摘要
目的研究携带人肝细胞生长因子基因的重组腺病毒（Ad—HGF）修饰骨髓间充质干细胞（MSCs）对大鼠脂肪颗粒移植存活率的影响。方法用携带绿色荧光蛋白的重组腺病毒（Ad-GFP）、Ad—HGF转染雄性Wistar大鼠MSCs，测定转染效率与感染上清中肝细胞生长因子（HGF）的表达。用雌性Wistar大鼠150只，随机分为5组：空白对照组（A组）、Ad—HGF组（B组）、MSCs组（C组）、Ad—GFP感染MSCs组（D组）和Ad—HGF感染MSCs组（E组），每组30只。各组均匀振动5分钟，将颗粒脂肪移植于自体背部皮下；移植后3、5、7、14、28、60d，每组各取5只大鼠脱颈处死，取出移植物；测体积、HE、Masson染色观察病理改变，免疫组织化学法测定组织中HGF表达和血管生成。结果积分吸光度＝100时，Ad—GFP转染MSCs效率可达89．6％；移植后3、5、7、14d，E组移植物中HGF的表达高于其他各组（P〈0．05）；移植后5、7、14、28、60d，E组血管数量也多于其他各组（P〈0．05）；28d和60d时，E组脂肪颗粒体积保持率明显好于其他4组（P〈0．05）。结论携带HGF基因的MSCs在移植物中能够持续表达HGF，有效地减少脂肪颗粒移植后的吸收，从而提高移植颗粒脂肪的存活率。
Objective To investigate the effect of mesenchymal stem cells （MSCs） transfected with hepatic growth factor （HGF） gene on the survival volume of free grain fat grafts. Methods MSCs of male Wistar rats were transfected with Ad-GFP or Ad-HGF. The transfection infectivity of Ad-GFP to MSCs, the expression of HGF were measured using ELISA assay. 150 rats were randomly divided into 5 groups： control group （group A）, MSCs group （group B）, Ad-HGF group （group C）, MSCs transplanted with Ad-GFP group （group D）, and MSCs transplanted with Ad-HGF （MSCs- HGF） group （group E）. Then, the same volume of frain fat graft, mixed with DMEM-LG andMSCs, Ad-HGF, MSCs-GFP, MSCs-HGF respectively, were transplanted to rats back, but control group were only mixed with DMEM-LG. Fat graft was obtained on days 3, 5, 7, 14, 28, and 60 after im- plantation. The volume of fat graft was measured by messcylinder, and the expression of HGF and CD34 in transplanted fat tissue were detected by immunohistochemistry. Results The transfection infectivity of Ad-GFP to MSCs was 89.6 % at 100 MOI, the expression of HGF in MSCs culture medi- um reached to the level after being transfected with Ad-HGF for 48 h. Compared with other 4 groups, at days 3, 5, 7, and 14 post-transplantation, the expression of HGF in E group transplanted fat of group E had statistics significance （P〈0.05）. The persentage of survival volume of fat graft in group E was significantly higher than that of other group （P〈0.05） at days 28, and 60 post-transplanta- tion. Conclusions MSCs transplanted with Ad-HGF could secrete HGF and increase the survival volume of fat grafts.
出处
《中华医学美学美容杂志》
2011年第6期433-436,共4页
Chinese Journal of Medical Aesthetics and Cosmetology
基金
全军“十一五”医学科学技术研究面上项目（编号：06MA079）
甘肃省自然科学基金资助项目（编号：3ZS061-A25-099）
关键词
脂肪颗粒
自体
骨髓间充质干细胞
肝细胞生长因子
转染
基因治疗
Fat grain grafts, Autologous
Mesenchymal stem cells
Hepatic growth factor, Transfection
Gene therapy
