| Objective: To study the dynamic expression of adherence factorCD11b/CD18and ICAM-1in avulsed wound of rat skin by usingrecombinant hirudin locally, thus to explore the relationship amongrecombinant hirudin, the protection of avulsed wound of skin and thethree indexes above, and the effect and mechanism of injecting hirudinlocally on the survival rate of skin flap in avulsed wound.Methods:132SD male rats were divided randomly into blank controlgroup (60rats), hirudin local injection experimental group (60rats) andobservation group (12rats), and leaving4rats in blank control group andhirudin local injection group at each time to observe the survival of skinflap. Use the animal model studied by Wang Zhongtang and GuoShuzhong in2008, and we confirmed that making the model of skinavulsed wound with rats is feasible. After successful preliminaryexperiment, we designed any skin flap with pedicle at lateral abdomen(6cm×4cm), given8kg traction for8seconds, and connected withprecision instrument after fixation. After the skin was avulsed, dealt withit slightly and had it sutured in situ. The skin flaps of experimental groupwere divided into ten parts and injected hirudin40IU (1ml) locally. Theskin flaps of blank control group were also divided into ten parts andinjected with normal saline of equivalence (1ml). No treatment was dealt in normal observation group, extracting peripheral blood and skin todetect. Extracting peripheral blood at0h,2h,4h,5h,6h and12h ofexperimental group and control group, using flow cytometry to detect thepositive expression rate of CD11b/CD18in the cytoplasm of neutrophilicgranulocyte, and detected the ICAM-1expression of the surfaceendothelial cell by immunohistochemistry decoration. Makingquantitative analysis to comprehend the dynamic changes of the threeindexes, at last observed the healing of skin flap and calculate survivalrate of avulsed skin flap.Results: By observing the changes in experimental group and blankcontrol group and comparing each index at each time after surgery, wefound that there was hyperemia and edema in skin flap of two groups.The skin flap of control group showed different extent of congestion threedays after surgery, with a dividing line between black crust and livingtissue. Stripping black crusts, there was blood exudation and whitesecretion, with granulation tissue growing inside. At the meantime, thecongestion and edema of skin flap had alleviated a bit, no flare andeffusion at the fringe of skin flap three days after surgery in experimentalgroup, and after six days, the skin flap survived fine, no effusion. The rateof survival area is90.74±1.52%and75.38±2.64%in experimental groupand control group, and there’s a significant difference between them(p<0.01). The positive expression rate of adherence factor CD11b/CD18 in blood after avulsion increased gradually as time went by, and reachedthe peak in12h. By quantitative statistics, we found that at0h, thepositive expression rate of experimental group and control group is24.13±2.36and24.75±3.06respectively, and there was no significantdifference between them (p>0.05). The positive expression rate ofexperimental group and control group is29.08±1.06and36.35±5.28(p<0.05)) at the2h time. As time went by, the positive expression rate of4h,5h,6h and12h is35.07±2.02and40.08±1.30,39.65±1.31and44.87±5.57,45.52±3.39and52.48±5.85,52.53±3.16and71.32±3.57, andthere was significant difference between each of them(p<0.01). As timeprolonged, the positive expression of CD11b/CD18of experimental andcontrol group both raised, however, the experimental group wassignificantly lower than control group (p<0.05). The moment skin wasavulsed, the ICAM-1could be detected in experimental group and controlgroup, the expression data is5.31±2.12and5.25±1.87, and no significantdifference between them. As time passed by, the positive expression rateof ICAM-1upgraded gradually, at2h,4h,5h,6h and12h, the results are8.34±1.57and5.83±3.21,12.35±2.36and7.96±3.57,17.87±2.47and10.03±2.75,21.16±3.06and12.37±2.84,25.35±3.56and17.36±2.32respectively. Of the total, the positive expression rate of blank controlgroup was significantly higher than experimental group (p<0.01), whichindicated that compared with control group, the positive expression of adherence factor CD11b/CD18and ICAM-1lowered obviously. Inaddition, the healing rate of experimental group is elevated significantlycompared with control group.Conclusion: To inject recombinant hirudin locally may effectivelyrelieve the effect of adherence factor CD11b/CD18in blood and ICAM-1in tissue, alleviate inflammatory reaction, and improve the endothelialcell injury in the process of ischemia-reperfusion, thus to increase thehealing rate of skin flap after avulsed wound significantly. |
PDF Full Text Request