CAJ | 학술논문

目的:探讨胶质细胞源性神经营养因子(glial cell line-derived neurotrophic factor, GDNF)基因修饰的骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)向神经元样细胞的分化以及神经营养因子的表达.方法:用GDNF重组腺病毒载体和空病毒质粒(blank virus plasmid,BVP)分别感染大鼠BMSCs 2d(称作GDNF/BMSCs和BVP/BMSCs),用免疫荧光染色法检测细胞的神经元特异性标志物微管相关蛋白(microtubule-associated protein 2, MAP2)的阳性细胞数量,PCR 法检测细胞MAP2和GAP-43 mRNA的表达,ELISA法检测细胞上清液中GDNF和NGF的表达.结果:BVP/BMSCs组未见MAP2阳性细胞,不表达MAP2和GAP-43 mRNA,GDNF/BMSCs组MAP2阳性细胞率为(42.21±4.79)%,表达MAP2和GAP-43 mR-NA,其上清液中的GDNF和NGF蛋白含量高于空病毒感染组(P<0.05).结论:GDNF具有促进BMSCs向神经元样细胞分化的作用,其分化作用可能与GAP-43,GDNF和NGF的表达上调有关.
목적:탐토효질세포원성신경영양인자(glial cell line-derived neurotrophic factor, GDNF)기인수식적골수간충질간세포(bone marrow mesenchymal stem cells,BMSCs)향신경원양세포적분화이급신경영양인자적표체.방법:용GDNF중조선병독재체화공병독질립(blank virus plasmid,BVP)분별감염대서BMSCs 2d(칭작GDNF/BMSCs화BVP/BMSCs),용면역형광염색법검측세포적신경원특이성표지물미관상관단백(microtubule-associated protein 2, MAP2)적양성세포수량,PCR 법검측세포MAP2화GAP-43 mRNA적표체,ELISA법검측세포상청액중GDNF화NGF적표체.결과:BVP/BMSCs조미견MAP2양성세포,불표체MAP2화GAP-43 mRNA,GDNF/BMSCs조MAP2양성세포솔위(42.21±4.79)%,표체MAP2화GAP-43 mR-NA,기상청액중적GDNF화NGF단백함량고우공병독감염조(P<0.05).결론:GDNF구유촉진BMSCs향신경원양세포분화적작용,기분화작용가능여GAP-43,GDNF화NGF적표체상조유관.
Objective: To investigate the differentiation of GDNF gene-modified bone marrow mesenchymal stem cells into neuron-like cells and the expression of neurotrophic factors. Methods: GDNF recombinant aden-ovirus vector and blank virus plasmid (BVP) were used to infect rat BMSCs, to obtain GDNF-BMSCs and BVP-BMSCs. Immunofluorescence staining was used to observe the number of neuron specific marker,the mircotubule-associated protein 2 (MAP2)-positive cells.RT-PCR technique was used to examine the expression levels of MAP2 and GAP-43 mRNA in cell pellets. ELISA assay was employed to detect the expression levels of GDNF and NGF protein in supernatant. Results: In the BVP/BMSCs group, no MAP2-positive staining was observed and MAP2 and GAP-43 mRNA expression were nearly undetectable. In the GDNF/BMSCs group, MAP2-positive rate was (42.21±4.79)% and the mRNA of MAP2 and GAP-43 was observed.The expression of GDNF and NGF proteins was higher in the GDNF/BMSCs group than in the BVP/BMSCs group (P<0.05). Conclusion:GDNF has the effect of promoting the differentiation of BMSCs to neuron-like cells, and its effect may be correlated with the up-regu-lation of expression of GAP-43, GDNF and NGF.