Effect Of Improved Autologous Blood Treatment On Oxygen Carrying Capacity Of Erythrocyte And Wound Healing In Diabetic Mice

Objective:To study the effect of autologous blood transfusion treated with improved preservative solution on erythrocyte function and wound healing in diabetic mice.Methods:Thirty healthy adult male Kunming mice aged 8 weeks and weighing 26-30 g were selected.A diabetic mouse model was established by intraperitoneal injection of streptozotocin at 70 mg/kg body weight for 5 days after adaptive feeding for one week,When the blood sugar level of caudal vein was higher than 16.7 mmol/L,diabetic mice were considered to be successful in modeling.Successful diabetic mice were randomly divided into blood donation group and experimental group.The experimental group was randomly divided into standard group and improved group with 6 mice in each group.Diabetic wound healing model was made on the back of mice in the experimental group.Blood of mice in the in experimental group was collected by tail vein and placed in a pre-disinfected centrifugal tube containing anticoagulant and then preserved for 7,14,21 and 28 days with improved and standard blood preservation solutions,respectively.Detection of FHb in autologous blood of two groups by spectrophotometer.The H⁺,oxygen partial pressure at 50%oxygen saturation(P₅₀)and red blood cell deformability?RCD?in the improved blood preservation solution and standard blood preservation solution were assessed using the COBAS-B-123 Blood Gas and Electrolyte analysis system.ELISA was applied in order to detect Erythrocyte 2,3-DPG.At the same time,the platelet aggregation rate of autologous blood preserved for 7 days was detected by flow cytometry,and the blood sugar?Glu?,glycosylated hemoglobin?GHb?of autologous blood preserved by two groups of preservation solutions were detected.The white blood cell content in blood was detected by automatic cell analyzer.The mice in the improvement group were transfused with improved blood preservation solution?7 d?using autologous transfusion device on day 1 after the operation.The mice in standard group were transfused with standard blood preservation solution?7 d?using autologous transfusion device on day 1 after the operation.The wound healing of two groups of mice was observed on 0d,1d,4d,7d and 14d,and the mice were killed on14d.The degree of fibrosis was analyzed by means of HE,Masson staining methods.Results:Compared with the standard group,the improvement group exhibited decreased FHb content,and increased P₅₀ oxygen pressure,blood pH,2,3-DPG and RCD?P<0.05?.The platelet aggregation rate in the improved group was significantly lower than that in the standard group?P<0.05?.The levels of Glu,GHb and the number of WBC in the improved group were significantly lower than those in the standard group?P<0.05?.HE staining results revealed that compared with the standard group,the area of granular tissue on the skin and the vascular area were increased on the 14th day after operation in the improvement group.Masson staining revealed that,the expression of collagen fibers and the degree of fibrosis in the mice skin of the improvement group were elevated at 14 days after the operation,when compared with the standard group.Conclusion:The function of erythrocyte in autologous blood stored in two groups for 7 days was better than that of autologous blood stored for 14,21 and 28 days.The oxygen carrying capacity of autologous red blood cells preserved in modified solution was better than that of autologous blood preserved in standard preservation solution,and the wound healing of diabetic mice could be improved by transfusion of autologous blood preserved in modified solution.Improved preservation solution can promote wound healing in diabetic mice by improving the oxygen carrying capacity of red blood cells.