Kidney Slow Down Side Of Apoe - / - Mice Renal Artery Atherosclerosis And The Study Of The Influence Of Hif

Ischemic renal disease which caused by atherosclerotic renal artery stenosis is an important etiology to lead to end stage renal disease.The key link of ARAS is "Hypoxia-Microvascular abandoned-Tubule interstitial fibrosis".Previous we have confirmed that the basic pathogenesis of ARAS is qixushiyuhuzu.We present Yiqihuoxuequshi treatment principles.Therefore,we come up with a Working hypothesis which is the affect of Yishenhuanshuai decoction on ApoE-/-mouse’s "Hypoxia-Microvascular abandoned-Tubule interstitial fibrosis".Use the ApoE-/-mouse which is recognized by Internationally to establish the model.Apply the light microscopy and fluorescence real-time quantitative PCR technology.Observe the affect of Yiqihuoxuequshi herbs on Renal artery,capillaries and tubulointerstitial fibrosis from Organization and gene multilevelly.Especially observe the affect of HIF-la,TGF-(3,VEGF and Flk-1.Searching the targets and mechanisms to treat the ARAS to lay the foundation for treating ARAS with traditional Chinese Medicine.Research purpose:This research further expand the range of treatment based on the previous research that the affect of Yiqihuoxuequshi on the structure and function of vascular endothelial function and kidney structure.Do the detailed research according to the key pathological link-"Hypoxia-Microvascular abandoned-Tubule interstitial fibrosis",by establishing the model of ApoE-/-mouse.So as to confirm that Yishenhuanshuai prevent the tubule-interstital fibrosis which through prevent the damage of the function of vascular endothelium.Research method:lAnimal experiments1.1Use the model of internationally recognized ApoE-/-mouse.Buy45Male C57BL-6J ApoE-/-mouse in the years of20weeks with standard of SPF,15Male C57BL/6J wild mice in the years of20weeks with standard of SPF,then feed with high fat diet.32weeks later the Tunica intima of artery appear hyperplasia on ApoE-/-mouse. Then Randomly divided into blank control group,herb group and Western medicine group. Herb group intragastric administration0.01mg/d.g with Yishenhuanshuai decotion which was obtained by Guanganmen Hospital preparation room,each ml containing crude drug extract1.6g.Western medicine group intragastric administration0.0267mg/d.g with Telmisartan.The blank group and grow wild group infusioned with physiological saline.1.2Interfere in administration thirteen weeks,mouse artery blood serum was separated to determine renal function (Scr and BUN)and blood fat(TG、LDL-C、VLDL-C).1.3After executing the mouse,use the renal artery specimen to observe the artery histology by HE dyeing(the renal artery stenosis and the condition of plaque). 1.4After executing the mouse,use the renal specimen to observe the renal histology by PAS dyeing(the evaluation of the damage of tubular-interstital).1.5Molecular biology detection:real-time PCR was used to determined HIF-la, TGF-β, VEGF, Flk-1expression of renal tissue.2Statistical AnalysisExperimental data were compared groups analysis of t using statistical software SPSS17.0,the result is expressed as (x±s). P<0.05have the statistical significance.Experimental Results:1Yishenhuanshuai decotion on ApoE-/-renal function and blood fat effects.Scr and BUN have statistical differences(P<0.05) between blank group and wild group.It shows renal function appear abnormal after modeling.Compare the blank group with the telmisartan group and herb group,it appears significant difference with Scr and BUN.It turns out Yishenhuanshuai and western medicine can both improve the renal function.The Scr and BUN between Yishenhuanshuai and western medicine also have differences(P<0.05).It shows Yishenhuanshuai improve more than western group for the renal function;The TG、 LDL-C and VLDL-C between blank group and wild group all have statistical significance (P<0.05).It shows mouse appearing significant dyslipidemia after modeling.TG> LDL-C、 VLDL-C have differences between the Yishenhuanshuai and blank group(P<0.05).It shows Yishenhuanshuai can significantly reduce the blood fat,so as to reduce the risk of atherosclerosis.Because the Western medicine telmisartan can not reduce the blood fat.So compare with the model group there have no statistical differences (P>0.05).2The Yishenhuanshuai decoction has an affect on the ApoE-/-mouse that have renal artery hurt and kidney damage.After visitioning, we can see the blank group mouse have less obvious tubular interstitial damage situation;Assess the damaged condition of artery refer to the Integral standard of Renal microvessel damage by kidney pathological study committee.The results shows: Observe under the light microscopy,renal artery of model group appears significant stenosis,the subendothelial have plenty of foam cells,even come severe cases appears fibrous plaque and fibrous cap.The degree of Renal arterial lumen’s stenosis significant lighten.The luminal stenosis of westen group lighten.The statistical analysis shows:Blank group and wild group are different (P<0.05).It appears significant arterial lumen damage after modeling.It have significant statistical differences with Yishenhuanshuai decoction and western medicine (P<0.05),it means both medicine can reduce the renal artery injury.Yishenhuanshuai decocion and western medicine have statistical differences (P<0.05).It means compared with western medicine,Yishenhuanshuai decoction is a better choice to alleviate the renal artery injury.3Yishenhuanshuai decoction on TGF-pmRNA、HIF-1αmRNA、Flk-1mRNA、VEGFmRNA expression.Using the real-time PCR method(act the GAPDH as inner parameter)determine the TGF-PmRNA、HIF-1αmRNA、Flk-1mRNA、VEGFmRNA.Though the diagram of curves,we can observe blank group can obviously advange the level of the TGF-βmRNA、HIF-1αmRNA which have the function of profibrotic,it also can down regulate the level of Flk-1mRNA、 VEGFmRNA which have the function of anti-profibrotic.Yishenhuanshuai decoction and western group can both down regulate the level of TGF-βmRNA、HIF-1αmRNA,and up regulate the level of Flk-1mRNA、VEGFmRNA.Research conclusion:lYishenhuanshuai decoction can obviously improve the function of renal disease,decrease the Scr and BUN;The affect of blood fat:there has no affect on TG,but has obsviously improving on LDL-C and VLDL-C.2Yishenhuanshuai decoction can improve the addition of renal artery damage.3Yishenhuanshuai decoction can down regulate the level of TGF-βmRNA、HIF-1αmRNA,and up regulate the level of Flk-1mRNA、VEGFmRNA.