水蛭素调节断指再植大鼠皮瓣成活、血管生成、炎症因子表达的作用

Effect of hirudin on survival, angiogenesis and expression of inflammatory factors in replanted flaps of rats with finger replantation

目的探讨水蛭素调节断指再植大鼠皮瓣成活、血管生成、炎症因子表达的作用。方法选取80只Wistar大鼠为研究对象,建立断指再植模型后随机分为两组:模型组、水蛭素组,均40只。水蛭素组大鼠皮瓣局部sc天然水蛭素6 ATU/次,2次/d,模型组大鼠sc生理盐水。术后7 d,观察大鼠皮瓣存活率;制备皮瓣组织匀浆,提取总蛋白,采用ELISA双抗体夹心法测定血管内皮生长因子(VEGF)、白细胞介素-6(IL-6)、细胞间黏附分子-1(ICAM-1)、肿瘤坏死因子-α(TNF-α)表达情况;采用免疫印迹法测定皮瓣中细胞外调节蛋白激酶(ERK1/2)、Bcl-2、p38丝裂原活化蛋白激酶(p38 MAPK)表达;HE染色法测定皮瓣中炎症介质浸润情况;CD34免疫组织化学染色法检测血管密度值。结果水蛭素组大鼠皮瓣存活率、血管密度值、VEGF水平、ERK1/2相对表达量均明显高于模型组,而p38 MAPK、Bcl-2相对表达量,IL-6、ICAM-1、TNF-α水平明显低于模型组,差异均具有统计学意义(P<0.05)。结论水蛭素在断指再植模型大鼠中具有提高皮瓣成活率、促进VEGF表达、抑制细胞凋亡、减少炎症因子表达作用。

Objective To investigate the effect of hirudin on survival, angiogenesis and expression of inflammatory factors in replanted flaps of rats with finger replantation. Methods Totally 80 Wistar rates were selected to establish finger replantation model, and they were randomly divided into two groups with 40 cases in each group. Hirudin group was given local subdermal injection of six ATU natural hirudin, twice a day, and control group was given local subdermal injection of normal saline. After 7 d, rat skin flap survival rate was observed; Flap tissue homogenate was prepared to extract total protein, and then detect vascular endothelial growth factor（VEGF）, interleukin-6（IL-6）, intercellular adhesion molecule-1（ICAM-1）, and tumor necrosis factor alpha（TNF-α） expression by using enzyme-linked immunosorbent assay（ELISA） double antibody sandwich method. Western blotting method was used for the determination of flap in extracellular regulated protein kinase（ERK1/2）, Bcl-2 and p38 mitogen activated protein kinase（p38 MAPK） expression; Flap inflammatory infiltration was determined by HE staining; CD34 immunohistochemical staining method was used to detect vascular density. Results The flap survival rate, vessel density, VEGF level, and relative expression of ERK1/2 of hirudin group were significantly higher than those of model group, but p38 MAPK and relative expression of Bcl-2 were significantly lower than control group（P〈0.05）; The IL-6, ICAM-1, TNF-α levels of hirudin group were significantly lower than those of control group（P〈0.05）. Conclusion Hirudin for rats model with finger replantation can increase flap survival rate, promote VEGF expression, inhibit cell apoptosis, and reduce inflammatory factors expression.