CXCR4转染的骨髓间充质干细胞靶向治疗急性肝衰竭  

Genetically modified mesenchymal stem cells overexpressing CXCR4 show increased colonization ability and confer better liver regeneration in mice

作　　者：马虎成[1] 施晓雷[1] 任昊桢[1] 袁献温[1] 丁义涛[1] 

机构地区：[1]南京大学医学院附属鼓楼医院肝胆外科,江苏省南京市210008

出　　处：《世界华人消化杂志》2014年第16期2229-2236,共8页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.81170418;江苏省自然科学基金资助项目;No.BK20131084~~

摘　　要：目的:提高骨髓间充质干细胞(mesenchymal stem cell,MSC)移植治疗急性肝衰竭时在肝脏的定植率及疗效.方法:采用慢病毒转染使MSC过表达CXCR4基因,RT-PCR和细胞流式检测CXCR4表达.ELISA实验检测肝衰竭裸鼠肝脏MSC向基质衍生因子-1α(stromal cell-derived factor-1α,SDF-1α)水平.迁移实验检测转染的SDF-1α趋化能力.20%(V/V)四氯化碳(CCl4)橄榄油溶液8μL/g腹腔注射诱导裸鼠急性肝衰竭模型,24 h后尾静脉注射CXCR4转染的间充质干细胞(CXCR4-MSC)和未转染的间充质干细胞(Null-MSC).于注射后各时间点活体示踪MSC分布情况、裸鼠肝功能、生存率、组织病理学改变和肝细胞增殖情况.结果:CXCR4-MSC高表达CXCR4基因和CXCR4蛋白,肝衰竭裸鼠肝脏SDF-1α水平升高,与正常裸鼠相比有统计学差异.体外迁移实验证实CXCR4-MSC比Null-MSC具有向SDF-1α更好的迁移能力.体内活体成像显示移植后5 d CXCR4-MSC主要定植在肝脏,而Null-MSC在肝脏和脾脏都有定植.高定植率使得转染组裸鼠比未转染组裸鼠肝细胞受损更轻且具有更长的生存期,同时转染组裸鼠肝细胞增殖明显多于未转染组,两者有统计学差异.我们的实验证实转染CXCR4基因的骨髓间充质干细胞能够更好地促进肝再生.结论:基因修饰使骨髓间充质干细胞过表达CXCR4基因能够促进干细胞在肝脏的定植,改善肝功能,提高肝细胞的再生和裸鼠的生存率,为干细胞移植治疗肝功能衰竭提供了更广阔的应用途径.AIM： To explore whether mesenchymal stem cells（MSCs） overexpressing CXCR4 show increased colonization ability and confer better liver regeneration in mice.METHODS： MSCs were modified with CXCR4gene（CXCR4-MSCs） or not（Null-MSCs） through lentiviral transduction. The characteristics of CXCR4-MSCs and Null-MSCs were determined by RT-PCR and flow cytometry. CXCR4-MSCs and Null-MSCs were infused intravenously 24h after administration of CCl4 in nude mice. The concentration of SDF-1α in the damaged liver was detected by ELISA. Transwell migration assays were carried out to evaluate the migration ability of MSCs toward SDF-1α. The distribution of the stem cells, their survival rates, liver function, histopathology and hepatocyte regeneration were analyzed. RESULTS： Transfected MSCs overexpressed CXCR4 at both gene and protein levels. In vitro, CXCR4-MSCs showed better migration capability toward SDF-1α. In vivo imaging showed that CXCR4-MSCs migrated to the liver in greater numbers than Null-MSCs 5 d after intravenous infusion in mice with acute liver failure（ALF）. Higher colonization led to a longer lifetime and better liver function. Immunohistochemistry analysis of Ki-67 showed increased cell proliferation in the damaged liver of CXCR4-MSCtreated animals. CONCLUSION： Genetically modified MSCs expressing CXCR4 show greater colonization ability and confer better functional recovery of the damaged liver.

关 键 词：急性肝衰竭 细胞移植 间充质干细胞 趋化因子受体4 基因修饰 

分 类 号：R575.3[医药卫生—消化系统]