水蛭素对大鼠缺血皮瓣血管生成的促进作用及机制  

The mechanism of hirudin promoting angiogenesis of ischemic flap in rats

作　　者：高栋梁[1] 张雷[1] 王亚康 薛宏斌[1] 杨喜明[1] 郭建斌 侯国玲[1] GAO Dongliang;ZHANG Lei;WANG Yakang;XUE Hongbin;YANG Ximing;GUO Jianbin;HOU Guoling(Department of Burn and Plastic Surgery, The Affiliated Hospital of Yan'an University, Yan'an 716000, Shanxi, China;Department of Hip Surgery, Honghui Hospital, Xi'an Jiaotong University, Xi'an 710054, China)

机构地区：[1]延安大学附属医院烧伤整形外科,陕西延安716000 [2]西安交通大学附属红会医院髋关节外科,陕西西安710054

出　　处：《西部医学》2020年第11期1584-1588,共5页Medical Journal of West China

基　　金：陕西省科学技术厅科研课题(2019JQ-536)。

摘　　要：目的研究水蛭素对大鼠缺血皮瓣血管生成的促进作用及其可能机制。方法取10只SPF级SD大鼠,分别于大鼠背部中线双侧制备缺血皮瓣模型,右侧皮下注射水蛭素2 ATU(实验组),左侧皮下注射等量生理盐水(对照组)。分别于术后3、7 d,对皮瓣组织行HE染色,经免疫组织化学染色检测后计算血管密度值;术后7 d时,计算皮瓣成活率,用蛋白质免疫印迹法检测血管内皮细胞生长因子(VEGF)、血小板反应蛋白1(TSP-l)、p38 MAPK及ERK1/2磷酸化表达情况。结果HE染色显示,实验组术后3、7 d时皮瓣组织坏死较对照组明显减少,新生血管较对照组明显增多(P<0.05)。与对照组比较,实验组术后7 d时皮瓣成活率明显增高(P<0.01)。免疫组织化学染色结果显示,术后3 d对照组皮瓣组织表皮层、真皮层均出现明显坏死;术后7 d时,由于对照组皮瓣组织大面积坏死,未观察到成功染色的血管;实验组术后3、7 d时皮瓣组织均未见明显坏死区。实验组术后3、7 d时血管密度值较对照组明显增加(P<0.01),实验术后7 d时血管密度值明显高于术后3 d(P<0.05)。实验组术后7 d时VEGF表达水平较对照组明显升高,TSP-1表达水平较对照组明显降低(P<0.01)。与对照组比较,实验组术后7 d时p38 MAPK磷酸化表达水平明显降低,ERK1/2磷酸化表达水平明显升高(P<0.05)。结论水蛭素具有促进大鼠缺血皮瓣新生血管形成的作用,其作用机制可能在于诱导血管内皮生长促进因子VEGF的表达和下调血管内皮生长抑制因子TSP-1的表达,可能与p38 MAPK、ERK1/2信号通路密切相关。Objective To study the effect of hirudin on angiogenesis of ischemic flap in rats and its possible mechanism.Methods 10 SD rats were used to establish the ischemic skin flap model.Hirudin with 2 ATU was injected subcutaneously on the right side(experimental group)and normal saline was injected subcutaneously on the left side(control group).On the 3rd and 7th day after operation,the flap tissues were examined by pathology,immunohistochemistry and vascular density.On the 7th day after operation,the survival rate of flap was calculated.The expression of vascular endothelial growth factor(VEGF),thrombospondin-l(TSP-1),p38 MAPK and ERK1/2 phosphorylation were detected by immunoblotting.Results Pathological examination showed that the necrosis of flap tissue in the experimental group was significantly reduced and neovascularization was significantly increased compared with the control group on the 3rd and 7th day after operation.Compared with the control group,the survival rate of flap in the experimental group was significantly increased 7 days after operation(P<0.01).The results of immunohistochemistry showed that the epidermis and dermis of the skin flap in the control group were obviously necrotic on the 3rd day after operation,and no blood vessels were observed because of the large area necrosis of the skin flap in the control group on the 7th day after operation,and no obvious necrosis area was found in the skin flap in the experimental group on the 3rd and 7th day after operation.The vascular density of the experimental group was significantly higher than that of the control group on the 3rd and 7th day after operation(P<0.01),and that of the experimental group on the 7th day was significantly higher than that on the 3rd day after operation(P<0.05).The expression level of VEGF and TSP-1 in the experimental group was significantly higher than that in the control group(P<0.01).Compared with the control group,the phosphorylation level of p38 MAPK in the experimental group was significantly lower and that of ERK1/2

关 键 词：缺血皮瓣 水蛭素 血管生成 血管内皮细胞生长因子 血小板反应蛋白1 

分 类 号：R628[医药卫生—整形外科]