中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
7期
1395-1398
,共4页
崔猛%戴斌%冯世庆%辛景义
崔猛%戴斌%馮世慶%辛景義
최맹%대빈%풍세경%신경의
细胞因子信号传导抑制蛋白1%神经干细胞%神经元%分化
細胞因子信號傳導抑製蛋白1%神經榦細胞%神經元%分化
세포인자신호전도억제단백1%신경간세포%신경원%분화
Suppressor of cytokine signaling 1%Neuronal stem cells%Neuron%Differentiation
目的 观察细胞因子信号传导抑制蛋白1(SOCS1)对C17.2神经干细胞(C17.2NSCs)分化的调控作用.方法 用含目的基因SOCS1的慢病毒体外感染C17.2NSCs,实验分3组:SOCS1组、空载体组、磷酸盐缓冲液(PBS)组.采用光镜观察3组细胞的形态变化;采用细胞免疫荧光染色、Western blot、逆转录-聚合酶链反应(RT-PCR)法观察3组细胞中巢蛋白(Nestin)、微管相关蛋白Ⅱ(MAP2)、β-微管相关蛋白Ⅲ(β-tubulinⅢ)、人髓鞘碱性蛋白(MBP)、胶质纤维酸性蛋白(GFAP)的表达.结果 SOCS1组中Nestin表达较其他两组减少,而MAP2表达较其他两组增加(Nestin:0.234±0.008、0.925±0.040、1.000±0.039; MAP2:2.598±0.067、0.890±0.038、1.000±0.032,P ＜0.01);SOCS1组中部分C17.2NSCs的形态向神经元演变且该部分细胞均表达β-tubulinⅢ;SOCS1组中β-tubulinⅢ阳性染色细胞数所占比例高于其他两组(SOCS1组:10.05％,空载体组:0.71％,PBS组:0.65％,P＜0.01);SOCS1组中β-tubulinⅢ蛋白表达量较其他两组增加(3-tubulinⅢ:3.028±0.121、0.978±0.254、1.000±0.042,P＜0.01).结论 SOCS1能够调控C17.2NSCs向神经元分化.
目的 觀察細胞因子信號傳導抑製蛋白1(SOCS1)對C17.2神經榦細胞(C17.2NSCs)分化的調控作用.方法 用含目的基因SOCS1的慢病毒體外感染C17.2NSCs,實驗分3組:SOCS1組、空載體組、燐痠鹽緩遲液(PBS)組.採用光鏡觀察3組細胞的形態變化;採用細胞免疫熒光染色、Western blot、逆轉錄-聚閤酶鏈反應(RT-PCR)法觀察3組細胞中巢蛋白(Nestin)、微管相關蛋白Ⅱ(MAP2)、β-微管相關蛋白Ⅲ(β-tubulinⅢ)、人髓鞘堿性蛋白(MBP)、膠質纖維痠性蛋白(GFAP)的錶達.結果 SOCS1組中Nestin錶達較其他兩組減少,而MAP2錶達較其他兩組增加(Nestin:0.234±0.008、0.925±0.040、1.000±0.039; MAP2:2.598±0.067、0.890±0.038、1.000±0.032,P ＜0.01);SOCS1組中部分C17.2NSCs的形態嚮神經元縯變且該部分細胞均錶達β-tubulinⅢ;SOCS1組中β-tubulinⅢ暘性染色細胞數所佔比例高于其他兩組(SOCS1組:10.05％,空載體組:0.71％,PBS組:0.65％,P＜0.01);SOCS1組中β-tubulinⅢ蛋白錶達量較其他兩組增加(3-tubulinⅢ:3.028±0.121、0.978±0.254、1.000±0.042,P＜0.01).結論 SOCS1能夠調控C17.2NSCs嚮神經元分化.
목적 관찰세포인자신호전도억제단백1(SOCS1)대C17.2신경간세포(C17.2NSCs)분화적조공작용.방법 용함목적기인SOCS1적만병독체외감염C17.2NSCs,실험분3조:SOCS1조、공재체조、린산염완충액(PBS)조.채용광경관찰3조세포적형태변화;채용세포면역형광염색、Western blot、역전록-취합매련반응(RT-PCR)법관찰3조세포중소단백(Nestin)、미관상관단백Ⅱ(MAP2)、β-미관상관단백Ⅲ(β-tubulinⅢ)、인수초감성단백(MBP)、효질섬유산성단백(GFAP)적표체.결과 SOCS1조중Nestin표체교기타량조감소,이MAP2표체교기타량조증가(Nestin:0.234±0.008、0.925±0.040、1.000±0.039; MAP2:2.598±0.067、0.890±0.038、1.000±0.032,P ＜0.01);SOCS1조중부분C17.2NSCs적형태향신경원연변차해부분세포균표체β-tubulinⅢ;SOCS1조중β-tubulinⅢ양성염색세포수소점비례고우기타량조(SOCS1조:10.05％,공재체조:0.71％,PBS조:0.65％,P＜0.01);SOCS1조중β-tubulinⅢ단백표체량교기타량조증가(3-tubulinⅢ:3.028±0.121、0.978±0.254、1.000±0.042,P＜0.01).결론 SOCS1능구조공C17.2NSCs향신경원분화.
Objective To investigate the role of suppressor of cytokine signaling 1 (SOCS1) in regulating the differentiation of C17.2 neuronal stem cells (NSCs).Methods C17.2 NSCs were transfected by Lentiviral vector which included objective gene-SOCSl.The experiment was individed into LV-SOCS1 group,LV group and PBS group.The morphological change of C17.2NSCs after nransfection was observed by light microscopy.The expression of Nestin,microtubule-associated proteins 2 (MAP2),β-tubulinⅢ,myelin basic protein (MBP) and glial fibrillary acidic protein (GFAP) in C17.2 NSCs was detected by immunocytochemistry,Western blotting and reverse transcription-polymerase chain reaction (RT-PCR).The protein expression of β-tubulinⅢ in C17.2 NSCs was examined by immunocytochemistry and Western blotting.Results An decreased expression of Nestin and an increased expression of MAP2 in LV-SOCS1 group was found as compared with LV group and PBS group after transfection (P ＜ 0.01).Some C17.2 NSCs in LV-SOCS1 group underwent prominent neuronal morphological changes after transfection and almost all the cells with morphological changes were β-tubulin Ⅲ-positive,and the other two groups had less cells with morphological changes and β-tubulin Ⅲ-positive (LV-SOCS1 group:10.05％,LV group:0.71％,PBS group:0.65％) (P＜0.01).The β-tubulin lⅢ expression in LV-SOCS1 group was significantly higher than in other two groups (P ＜0.01).Conclusion SOCS1 may play a role in promoting the neuronal differentiation of C17.2 NSCs.
