局部应用人脐带间充质干细胞治疗皮瓣缺血再灌注损伤的实验研究

发布时间：2018-03-10 00:17

本文选题：人脐带间充质干细胞　切入点：外科皮瓣　出处：《青岛大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的探讨人脐带间充质干细胞(Human umbilical cord mesenchymal stem cells, HUC MSCs)在受体内分化为血管内皮细胞,从而治疗皮瓣缺血再灌注损伤并且促进皮瓣成活的可能性。方法体外增殖培养HUC MSCs,并用流式技术鉴定,以EdU(5-乙炔基-2’脱氧尿嘧啶核苷,5-Ethynyl-2'-deoxyuridine, 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注射对照组。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,比较两组皮瓣的成活率,并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及分化。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(P0.05)。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(P0.05)。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。
[Abstract]:Objective to investigate the possibility of differentiation of human umbilical cord mesenchymal stem cells (HUC MSCs) into vascular endothelial cells (VECs) in order to treat ischemic reperfusion injury and promote the survival of human umbilical cord mesenchymal stem cells (HUC MSCs). Methods HUC cells were cultured in vitro and identified by flow cytometry. HUC MSCs was labeled with Edu 5-ethynyl-2'-deoxyuridine (Edu) -5-ethynyl-2'deoxyuridine (Edu). The local area of abdominal flap was transplanted to the rats with ischemia reperfusion injury (treatment group, PBS injection control group). The color, skin stripe, thickness, hair growth, necrotic range and acupuncture bleeding were observed every day after operation. The rats were killed on the 7th day after operation. The survival rate of the flap was compared between the two groups. The expression of vascular endothelial growth factor (VEGF) was detected by immunohistochemical staining and Edu staining was used to detect the distribution and differentiation of donor cells in the recipient flaps. Results the survival rate of flap was 97.58 卤3.41 in the treatment group and 54.37 卤8.78 in the control group. The expression density of VEGF in the treatment group was 138.27 卤8.67, which was significantly higher than that in the control group (56.17 卤14.13P0.05. EdU). Positive cells were continuously distributed. Conclusion HUC MSCs can differentiate into vascular endothelial cells in vivo, participate directly in the formation of new vessels, establish new microcirculation, increase the expression of local VEGF, promote angiogenesis, and repair the flap with ischemia reperfusion injury. Reduce the necrosis of the flap and promote the survival of the flap.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329

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