The Research Of OECs Transplantation Modified With EsRAGE Gene In The Treatment Of Diabetic Foot Ulcer

Objective:1. Making observations on healthy control group, simplex diabetes, diabeticischemic foot ulcers diabetic ischemic foot ulcers group, injection withouttransfection OECs Group, ischemic foot ulcers with diabetes mellitus after injectionof transfection of OECs Group, ischemic foot ulcers diabetic injecting DMEMCulture media group6Set of muscle tissues in rats RAGE,NF-κB,VEGF,TGF-Beta1mRNAExpression and protein levels.2. Studying on the treatment of ischemic diabetic foot ulcers of esRAGE genetransfection in OECs cells while improving the local esRAGE standards.Methods:Preparing60SD male rats, weight260-300g, for the research, selecting10randomly as normal control group rats (N Group), and setting the rest50rats asmodel of diabetes. Selecting10rats as simple diabetes group (DM) randomly withinthe rest50rat model of diabetes mellitus, the remaining40in diabetic rats will beligated the left lower extremity blood vessels, dividing them into: foot ulcers diabeticischemic Group (DFU),10rats; ischemic foot ulcers diabetic group injected nottransfection OECs Group (DFU+OECs groups)10rats; ischemic foot ulcers diabeticgroup after injection of transfection of OECs Group (DFU+esRAGE+OECsGroup)10rats; and Ischemic foot ulcers diabetic group injected DMEM culture fluidGroup (DFU+DMEM),10rats. Diabetes group in rats with intraperitoneal injectionof1%effect of Streptozotocin (according to the weight50mg/kg), the control groupinjected doses of citric acid-sodium citrate buffer. Those which are fasting bloodglucose (FBG)≥16.7mmol/l will be considered as successful diabetic model. After aweek, each one will be injected with10%chloral hydrate3.0ml/g k intraperitoneal inanesthetic, along the left groin to medial to the midpoint of about1.5cm longincisions, Isolate the femoral artery and its branches and ligation, ligation of thepopliteal artery and its branches simultaneously. DFU+OECs Group, DFU+esRAGE+OECs Group for cell transplantation, DFU+DMEM group using the same method of injection DMEM culture fluid were studied. Femoral-popliteal artery to theadductor to gastrocnemius select6points, every2mm anesthetic liquid syringe needleintramuscular transplantation50uL, a total of0.3mL(total number of cells5x105).The DFU+OECs Group which be injected would not transfected with OECs,DFU+esRAGE+OECs Group transfected with OECs after injection, DFU+DMEMGroup injected DMEM culture fluid. Meanwhile, in the left dorsal foot, causingpenetration of lower limb full thickness dermal wounds, cut off skin5mm*5mm.Transplanted rats reared under the same conditions after4weeks, while observing thewounds to heal. At the end of fourth week, killing those rats with dislocated cervicalvertebrae, and transplanting the cells in the muscle tissue. Detect the capillaryendothelial cell proliferation and distribution density by Haematoxylin and eosin. UseTUNEL method to detect the expression of cell apoptosis. Test the mRNA and proteinexpression levels of RAGE, and NF-κband the VEGF and TGF-β1by usingfluorescence quantitative PCR and Immunohistochemistry.Results: With DFU, DFU+DMEM group NF-kB expression in rat musclestrength, DFU+OECs, DFU+esRAGE OECs group muscle tissue of rats nf-kappa Bexpression is reduced, and the difference was statistically significant (P<0.01);Compared with DFU+OECs group, DFU+OECs+esRAGE group rats the nf-kappa Bexpression to decrease, the difference was statistically significant (P<0.01).With DFU, DFU+DMEM group RAGE expression in rat muscle strength, DFU+OECs, DFU+esRAGE OECs group muscle tissue of rats RAGE expression isreduced, and the difference was statistically significant (P<0.01); Compared with theDFU+OECs group, DFU+OECs+esRAGE group rats reduced the volume of RAGEexpression, the difference was statistically significant (P<0.01).With DFU, DFU+DMEM group TGF-beta1expression in rat muscle strength,DFU+OECs, DFU+esRAGE OECs group rats increased muscle tissue expression ofTGF-beta1, and the difference was statistically significant (P<0.01); Compared withthe DFU+OECs group, DFU+OECs+esRAGE group rat TGF-beta1expressionquantity increases, the difference was statistically significant (P<0.01).With DFU, DFU+DMEM group, DFU+OECs group, compared to three groupsof DFU+OECs+esRAGE group rats VEGF protein expression quantity increases slightly, the difference was statistically significant (P <0.01).With DFU group, DFU+DMEM cell apoptosis in rat muscle tissue volume,DFU+OECs, DFU+esRAGE+OECs group rats to reduce muscle tissue apoptosis,with the difference was statistically significant (P<0.01); DFU group, DFU+DMEMcell apoptosis in rat muscle tissue volume increased in rats in the normal controlgroup and DM group (P<0.01); Compared with the DFU+OECs group, DFU+OECs+esRAGE group rats decreased apoptosis and the difference was statisticallysignificant (P <0.01);With DFU,DFU+DMEM group RAGEmRNA expression in rat musclestrength,DFU+OECs,DFU+esRAGE OECs group muscle tissue of rats RAGEmRNAexpression increased, and the difference was statistically significant (P<0.01);Compared with the DFU+OECs group, DFU+OECs+esRAGE group rats decreasedRAGEmRNA expression, and the difference was statistically significant (P<0.05);Conclusion:1. esRAGE genetic modification of OECs transplantation in the rat muscle tissueRAGE, NF-kB protein expression and RAGEmRNA levels, apoptosis of rats werelower than that of ischemic diabetic foot ulcers.2. esRAGE genetic modification of OECs transplantation in the rat muscle tissueTGF-beta1, VEGF protein expression were higher than in ischemic diabetic footulcer in rats.3. esRAGE genetic modification of OECs transplantation in diabetic ratsischemic foot ulcer repair.