核因子-κB在黄芩甙诱导大鼠骨髓基质细胞分化为神经细胞中的作用

作者	单位
杨琴	重庆医科大学附属第一医院神经科
贾延劼	重庆医科大学附属第一医院神经科
杨军	重庆医科大学附属第一医院神经科
董为伟	重庆医科大学附属第一医院神经科
杨于嘉	中南大学湘雅医院儿科

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中文摘要:

目的研究核因子 κB(NF- κB)在黄芩甙诱导骨髓基质细胞分化为神经细胞中的作用。方法在无血清条件下 ,以中药单体黄芩甙作为主要诱导剂 ,诱导大鼠骨髓基质细胞分化为神经细胞 ;同时以无血清培养基为对照组。采用免疫荧光细胞化学染色检测神经细胞标记蛋白表达和NF-κB亚基P6 5核移位情况 ;蛋白质印迹 (Westernblot)检测细胞中NF-κB抑制蛋白 (IκBα)表达变化 ;原位末端标记 (TUNEL法 )染色评价细胞凋亡率。结果黄芩甙诱导后细胞形成较典型的神经细胞形态 ,同时表达多种神经细胞标记蛋白。对照组无类似情况 ,但出现P6 5由胞浆向胞核移位 ,细胞内IκBα表达水平显著降低 ,细胞凋亡率为 (2 8.2±6.1) % ;黄芩甙诱导后P6 5信号主要仍在胞浆中表达 ,细胞内IκBα表达水平降低程度较少 ,细胞凋亡率〔(12 .2± 2.8) %〕 ,也显著低于对照组 (P <0. 0 1)。结论黄芩甙可抑制NF κB的活化 ,在骨髓基质细胞诱导分化为神经细胞的过程中可能起到一定的作用

英文摘要:

explore the effect of nuclear factor-κB (NK-κB) on differentiation of bone marrow stromal cells (MSC) into neurons induced by baicalin. MethodsUsing baicalin as main inducer, differentiation of rats’ MSC was induced into NC in serum-free medium, at the same time, untreated cells were cultured in serum-free medium as the control group. The expression of NC special marker protein and the presence of NK-κB subunits RelA (P65) translocated into nucleus were determined by immunofluorescent cytochemical stain, the change of expression of NK-κB inhibitory protein (IκBα) were determined by Western blot, and the apoptotic index was assessed by terminal deoxynucleotidyl transferase-mediated d-UTP-biotin nick end labeling (TUNEL) assay. ResultsMSC displayed typical shape of NC and expressed several NC marker protein after induced by baicalin, while those changes were not revealed in the control group. but showed translocation of P65 from cytoplasm to nucleus, and the markedly reduction of IκBα expression, the apoptotic index being 28.2±6.1%. After induced by baicalin, the P65 still revealed mainly in cytoplasm with the level of IκBα expression decreased lesser and the number of apoptosis cell (12.2±2.8%) lower than those in the control group, P<0.01. ConclusionBaicalin could inhibit NK-κB activation, and it has certain effect in inducing the differentiation of MSC into NC.

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