肝细胞生长因子基因重组腺病毒修饰间充质干细胞修复急性放射性损伤大鼠的创面  被引量:3

Effects of adenovirus hepatocyte growth factor modified bone marrow mesenchymal stem cells on radioactive skin wound healing

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作  者:哈小琴[1] 张俊[1] 邓芝芸[1] 董菊子[1] 彭俊华[1] 赵勇[1] 张媛媛[1] 

机构地区:[1]解放军兰州军区兰州总医院检验科,甘肃省干细胞与基因药物重点实验室,甘肃省兰州市730050

出  处:《中国组织工程研究》2012年第49期9226-9231,共6页Chinese Journal of Tissue Engineering Research

基  金:全军"十一五"面上项目(06MB097);全军"十二五"面上项目(CWS11C229);兰州军区后勤科研计划项目(CLZ11J09)~~

摘  要:背景:基因药物与干细胞联合应用促进难治性大面积创面修复。目的:观察携带肝细胞生长因子基因的重组腺病毒(adenovirus hepatocyte growth factor,Ad-HGF)修饰的间充质干细胞对大鼠皮肤放射性损伤创面促愈合的作用。方法:体外分离、培养雄性Wistar大鼠骨髓间充质干细胞,转染Ad-HGF后备用。40只雌性Wistar大鼠用X射线局部照射制备急性放射性皮肤损伤模型。并将模型大鼠随机分为单纯间充质干细胞治疗组、单纯Ad-HGF治疗组、Ad-HGF修饰的间充质干细胞治疗组、溶剂对照组4组,制备模型即刻各组将相应细胞悬液、病毒悬液及溶剂行创面边缘多点注射。结果与结论:苏木精-伊红染色结果显示,伤后21d Ad-HGF修饰的间充质干细胞治疗组再生表皮明显薄于溶剂对照组,并可见丰富的新生小血管,且表皮较单纯间充质干细胞治疗组和单纯Ad-HGF治疗组更平整,新生小血管更丰富;免疫组织化学结果表明,Ad-HGF修饰的间充质干细胞治疗组创面中肝细胞生长因子表达均强于其他组;羟脯氨酸含量Ad-HGF修饰的间充质干细胞治疗组在7d后显著低于其他组(P<0.05);原位杂交结果显示,Ad-HGF修饰的间充质干细胞治疗组修复的皮肤组织中性别决定基因sry表达强于单纯间充质干细胞治疗组。结果提示,Ad-HGF修饰的间充质干细胞对皮肤放射性损伤创面有显著的促愈合修复作用,且优于二者单独应用。BACKGROUND: Combined application of gene therapy and stem cells can promote the healing of refractory large scale wound. OBJECTIVE: To observe the effect of adenovirus hepatocyte growth factor (Ad-HGF) modified bone marrow mesenchymal stem cells (BMSCs) on radioactive skin wound healing. METHODS: The BMSCs were in vitro isolated and cultured from male Wistar rats and transfected with Ad-HGF. Forty female Wistar rats were irradiated using X-ray therapy machine to establish acute skin radiation damage model. The models were randomly divided into four groups: BMSCs treatment group, Ad-HGF treatment group, Ad-HGF modified BMSCs treatment group and control group. After irradiating, the suspension of BMSCs, Ad-HGF and Ad-HGF modified BMSCs suspension were injected directly into the radioactive wounds. RESULTS AND CONCLUSION: Hematoxylin-eosin staining results showed that at 21 hours after injury, the epidermis in Ad-HGF modified BMSCs treatment group was significantly thinner than that in the control group, and the epidermis in the Ad-HGF modified BMSCs treatment group was more regular than that in the BMSCs treatment group and Ad-HGF treatment group, many small vessels were observed in the derma of radioactive wounds. Immunohistochemistry results showed that the expression of hepatocyte growth factor in Ad-HGF modified BMSCs treatment group was stronger than that in the other three groups; the content of hydroxyproline i n the Ad-HGF modified BMSCs treatment group was significantly lower than that in the other three groups after 7 days (P 〈 0.05); in situ hybridization showed that the expression of sex-determining sry gene in Ad-HGF modified BMSCs treatment group was stronger than that in the BMSCs treatment group. It demonstrates that Ad-HGF modified BMSCs can promote the hea ng of rad oact ve sk n wound, and the effect is better than simple application of the suspension.

关 键 词:间充质干细胞 肝细胞生长因子基因 重组腺病毒 放射损伤 修复 

分 类 号:R394.2[医药卫生—医学遗传学]

 

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