Influences on wound healing in diabetic mice by blood transfusion treated with improved blood preservation solution
WEI Han-wei, ZHU Na-na, WANG Huan, LIU Xiao-qian, DUAN Li-shuang, ZHOU Xun, GUO Jian-rong
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Objective To study whether blood transfusion with improved blood preservation solution can improve wound healing in diabetic mice, and to investigate its possible mechanism. Methods Kunming mice were used to establish diabetic mice model. They were randomly divided into blood donation group and the experimental group. The blood was placed in sterile centrifugal tube containing anticoagulant and then preserved for 7, 14, 21 and 28 days with improved and standard blood preservation solutions, respectively. The oxygen carrying capacity and deformability of erythrocytes in each preservation period were tested, then the improved and standard autologous blood were retransfused with 7 days of preservation. The experimental group was randomly divided into the standard group and the improved group with 6 mice in each group.The wound healing of two groups of mice was observed on 0, 1, 4, 7 and 14 days, and the mice were killed on 14 days. The degree of fibrosis was analyzed by means of HE, Masson staining and immunohistochemistry methods. Phosphorylation of PI3K,AKT,MEK and ERK were analyzed by Western blot analysis. Results Compared with the standard group, the improvement group exhibited decreased fHb content, and increased P50 oxygen pressure, blood pH, 2,3-Diphosphoglycerate(2, 3-DPG) and red blood cell deformability(RCD)(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. Immunohistochemistry results indicated that in comparison to standard group, levels of HIF-1α, VEGF, and EGF in skin tissues of the improvement group increased (P＜0.05). The protein levels of PI3K, AKT, MEK,ERK and its phosphorylation increased significantly in the improvement group(P＜0.05). Conclusions Red blood cell treated with improved blood preservation solution may promote wound healing in diabetic mice by activating HIF-1α pathway.