碱性成纤维细胞生长因子基因修饰骨髓间充质干细胞移植修复急性肾损伤  

Basic fibroblast growth factor-transfected bone marrow mesenchymal stem cell transplantation for acute kidney injury

作　　者：崔利德[1] 李铁民[2] 

机构地区：[1]济宁医学院解剖教研室,山东省济宁市272029 [2]河北大学附属医院检验科,河北省保定市071000

出　　处：《中国组织工程研究》2016年第28期4169-4175,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:研究表明,碱性成纤维细胞生长因子对源于中胚层和神经外胚层的大部分细胞均有促进生长、增殖、分化及功能表达的作用。目的:探讨碱性成纤维细胞生长因子基因修饰骨髓间充质干细胞移植对急性肾损伤大鼠肾功能的影响。方法:依据腺病毒载体将碱性成纤维细胞生长因子基因导入骨髓间充质干细胞,采用RT-PCR法鉴定转染细胞中碱性成纤维细胞生长因子的表达。采用动脉夹夹闭双侧肾蒂法制作急性肾损伤大鼠模型,建模后随机分成3组(n=20):阴性转染组经尾静脉注射未经转染的骨髓间充质干细胞悬液,碱性成纤维细胞生长因子基因转染组经尾静脉注射转染成功的骨髓间充质干细胞悬液,模型组经尾静脉注射DMEM培养液。移植后4周,检测血清肌酐及尿素氮水平,Western blot法检测肾组织中结缔组织生长因子、转化生长因子β1的表达,苏木精-伊红染色观察肾组织病理形态。结果与结论:(1)碱性成纤维细胞生长因子基因成功表达于骨髓间充质干细胞中;(2)与模型组比较,阴性转染组大鼠血清肌酐及尿素氮水平明显降低,碱性成纤维细胞生长因子基因转染组血清肌酐及尿素氮水平进一步降低,各组间比较差异有显著性意义(P<0.05);(3)与模型组比较,阴性转染组及碱性成纤维细胞生长因子基因转染组肾组织结缔组织生长因子及转化生长因子β1表达明显减弱(P<0.05),阴性转染组及碱性成纤维细胞生长因子基因转染组间差异无显著性意义(P>0.05);(4)阴性转染组肾组织坏死、炎性反应减轻,碱性成纤维细胞生长因子转染组肾小管轮廓恢复正常,未出现明显坏死细胞;(5)结果表明,碱性成纤维细胞生长因子基因修饰的骨髓间充质干细胞移植对急性肾损伤大鼠肾脏的修复作用更明显。BACKGROUND：It has been confirmed that basic fibroblast growth factor （bFGF） can promote the growth, proliferation, differentiation and functional expression of most cels derived from neuroderm and mesoderm. OBJECTIVE：To investigate the effect of bFGF-transfected bone marrow mesenchymal stem cel transplantation in rats with acute kidney injury. METHODS： bFGF genes were transfected into bone marrow mesenchymal stem cels via an adenovirus vector, and then expression of bFGF in transfected cels was identified using RT-PCR technology. Rat models of acute kidney injury were prepared by clipping bilateral renal pedicles, and then randomized into three groups （n=20）： rats were given injection of bone marrow mesenchymal stem cel suspensionsvia tail vein as negative transfected group, those given injection of bFGF-transfected bone marrow mesenchymal stem cel suspensionsvia tail vein as bFGF-transfected group, and the others given injection of DMEMvia tail vein as model group. Four weeks later, levels of serum creatinine and urea nitrogen were detected, expressions of connective tissue growth factor and growth factor in renal tissues were detected by Western blot assay, and morphology of renal tissues was observed using hematoxylin-eosin staining. RESULTS AND CONCLUSION：bFGF genes were successfuly transfected into bone marrow mesenchymal stem cels. Compared with the model group, the levels of serum creatinine and urea nitrogen were significantly reduced in bFGF-transfected and negative transfected groups, especialy in the bFGF-transfected group （P 〈 0.05）, while expressions of connective tissue growth factor and transforming growth factor in renal tissues in bFGF-transfected and negative transfected groups were significantly weakened in these two groups （P 〈 0.05）, but there were no significant differences between the bFGF-transfected group and negative transfected group （P 〉 0.05）. Besides, renal tissues necrosis and inflammatory reactions were mitigated in the negative transfected group

关 键 词：骨髓 间质干细胞移植 成纤维细胞生长因子2 肾疾病 组织工程 干细胞 移植 碱性成纤维细胞生长因子 骨髓间充质干细胞 基因修饰 大鼠 急性肾损伤 肾功能 

分 类 号：R394.2[医药卫生—医学遗传学]