异体移植工程细胞对大鼠烧伤创面愈合中皮肤附属器作用  

Effects of mesenchymal stem cells transfected with human hepatocyte growth factor gene on formating appendant organ to heal burn wounds

作　　者：赵曼[1] 他维玮[1] 哈小琴[2] 惠玲[2] 贾庆华[2] 王美亮[2] 刘国军[3] 

机构地区：[1]兰州大学第二临床医学院,2009级兰州市730030 [2]兰州军区兰州总医院医学实验中心,兰州市730050 [3]兰州大学公共卫生学院,2009级兰州市730030

出　　处：《中华实用诊断与治疗杂志》2011年第9期846-849,共4页Journal of Chinese Practical Diagnosis and Therapy

基　　金：全军"十一五"B类项目(06MB097)

摘　　要：目的以大鼠深Ⅱ度烧伤为模型,观察工程细胞即腺病毒介导肝细胞生长因子(hepatocyte growth factor,HGF)基因修饰的骨髓间充质干细胞(mesenchymal stem cells,MSCs)对烧伤创面愈合中皮肤附属器再生的作用。方法体外分离、培养雄性Wistar大鼠骨髓MSCs,转染携带HGF基因的重组腺病毒(100 pfu/cell)后备用。16只雌性Wistar大鼠复制深Ⅱ度烧伤模型,随机分为观察组与对照组各8只。在造模同时,观察组将携带HGF基因的重组腺病毒修饰的骨髓MSCs悬液,对照组将同体积磷酸盐缓冲液注射于创面真皮下。分别于细胞移植后第3,5,7,14,21天计算创口愈合率,第7,14,21天光镜下观察组织愈合及皮肤附属器情况,常规PCR法检测细胞移植后第21天创面组织中性别决定基因Sry片段。结果携带HGF基因的重组腺病毒可有效转染MSCs,修饰的MSCs移植后第7,14天,观察组创口收缩率与对照组比较,差异有统计学意义(P<0.01)。HE染色结果显示,第7天观察组表皮化优于对照组,第21天观察组再生表皮明显厚于对照组,并可见再生毛囊和皮脂腺。观察组雌性大鼠创面组织中扩增到Sry基因片段。结论 MSCs联合携带HGF基因的重组腺病毒异体移植可促进烧伤创面愈合修复过程中皮肤附属器的形成,且移植的细胞也参与了烧伤创面修复过程中附属器的形成。Objective To explore the effects of bone marrow-derived mesenchymal stem cells （MSCs） transfected with adenoviral vector mediated hepatocyte growth factor （Ad-HGF） on formating appendant organ to heal burn wounds in rats with skin burn（ Ⅱ ）. Methods MSCs from a male Wistar rat were cultured and transfected with Ad-HGF（100 pfu/cell）. Another 16 female rats were established models of partial thickness skin burn（ H ） and were randomly divided into observation group and control group with 8 rats in each group. In the process of modeling, the observation group carried Ad-HGF modified bone marrow MSCs suspension, and control group were injected with the same volume of phosphate buffer solution in the derma of burn wounds. On the3rd, 5th, 7th, 14th, and 21st day after burn, the wound healing rates were calculated and state of skin appendages were observed under the light microscope on the 7th, 14th and 21st day. The Sry sex-determining gene fragments were detected in wound tissue with PCR technique on the 21st day. Results Adenovirus carrying the HGF gene transfected effectively MSCs. On the 7th and 14th day after the modified MSCs was transplanted, there was significant difference in wound contraction rate between the observation group and control group（P〈0.01）. The histological examination showed that the range of re-epidermis was significantly larger on the 7th day after burn, and the thickness of epidermis was significantly thicker on the 21st day after burn in the observation group than that in control group. The hair follicle and sebacenous glands were found regenerated in observation group. The wound tissue in female rats transplanted the male rats＇ cells was amplified into the Sry gene. Conclusion Transplantation of MSCs transfected with Ad-HGF has positive effect on formating appendant organ to heal burn wounds, and transplanted cells are involved in the appendages formation of wound healing.

关 键 词：烧伤 间充质干细胞 肝细胞生长因子 愈合 皮肤附属器 

分 类 号：R644[医药卫生—外科学]