| Objective:To study the effects of Tempol(a new type small molecular oxygen free scavenger)on the survival of the LRPSF in rats;From anti-oxidative stress,promoting angiogenesis,reducing inflammatory reaction to explore its mechanism in three aspects,providing a theoretical basis for the pathogenesis of distal flap necrosis,clarifying the mechanism of ischemia reperfusion injury which caused the distal flap necrosis,providing new ideas in improving skin flap application efficiency and covering key wound.Method:1.84 male SD rats were randomly divided into normal saline control group and Tempol group,with 42 rats in each group.All the flaps were divided into 9cm*3cm based on Mcfarlane flap.In Tempol group from 15 min before operation to seventh days after operation,rats were administered Tempol(1:10 diluted with normal saline)according to daily intraperitoneal injection,at the same time points in the saline control group were injected with the same volume of saline.We observed the rats and skin flap survival conditions afer operation at different time points;The skin flap survival rate was measured,meanwhile perfused the blood vessels with Geltin lead oxide(1g/ml),then scan the vessels with MicroCT,and reconstructed the blood vessels’ three-dimensional imaging with 3D-viewer software,and analyzed the vascular structure,vascular volume and total length of blood vessels with MAT-LAB software after 7 days.2.Afer 7 days,The center of the flap by HE staining was used to observe the inflammation,hierarchical structure,angiogenisis,and tissue necrosis,similarily the immunohistochemical staining was used to observe the expression of the VEGF.3.Immediately after operation and 1,3,7 days,make the center of the flap tissue into homogenate,then use WST-1 method for antioxidant enzyme SOD and lipid peroxidation product MDA contents detecting;Inflammatory cytokines TNF-α and IL-6 were detected in skin tissue by using ELISA kit respectively.4.Result :1.All the experimental animals survived to the end of the experiment,each flap had no infection,hematoma and effusion.With the extension of time,the two groups had different degrees of distal flap necrosis,but the skin flap necrosis of Tempol group is lighter than that of the control group;The vascular distribution and continuity of Tempol group were better than the saline control group.Seventh days after operation,the vascular volume,the flap survival rate and the total vessel length in Tempol group were significantly higher than those in the saline control group(p<0.05 or p<0.01).2.Seventh days after operation,compared with the control group,histological and immunohistochemical staining showed that the skin layers of tissue structure is clearer,micro angiogenesis density was significantly increased,tissue inflammation and inflammatory cell infiltration is lighter,dermal tissue VEGF protein staining degree was obviously increased in the Tempol group.3.The flap homogenate of the two groups were mesured immediately after operation and 1,3,7 days,SOD,MDA,IL-6,TNF-α contents were statistically significant diferent between two groups afer 1,3,7days,in the Tempol gorup MDA,,IL-6,TNF-α were significantly lower than the saline control group,SOD was significantly higher than that in the saline control group(p<0.05);While there was no significant difference of these indexes in two groups immediately after operation(p>0.05).Conclusion :Pretreatment of Tempol resulted in a higher viability of random skin flap by reducing inflammation reaction and soft tissue edma,by increasing the expression of the VEGF protein,promoting microvessels vasculogenesis,by increasing SOD activity and reducing MDA,IL-6,TNF-α content,and by alleviating ischemia reperfusion injury in brief. |
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