局部移植人脐带间充质干细胞治疗大鼠皮瓣缺血再灌注损伤

摘要

目的 探讨人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,HUGMSCs)在受体内分化为血管内皮细胞、治疗皮瓣缺血再灌注损伤从而促进皮瓣成活的可能性.方法 体外增殖培养HUC-MSCs,并用流式细胞技术鉴定,以5-乙炔基-2’脱氧尿嘧啶核苷(EdU)标记HUC-MSCs后,移植于皮瓣缺血再灌注损伤的大鼠腹部皮瓣术区局部(治疗组),设PBS为对照组.术后每天观察皮瓣的颜色、皮纹、厚度、毛发生长、坏死范围及针刺出血情况等,并于术后7d处死大鼠,比较2组皮瓣的成活率,切取皮瓣组织行常规病理组织切片,分别进行免疫组织化学染色检测血管内皮生长因子(VEGF)的表达,EdU染色检测供体细胞在受体皮瓣组织内的分布或分化.结果 术后7d治疗组大鼠皮瓣成活率为(97.58±3.41)％,对照组为(54.37±8.78)％,治疗组高于对照组(P＜0.05).治疗组大鼠VEGF的表达密度为138.27±8.67,明显高于对照组的56.17±14.13(P＜0.05).在成活皮瓣的部分小血管内皮可见EdU阳性细胞连续分布.结论 HUC-MSCs在体内可以分化为血管内皮细胞,直接参与生成新血管,建立新的微循环,并能增加皮瓣局部VEGF的表达,促进血管形成,修复缺血再灌注损伤的皮瓣,减轻皮瓣坏死,促进皮瓣成活.%Objective To explore the possibilities of human umbilical cord mesenchymal stem cells (HUC-MSCs) differentiate into vascular endothelial cells,thus to treat flap ischemic reperfusion injury and then promote the flap survival. Methods HUC-MSCs in vitro,and flow cytometric identification.Labeling stem cells with EdU,and then transplant stem cells into local of ischemiareperfusion injury flap of rats( treatment group),setting PBS control group.Observe the flap survival rate 7 days after,and execute rats.Take flap tissues for pathological sections,and detect vascular endothelial growth factor (VEGF) by immunohistochemical staining,detect the distribution or differentiation of the donor cells in cell receptor by EdU fluorescent staining.Results 7 days after,the flap survival rate of treatment group was ( 97.58 ± 3.41 ) ％,the flap survival rate of control group was (54.37 ± 8.78 ) ％,the rate of treatment group was higher than control group(P ＜0.05).The density of micrangium and VEGF of treatment group was higher than control group obviously ( P ＜ 0.05 ).The continuous distribution of positive cells that labeled with EdU were found in new vascular endothelial of treatment group' s survival flap tissues.Conclusions HUC-MSCs could differentiate into vascular endothelial cells,directly involved in generating new blood vesselsand,additionally could increase expression of VEGF to promote the formation of new blood vessels,then to set up the new micro circulation,accordingly to treat flap ischemic reperfusion injury and then promote the flap survival.