中华神经外科杂志
中華神經外科雜誌
중화신경외과잡지
Chinese Journal of Neurosurgery
2014年
2期
186-189
,共4页
龙乾发%汪平%王安生%韩蕊%汪凯%贾晨光%费舟%刘卫平
龍乾髮%汪平%王安生%韓蕊%汪凱%賈晨光%費舟%劉衛平
룡건발%왕평%왕안생%한예%왕개%가신광%비주%류위평
骨髓间充质干细胞%发状分裂相关增强子-1%分化
骨髓間充質榦細胞%髮狀分裂相關增彊子-1%分化
골수간충질간세포%발상분렬상관증강자-1%분화
Bone marrow mesenchymal stem cells%Hes1%Differentiation
目的 探索Hes1基因沉默对骨髓间充质干细胞(BMSCs)向GABA能神经元分化的影响.方法 体外分离纯化2～3周的SD大鼠BMSCs,通过RNAi技术沉默BMSCs中的Hes1基因,而后条件诱导BMSCs向GABA能神经元分化,最后通过免疫细胞化学和Western blot分析细胞分化趋向.结果 Hes1沉默的BMSCs经条件诱导后,免疫细胞化学结果示GAD67和NeuN阳性细胞数分别为(35.7±2.8)％和(72.4±3.2)％,均明显多于对照组(P＜0.05);Western blot结果示实验组GAD67蛋白条带灰度值与β-aetin的比值(GAD67/ β-actin)为(0.282 ±0.042),NeuN/β-actin为(0.728±0.039),同样高于对照组(P＜0.01).结论 Hes1基因沉默可促进BMSCs向GABA能神经元分化.
目的 探索Hes1基因沉默對骨髓間充質榦細胞(BMSCs)嚮GABA能神經元分化的影響.方法 體外分離純化2～3週的SD大鼠BMSCs,通過RNAi技術沉默BMSCs中的Hes1基因,而後條件誘導BMSCs嚮GABA能神經元分化,最後通過免疫細胞化學和Western blot分析細胞分化趨嚮.結果 Hes1沉默的BMSCs經條件誘導後,免疫細胞化學結果示GAD67和NeuN暘性細胞數分彆為(35.7±2.8)％和(72.4±3.2)％,均明顯多于對照組(P＜0.05);Western blot結果示實驗組GAD67蛋白條帶灰度值與β-aetin的比值(GAD67/ β-actin)為(0.282 ±0.042),NeuN/β-actin為(0.728±0.039),同樣高于對照組(P＜0.01).結論 Hes1基因沉默可促進BMSCs嚮GABA能神經元分化.
목적 탐색Hes1기인침묵대골수간충질간세포(BMSCs)향GABA능신경원분화적영향.방법 체외분리순화2～3주적SD대서BMSCs,통과RNAi기술침묵BMSCs중적Hes1기인,이후조건유도BMSCs향GABA능신경원분화,최후통과면역세포화학화Western blot분석세포분화추향.결과 Hes1침묵적BMSCs경조건유도후,면역세포화학결과시GAD67화NeuN양성세포수분별위(35.7±2.8)％화(72.4±3.2)％,균명현다우대조조(P＜0.05);Western blot결과시실험조GAD67단백조대회도치여β-aetin적비치(GAD67/ β-actin)위(0.282 ±0.042),NeuN/β-actin위(0.728±0.039),동양고우대조조(P＜0.01).결론 Hes1기인침묵가촉진BMSCs향GABA능신경원분화.
Objective To study the influence of Hes1 gene silencing on the GABAergic differentiation of bone marrow mesenchymal stem cells (BMSCs).Methods BMSCs were isolated from 2-3 weeks SD rats and purified in vitro,then characterized by flow cytometry.Mter that,Hes1 gene of BMSCs was inhibited by RNAi technology,then BMSCs (control group) and Hes1 gene silencing BMSCs (experimental group) were induced by conditioned medium (10 μmol of retinoic acid,1 mmol N5,O2'-dibutyryl adenosine 3':5'-cyclic monophosphate,2％ B27) respectively.Finally,immunocytochemistry and Western blot were employed to analyze the differentiation potential of the induced cells.Results Experimental group and control group were induced by conditioned medium for 3-7 days,immunocytochemistry results showed that the percentage of GAD67-and NeuNpositive cells were (35.7± 2.8)％ and (72.4 ± 3.2)％ in the experimental group,which were more than the control group significantly (P ＜ 0.05) ; Western blot results also showed that the protein gray value ratio of GAD67 or NeuN compared to the β-actin in the experiment was significantly higher than the control group through Bandscan software analysis (P ＜0.01).Conclusions Hes1 gene silencing can promote BMSCs to differentiate into GABAergic neurons.
