| IntroductionThe incidence rate of arteriosclerosis obliterans(Arteriosclerosis Obliterans,ASO) is increasing at an alarming rate year by year,ranks the first place of the lower limb ischemia diseases.And,the atherosclerosis(atherosclerosis,As) is the main pathological basis of blood vessel system diseases,arteriosclerosis will cause artery wall thickening,hardening and lose flexibility,which could ultimately lead to vascular stenosis and occlusion.If we did not pay attention to those,the patient would feeling chilly,numbness and pain of the limbs,which have big effect on the patient's quality of life;the more seriously that will results in gangrene,amputation and even lead to death. The pathogenesis of arteriosclerosis obliterans(ASO) is not clear,thus the establishment of the arteriosclerosis obliterans(ASO) pathological animal model has significant importance to proven the etiology,pathophysiology and pathogenesis of ASO,to research and develop the drugs which could prevent ASO.Nitric Oxide is a molecules which autocaraine by the endothelial itself.It plays important role in maintaning the vessel tension,regulating blood pressure,inhibting the migration and proliferation of vascular smooth muscle cells,inhibiting the aggregation the platelet and the leucocyte adhereing to artery wall.NOS,as a determinant kinase of NO synthisis,has important effect in vivo.NOS distribute very broadly invivo.The eNOS is the most important.We first inject distilled water in rats femoral-popliteal artery,superficial damage of the arterial intima is caused by low permeability, non-mechanically,then feed as high-fat,high-cholesterol diet,and establish a mature, short-cycle experimental period of arteriosclerosis obliterans model of ratsThen we detect the expression of eNOS in the rat model of arteriosclerosis obliterans with real time PCR.These all have great effect on uncovering the etiology of ASO and seeking new target for gene therapy.Materials and methods Experimental animals and materialsAnimals:health purebred 70 male Wistar rats,weight 200 g(3-4 months).High-fat diet:62.8%basis feeds+20%lard feeds+15%cholesterol+2% sodiumcholate+0.2%Propylthiouracil+vitamin D 2.Identification of the model of hindlimb arteriosclerosis obliteransGet the injury artery 15,30 and 90 days after surgery.Extract the NOSs mRNA,Sythesize cDNA with RT-PCR.Compare the expression of eNOS mRNA in injury artery with real-time PCR which conducted with SYBR-GREENâ… .Result1.All of the surgery successful completed.2.The rat hindlimb arteriosclerosis obliterans model was successfully established.Three months later,the high-fat diet+injury group(group D) showed typical ASO pathological changes at optical microscope observation,endothelial cell loss,internal elastic plate injuried(interruption or loss),structural anomalies,intima hypertrophy and hyperplasia of medial smooth muscle,through the internal elastic plate,migrate to the subendothelium,Connective tissue proliferated and arranged disorder,lumens stenosis, thrombosis and foam cells can be seen in plaques,also some visible necrosis and calcification foci formation were observed.Establish the mature,short-cycle experimental period of arteriosclerosis obliterans model of rats,similar to the ASO of human.3.The expression of Nos3 gene in the injury artery.The result of real-time PCR show that the expression of Nos3 is respectively 0.7219,0.4444,0.4601,lower compare with control group as 1.There is significant difference.This indicate that the expression of Nos3 was down-regulated in the injuried artery.ConclusionInjection of distilled water into rats femoral-popliteal artery can cause endothelial injury,and feed the high-fat,high-cholesterol diet can establish the mature, shortcycle experimental period of arteriosclerosis obliterans model of rats,similar to the ASO of human.The expression of Nos3 was down-regulated in the injuried artery.
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