hPlGF-2基因修饰的骨髓间充质干细胞对皮肤创伤修复及血管形成的作用研究  

Eeffect of hPlGF-2 Gene Modified BMSCs on Skin Wound Repairand Angiogenesis

作　　者：陈金逸 陈宗存[1] 饶朗毓[1] 黄亚莲 符茂雄 CHEN Jin-yi;CHEN Zong-cun;RAO Lang-yu;HUANG Ya-lian;FU Mao-xiong(Department of Endocrinology,the Second Affiliated Hospital of Hainan Medical College,Haikou 570311,Hainan,China)

机构地区：[1]海南医学院第二附属医院内分泌科,海南海口570311

出　　处：《中国美容医学》2020年第10期106-111,共6页Chinese Journal of Aesthetic Medicine

基　　金：海南省卫生计生委科研基金项目(编号:18A200065)。

摘　　要：目的:构建胎盘生长因子hPlGF-2基因腺病毒载体,探讨hPlGF-2修饰的骨髓间充质干细胞(Bone marro w mesenchymal stem cells,BMSCs)对SD大鼠皮肤愈合的影响。方法:分离培养SD大鼠BMSCs细胞,PCR法扩增hPlGF-2基因DNA片段,并构建pAdTrack-hPlGF-2重组载体;重组腺病毒pAd/hPlGF-2感染BMSCs后,倒置显微镜观察EGFP表达情况,PCR与Western blot分别检测hPlGF-2 mRNA和蛋白的表达,MTT法检测BMSCs增殖;构建SD大鼠皮肤割伤模型,以创缘注射感染重组腺病毒pAd/hPlGF-2的BMSCs为实验组,以创缘注射生理盐水为对照组,HE染色观察创面病理形态变化,免疫组织化学法检测带EGFP(+)的BMSCs在创面的分布情况。结果:成功构建了腺病毒载体pAdTrack-hPlGF-2,在hPlGF-2基因修饰的BMSCs中检测到了hPlGF-2 mRNA与蛋白的表达,且感染重组腺病毒pAd/hPlGF-2对BMSCs的增殖能力无影响。在大鼠皮肤创面注射hPlGF-2基因修饰的BMSCs后,创面感染程度较轻,愈合速度较快,在创伤后7d和14d观察到EGFP标记的BMSCs出现在创面血管周围和血管内皮处,同时有FⅧ表达。在创伤后7d新生肉芽组织中,bFGF的表达明显增加。结论:hPlGF-2基因修饰的BMSCs能分化为成纤维细胞参与到皮肤创面修复中,并分化为血管内皮细胞参与血管的再生。Objective An adenoviral vector of placental growth factor hPlGF-2 gene was constructed,and the effect of hPlGF-2 modified bone marrow mesenchymal stem cells(BMSCs)on skin healing in SD rats was investigated.Methods Isolate and culture SD rat BMSCs cells,amplify the hPlGF-2 gene DNA fragment by PCR,and construct the pAdTrack-hPlGF-2 recombinant vector;After infecting BMSCs with recombinant adenovirus pAd/hPlGF-2,observe the expression of EGFP by an inverted microscope.PCR and Western blot were used to detect the expression of hPlGF-2 mRNA and protein.MTT method to detect BMSCs proliferation;A model of SD rat skin cut was established.BMSCs infected with recombinant adenovirus pAd/hPlGF-2 were injected into the wound margin as the experimental group,and saline was injected into the wound margin as the control group.HE staining was used to observe the pathological changes of the wound.The distribution of BMSCs with EGFP(+)on the wound was detected by immunohistochemistry.Results The adenoviral vector pAdTrack-hPlGF-2 was successfully constructed,hPlGF-2 mRNA and protein expression was detected in hPlGF-2 gene-modified BMSCs,and infection with recombinant adenovirus pAd/hPlGF-2 had no effect on the proliferation capacity of BMSCs.After injecting hPlGF-2 genemodified BMSCs into rat skin wounds,the wound infection was milder and healed faster.EGFP-labeled BMSCs were observed around the blood vessels and endothelial endothelium at 7 and 14 days after trauma.At the same time,FVIII is expressed.The expression of bFGF was significantly increased in the new granulation tissue at 7 days after wounding.Conclusion The hPlGF-2 gene-modified BMSCs can participate in the repair of skin wounds.It is speculated that they differentiate into vascular endothelial cells and participate in the regeneration of blood vessels.

关 键 词：骨髓间充质干细胞 腺病毒载体 重组质粒 胎盘生长因子 基因修饰 创伤愈合 

分 类 号：R622[医药卫生—整形外科]