转染leptin的胎盘间充质干细胞对放射损伤复合创伤愈合的促进作用

doi:10.13481/j.1671-587x.20160605

吉林大学学报(医学版) ›› 2016, Vol. 42 ›› Issue (06): 1066-1070.doi: 10.13481/j.1671-587x.20160605

转染leptin的胎盘间充质干细胞对放射损伤复合创伤愈合的促进作用

费倩怡¹, 刘书会¹, 朱振威², 邹永巍¹, 郑明¹, 刘春丽¹, 刘志辉³

1. 吉林大学口腔医院口腔颌面外科, 吉林长春 130021;
2. 吉林省长春市口腔医院, 吉林长春 130022;
3. 吉林大学口腔医院口腔修复科, 吉林长春 130021

收稿日期:2014-10-14 出版日期:2016-11-28 发布日期:2016-12-02
通讯作者: 刘春丽,教授,硕士研究生导师(Tel:0431-85579325,E-mail:iceapplelsh@hotmail.com);刘志辉,副教授,硕士研究生导师(Tel:0431-88796018,E-mail:liuzhihui1975@sina.com) E-mail:iceapplelsh@hotmail.com;liuzhihui1975@sina.com
作者简介:费倩怡(1986-),女,吉林省吉林市人,住院医师,医学硕士,主要从事整形与外科重建方面的研究。
基金资助:
吉林省发改委科研项目资助课题（2013C025-4）；吉林省自然科学基金资助课题（201015201）；吉林省卫计委青年基金资助课题（3D514CP23431）

Promotive effect of HPMSCs transfected with leptin on healing of combined radiation-wound injury

FEI Qianyi¹, LIU Shuhui¹, ZHU Zhenwei², ZOU Yongwei¹, ZHENG Ming¹, LIU Chunli¹, LIU Zhihui³

1. Department of Oral and Maxillofacial Surgery, Stomatology Hospital, Jilin University, Changchun 130021, China;
2. Changchun Stomatology Hospital, Jilin Province, Changchun 130022, China;
3. Department of Prosthodontics, Stomatology Hospital, Jilin University, Changchun 130021, China

Received:2014-10-14 Online:2016-11-28 Published:2016-12-02

摘要/Abstract

摘要：

目的：探讨瘦素（leptin）和人胎盘间充质干细胞（HPMSCs）对大鼠背部放射损伤复合创伤（放创复合伤）皮瓣愈合的促进作用，阐明其在放创复合伤愈合过程中的协同作用。方法：将30只雄性Wistar大鼠随机分为转染leptin的HPMSCs组（n=10）、转染空载体的HPMSCs组（n=10）和空白对照组（n=10）。建立放创复合伤大鼠模型，放疗后24 h各组大鼠局部皮下注射转染leptin的HPMSCs、转染空载体的HPMSCs以及生理盐水，观察各组大鼠皮瓣颜色、皮纹、质地、温度、毛发生长、渗出、坏死范围及针刺出血情况；检测各组大鼠皮瓣成活率；HE染色观察各组大鼠皮瓣组织形态表现；免疫组织化学法检测大鼠皮瓣中leptin和血管性血友病因子（vWF）的表达情况。结果：给药后第7天，各组大鼠皮瓣成活与坏死界限基本清楚，坏死区域皮瓣质地坚硬，成活区域皮瓣颜色红润，皮纹模糊，质地中等，转染leptin的HPMSCs组和转染空载体的HPMSCs组大鼠皮瓣坏死面积均小于空白对照组。给药后第7天，转染leptin的HPMSCs组大鼠皮瓣成活率高于转染空载体的HPMSCs组和空白对照组（P<0.05），转染空载体的HPMSCs组大鼠皮瓣成活率高于空白对照组（P<0.05）。给药后第14天，HE染色检测，转染leptin的HPMSCs组大鼠皮瓣肉芽组织最为丰富，新生毛细血管和成纤维细胞最多。给药后第14天，免疫组织化学染色，转染leptin的HPMSCs组和转染空载体HPMSCs组大鼠皮瓣真皮下方可见大量的黄染颗粒（阳性表达），转染leptin的HPMSCs组胞浆中阳性表达的leptin细胞最多。结论：在局部微环境作用下，HPMSCs可以分化为血管内皮细胞，促进新生血管的形成，HPMSCs与leptin可协同促进放创复合伤皮瓣的愈合，HPMSCs携带目的基因或单独应用均可明显提高放创复合伤皮瓣的成活率，但转染leptin的HPMSCs作用力更强。

关键词: 人胎盘间充质干细胞, 瘦素, 伤口愈合, 放创复合伤

Abstract:

Objective: To study the promotive effects of leptin and human placenta-derived mesenchymal stem cells(HPMSCs) on healing of combined radiation-wound injury of backs in the rats, and to clarify the synergistic effects of HPMSCs and leptin on healing of combined radiation-wound injury.Methods: A total of 30 Wistar rats were randomly divided into HPMSCs transfected with leptin group (n=10), HPMSCs group (n=10), and blank control group (n=10). The model of combined radiation-wound injury was constructed.The HPMSCs transfected with leptin,the HPMSCs transfected with empty vector, and normal saline were transplanted subcutaneously into the rats in various groups 24 h after radiotherapy, respectively. The color, dermatoglyph,hardness,temperature,haigrowth,exudation,necrotic aera coverage and acupuncture bleeding reaction of the rats in various groups were observed. The survival rates of flap of the rats in various groups was detected.The histomorphology of flap after radiotheraphy in various groups was observated by HE staining. The expressions of von Willebrand factor(vWF) and leptin in flap of the rats in various groups after radiotheraphy were detected by immunohistochemical method.Results: On the 7th day after administration, the boundary of survival area and necrotic area of the rats in various groups were clear.The nectotic area of flap was hard and the color was ruddy,and the nectotic area of flap of the rats in HPMSCs transfected with leptin group and HPMSCs group were less than that in blank control group.On the 7th day after administration, the survival rates of flap of the rats in HPMSCs transfected with leptin group and HPMSCs group were higher than that in blank control group(P<0.05).The survival rate of flap of the rats in HPMSCs transfected with leptin group was higher than that in blank control group(P<0.05). The HE staining results showed that on the 14th day after administration, there were the most abundant granulation tissues, new capillaries and fibroblasts and there were less inflammatory cell infiltration around the capillaries; the collagen arranged irregularly in HPMSCs transfected with leptin group.On the 14th day after administration, the immunohistochemical staining results showed that there were a lot of yellow particals (positive expression) in HPMSCs transfected with leptin group and HPMSCs group.A lot of leptin was shown in HPMSCs transfected with leptin group.Conclusion: In the role of local microenvironment, HPMSCs can differentiate into endothelial cells and promote the formation of new capillaries.HPMSCs and leptin have synergistic effects in the process of wound healing, and the HPMSCs with gene-modified or not could significantly increase the survival rate of flap after radiotherapy, and the HPMSCs transfected with leptin have better effect.

Key words: combined radiation-wound injury, leptin, wound healing, human placenta-derived mesenchymal stem cells

中图分类号:

Q813

费倩怡, 刘书会, 朱振威, 邹永巍, 郑明, 刘春丽, 刘志辉. 转染leptin的胎盘间充质干细胞对放射损伤复合创伤愈合的促进作用[J]. 吉林大学学报(医学版), 2016, 42(06): 1066-1070.

FEI Qianyi, LIU Shuhui, ZHU Zhenwei, ZOU Yongwei, ZHENG Ming, LIU Chunli, LIU Zhihui. Promotive effect of HPMSCs transfected with leptin on healing of combined radiation-wound injury[J]. Journal of Jilin University Medicine Edition, 2016, 42(06): 1066-1070.

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转染leptin的胎盘间充质干细胞对放射损伤复合创伤愈合的促进作用

Promotive effect of HPMSCs transfected with leptin on healing of combined radiation-wound injury

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