核因子-κB在神经干细胞增殖过程中的调控作用  

Regulation of NF-κB in neural stem cell proliferation

作　　者：汪泱[1] 刘玉霞[1] 娄远蕾[1] 郭菲[1] 涂伟[2] 邓志锋[2] 

机构地区：[1]南昌大学第一附属医院泌尿外科研究所,330006 [2]南昌大学第二附属医院神经外科

出　　处：《中华实验外科杂志》2010年第3期383-385,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30560156）;江西省自然科学基金资助项目（2007GZY1187）

摘　　要：目的观察核转录因子（NF）-κB信号通路的活化对体外培养的神经干细胞（NSCs）增殖的影响，探讨NF-κB在NSCs增殖过程中的调控作用。方法采用成球悬浮法分离培养小鼠神经干细胞（mNSCs）并鉴定NSCs标志蛋白nestin；将生长状态良好的第3代mNSCs分为空白对照组、肿瘤坏死因子-α（TNF-α）组、PDTC组，分别采用常规NSCs培养液、含10μg／LTNF-α的NSCs培养液、含0．1mmol／LPDTC的NSCs培养液培养30min、4h和72h；采用免疫荧光细胞化学方法及蛋白质免疫印迹法（Westernblot）检测NSCs的NF-κB活化状况；采用免疫荧光细胞化学方法检测增殖指标CyclinD1和Ki-67的表达，观察NSCs的增殖。结果培养的mNSCs表达nestin：免疫荧光细胞化学方法及Westernblot结果显示，TNF-α可明显使p65活化由胞质转位至胞核；TNF-α组和空白对照组，CyclinD1阳性细胞率分别为（68．6±4．2）％和（38．8±3．7）％，明显高于PDTC组的（24．2±2．8）％（P〈0．05）；Ki-67阳性细胞率分别为（48．2±3．6）％和（35．2±4．7）％，明显高于PDTC组的（18．7±1．8）％（P〈0．05）。结论NF-κB的激活或抑制可促进或抑制NSCs的增殖，NF-κB在NSCs的增殖过程中可能起着重要的调控作用。Objective To explore the role of nuclear factor-κB （NF-κB） in neural stem cells （NSCs） proliferation. Methods Neurospheres got from primary culture and subculture and identified by nestin, were used in the following studies. NSCs were divided randomly into 3 groups：contrel group, TNF-α group and PDTC group. In control group, NSCs were cultured in NSCs media routinely. In TNF-α group, the cells were cultured in NSCs media containing 10 μg/L TNF-α. NSCs in PDTC group were treated with NSCs media containing 0.1 mmol/L PDTC. The NSCs in each group were sampled at 0.5,4,72 h post-stimulation, respectively. The activity of NF-κB was detected by immunofluorescence staining and Western blotting. lmmunofluorescence staining was also employed to detect the expression of Cyelin D1 and Ki-67, the wellcharacterized proliferation markers. Results Immunofluorescence staining of nestin indicated that most cells in the neurospberes expressed nestin positively. Immunofluorescence staining and Western blotting results showed that NF-κB/p65 subunit was transloeated into the nuclear, suggesting the NF-κB pathway was activated in TNF stimulated NSCs. The positive rate of Cyclin D1 in TNF-α group and control group was （68.6 ±4.2） % and （ 38.8 ±3.7 ） %, respectively, both higher than in PDTC group significantly E （ 24.2 ±2.8） % ,P 〈 0.01 ]. The positive rate of Ki-67 in TNF-α group and control group was （48.2 ±3.6 ）% and （ 35.2 ±4.7 ） %, respectively, both higher than in PDTC group significantly [ （ 18.7±1.8 ） %, P 〈0. 01 ]. Conclusion The activation of NF-κB might promote the proliferation of NSCs and vice versa. NF-κB might play an important role in NSCs proliferation.

关 键 词：核转录因子-ΚB 神经干细胞 肿瘤坏死因子 增殖 

分 类 号：R285.5[医药卫生—中药学]