摘要
目的探讨超声辐照下携基质衍生因子1(SDF-1α)基因的胞膜核膜双靶向阳离子微泡载体系统促进内源性干细胞归巢的机制。方法构建双靶向阳离子微泡载体系统及携SDF-1α阳离子微泡载体系统,分别于体外转染人脐静脉血管内皮细胞(HUVEC)并检测SDF-1α质粒入胞率、入核率及SDF-1α基因表达;进而构建心梗兔模型,造模成功后随机分为三组进行SDF-1α基因转染:A组:携SDF-1α双靶向阳离子微泡载体组;B组:携SDF-1α阳离子微泡载体组;C组:对照组。分别于基因转染后3天和4周后检测SDF-1α基因表达、干细胞标志物和血管新生效应。结果体外实验中,超声辐照下,与携SDF-1α阳离子微泡载体系统比较,携SDF-1α双靶向阳离子微泡载体系统SDF-1α质粒入胞率与其无显著差异(P>0.05),但入核率和SDF-1α蛋白表达显著增高(P<0.01);体内实验显示,与阳离子微泡载体组及对照组比较,双靶向阳离子微泡载体组家兔心肌组织中SDF-1α蛋白显著增高、干细胞标志物阳性染色显著增强、毛细血管密度显著增加(P<0.01)。结论超声辐照下携SDF-1α的胞膜核膜双靶向阳离子微泡载体系统能有效提高SDF-1α基因载体转染效率,表达足量的SDF-1α蛋白并发挥其干细胞诱导剂作用,从而促进内源性干细胞归巢并促进局部血管新生。
Objective To evaluate the mechanisms of endogenous stem cell homing improvement by cytop1αsmic and nuclear membrane double targeted cationic microbubbles carrying SDF-1α gene combined with ultrasonic irradiation. Methods The double targeted cationic microbubbles carrier system and cationic microbubbles carrying SDF-1α gene were constructed. SDF-1α gene was transfected to the human umbilical vein endothelial cell in vitro and efficiency of SDF-1α gene transfection was detected including the SDF-1αp1αsmid cellu1αrimport rate, the SDF-1αp1αsmidnuclear import rate and gene expression of SDF-1α protein. Then the myocar- dial infarction rabbits models were built and divided randomly into A group, B group and C group. A group was treated with double targeted cationic microbubbles carrying SDF-1α group. B group was treated with cationicmicrobubbles carrying SDF-1αgroup. C group was control group. 3 days and 4 weeks after gene transfection, gene expression of SDF-1α gene, stem cell markers and angio- genesis effects were detected respectively. Results In vitro, under the ultrasonic irradiation, compared with the cation microbubbles carrying SDF-1α system, there was no difference in the SDF-1αp1αsmidcellu1αr import rate between the cation microbubbles carrying SDF-1α system and the double targeted cationic microbubbles carrying SDF-1α system, while the 1αtter had the higher the SDF- 1αp1αsmidnuclear import rate and gene expression of SDF-1α protein. In vivo, compared with thecationic microbubble carrying SDF- 1α group and control group, SDF-1α protein expression, stem cell marker positive staining and capil1αry density in rabbits＇ myocardi- um increased significantly in the double targeted cationic microbubble carrying SDF-1α group. Conclusion Cytop1αsmic and nuclear membrane double targeted cationic microbubbles carrying SDF-1α gene combined with ultrasonic irradiation can improve the SDF-1α gene transfection efficiency effectively, which induces the sufficient expression of SDF-1α protei
作者
冯闯丽
陈金玲
邓倾
梅丹娥
胡波
曹省
周青
FENG Chuangli CHEN Jinling DENG Qing MEI Dane HU Bo CAO Sheng ZHOU Qing(Department of Ultrasound, Renmin Hospital of Wuhan University, Wuhan 430060, Chin)
出处
《西部医学》
2017年第4期462-466,473,共6页
Medical Journal of West China
基金
国家自然科学基金(81101058
81471674
81501495)
