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

发布时间：2018-05-20 02:02

本文选题：骨髓 + 间质干细胞移植　；参考：《中国组织工程研究》2016年28期

【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(P0.05);(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(P0.05),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(P0.05);(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。
[Abstract]:Background: basic fibroblast growth factor (bFGF) has been shown to promote growth, proliferation, differentiation and functional expression of most cells derived from mesoderm and neuroectoderm. Aim: to investigate the effects of basic fibroblast growth factor gene modified bone marrow mesenchymal stem cell transplantation on renal function in rats with acute renal injury. Methods: the basic fibroblast growth factor gene was introduced into bone marrow mesenchymal stem cells according to adenovirus vector, and the expression of basic fibroblast growth factor in transfected cells was identified by RT-PCR method. The rat model of acute renal injury was made by clipping bilateral renal artery. The model was randomly divided into three groups: the negative transfection group was injected with untransfected bone marrow mesenchymal stem cell suspension via tail vein. Bone marrow mesenchymal stem cell suspension was injected via tail vein in basic fibroblast growth factor gene transfection group and DMEM culture medium was injected through tail vein in model group. Four weeks after transplantation, serum creatinine and urea nitrogen levels were detected. The expression of connective tissue growth factor and transforming growth factor 尾 1 in renal tissue were detected by Western blot method. The histopathology of renal tissue was observed by hematoxylin eosin staining. Results and conclusion: BGF gene was successfully expressed in bone marrow mesenchymal stem cells (BMSCs). Compared with the model group, the serum creatinine and urea nitrogen levels in the negative transfection group were significantly lower than those in the control group. The levels of serum creatinine and urea nitrogen in basic fibroblast growth factor gene transfection group were further decreased, and there was significant difference between each group and model group. The expression of connective tissue growth factor and transforming growth factor 尾 1 in renal tissue of negative transfection group and basic fibroblast growth factor gene transfection group was significantly decreased, while that of negative transfection group and basic fibroblast growth factor gene transfection group was significantly decreased. There was no significant difference in renal necrosis in negative transfection group. The inflammatory response was alleviated, and the renal tubular contours returned to normal in the basic fibroblast growth factor transfection group. The transplantation of bone marrow mesenchymal stem cells modified with basic fibroblast growth factor gene was more effective in renal repair of acute renal injury rats.
【作者单位】：济宁医学院解剖教研室;河北大学附属医院检验科;
【分类号】：R692

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