Fibroblast Proliferation of RIF syndromes. The Jin Gui Yao Lue-Edema Chapter put forward that theunsmooth blood circulation may lead to Edema, emphasizing the unsmoothblood circulation and the blood stasis play important roles in the formation ofhydrosphere diseases, which has also drawn concerns of the physicians ofsuccessive generations. Meanwhile, CKD, as a chronic long illness,"a longillness affects collateral circulation" and "a long illness leads to the stagnationof blood circulation" are among its basic pathogenesises. Accordingly,"promoting blood circulation by removing blood stasis" and "removing bloodstasis for promoting tissue regeneration" is an important TCM therapy fortreating CKD. Inspired by Jin Gui Yao Lue-ChapterⅥ:On Pulse SyndromeComplex And Treatment Of Arthralgia Due To Stagnation Of BloodConsumptive Diseases,Da Huang Zhe Chong Pill is dedicated to activatingblood circulation and dredging the collaterals and has become a commonclinical prescription for treating CKD.This research applies UUO to copy RIF model and applies Da Huang ZheChong Pill for treatment, in comparison with valsartan treatment, to observethe changes of such indicators of the UUO rats like VEGF, Ang-1and Ang-2,and to explore the effect of Da Huang Zhe Chong Pill on UUO spicatuson rats,for a purpose to provide theoretic evidence for the clinical application of DaHuang Zhe Chong Pill.Method:1Select50female SD rats of healthy and clean grade at weight of170±10g respectively.: After one week's adaptive feeding and the examinationresult of urinary protein negative, weigh each rate and label them. Dividingthem randomly into5groups with10rats for each group: Group S (Shampgroup, S), Group M (Model group, M), Group V (Valsartan group, V) andGroup DM (DHZCM group,DM) and Group DH (DHZCH group,DH). ExceptGroup S, the rats in other groups all adopt UUO to establish UUO rats Model.2Drug administration during experiment: the drug administration for ratsis calculated from the conversion formula between human drug administrationto experimental animal drug administration. Ordinary feeding is for all five groups, of which, distilled water10ml·kg-1·d-1is administered to the rats inGroup S and M respectively while10mg·kg-1·d-1Valsartan for Group V,3g·kg-1·d-1DHZCM solution to the rats in Group DM and6g·kg-1·d-1DHZCHsolution to the rats in Group DH. One drug administration for each day andsuch treatment lasts for14days. This experiment duration is2weeks in total.3Detection of Indicators: After weighing them one day prior to theending of the experiment, put the rats inside the metabolic cages and collect24h urine under the normal fasting and test the24h urine amount and24hproteinuria excretion. Collect blood samples at the end of experiment toexamine Scr and BUN and calculate the Ccr; collect the some tissue from theleft kidney and place it quickly into the4%paraformaldehyde solution,dehydrate and embed the tissue in paraffin, cut into sections at thickness of3μm. After HE staining and Masson trichrome staining, observe by lightmicroscopy the structure of in renal tubulointerstitium part as well as theexpression of VEGF, Ang-1and Ang-2.Result:1General information and24h proteinuria excretion examination results.There is no significant change on the mentality, hair, activities, diets, drinkingof rats in Group S; the rats in Group M are sparsely and disorderly coated withdimming coat color, lack of appetite and in torpor; the rats in Group V, GroupDM and Group DH improve in the overall conditions of diets, weights andactivities. There is no apparent differentiation (P>0.05) among the24hproteinuria excretion.2Examination results of renal functions of ratsBUN examinations results: In comparison with Group S, the BUN levelsof Group M rise obviously (P<0.01);In comparison with Group M, there issignificant difference (P<0.01for all) among the decline of BUN level ofGroup V, Group DM and Group DH. In comparison with Group V, here issignificant difference (P<0.01for all) between the decline of BUN level ofGroup DM and Group DH. However, there is no significant difference(P>0.05) between Group DM and Group DH. Scr Examination Results：In comparison with Group S, the Scr level ofGroup M rises obviously (P<0.01). In comparison with Group M, there issignificant difference (P<0.01for all) among the decline of Scr level of GroupV, Group DM and Group DH. In comparison with Group V, there is significantdifference (P<0.01for all) between the decline of Scr level of Group DM andGroup DH. There is also significant difference (P<0.01) between Group DMand Group DH.There is no significant difference (P>0.05) among the Ccr levels of thegroups.3Pathomorphological observation of renal tissue by light microscopyPathological changes on the renal tissue is not found in Group S; adrenalcortex become thinner and renal tubular epithelial cells bear swollen andnecrotic shape; some renal tubular focal atrophy with the expanded cavity andobvious increase of interstitial width can be observed. A dense inflammatoryinfiltrate can be observed in the interstitial cells. After Masson trichromestaining, the collagenous fibrocytes hyperplasied measurably in focal atrophycould be seen in the widened interstitial part; In comparison with Group DMand Group DH as well as Group V, the pathological changes on the renaltubular and renal interstitium improves obviously, specifically, the atrophyof renal tubular allieviates, the expansion of cavity slows, the necrotic cellsreduces and the widening of renal interstitium mitigates and the inflammatoryinfiltrate becomes less dense.4Expression of VEGF, Ang-1and Ang-2by immunohistochemistryVEGF examination result:VEGF is mainly located in the renal tubularcells and glomerular visceral epithelialcells in Group S.Besides theexpressions in the above part, there is a few VEGF expression in theglomerular mesangial cells and the VEGF expression of renal interstitiumdeclines significantly (P<0.05),the expression levels of Group V and GroupDH are obviously higher than that of Group M (P<0.01),there is no obviouslydifference between the Group DM and Group M.Ang-1is mainly located in the renal tubular cells in Group S.The Ang-1 expression of renal interstitium declines significantly (P<0.001),the expressionlevels of Group V,Group DM and Group DH are obviously higher than that ofGroup M (P<0.05,P<0.05,P<0.001),Group DH is better than Group V andGroup DM(P<0.05),there is no obviously difference between the Group V andGroup DM(P>0.05).Ang-2examination result:Ang-2is mainly located in the renal tubularcells in Group S.The Ang-2expression of renal interstitium declinessignificantly (P<0.01), the expression levels of Group V,Group DM andGroup DH are obviously higher than that of Group M (P<0.05),there is noobvious difference among the three groups(P>0.05).Conclusion:1Da Huang Zhe Chong Pill could reduce the RIF Scr level and Bun level ofthat are caused by UUO rats.2Da Huang Zhe Chong Pill could increase the expression levels of VEGF,Ang-1and Ang-2in renal tissues to protect the capillary endothelial cells ofrenal tuberla and reverse the broken angiogenesis, for a purpose to improveand inhibit RIF. |
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