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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
Objective To investigate the role of nuclear factor-κB (NF-κB) in the differentiation of bone marrow stromal cells into neural cells induced by Astragalus membranaceus. METHODS: Serum-free medium was used as a control group to induce rat bone marrow stromal cells to differentiate into neural cells under the condition of serum-free conditions, with Chinese herbal medicine Huangqi as the main inducer. Immunofluorescence staining was used to detect the expression of neuronal marker protein and nuclear translocation of NF-κB subunit P6 5; Western blot was used to detect the expression of NF-κB inhibitory protein (IκBα) in cells; in situ end labeling (TUNEL) Staining was used to evaluate the rate of apoptosis. Results After induced by Astragalus mongholicus, the cells formed more typical neuronal morphology and expressed a variety of neuronal marker proteins. There was no similar situation in the control group, but there was a migration of P6 5 from the cytoplasm to the nucleus. The expression level of IκBα in the cells was significantly decreased, and the apoptosis rate was (2 8.2 ± 6.1)%; the signal of P6 5 remained mainly after induction of Astragalus membranaceus. In cytoplasm, the expression of intracellular IκBα was decreased, and the rate of apoptosis was (12.2±2.8)%, which was also significantly lower than that of the control group (P<0.01). Conclusion Astragalus can inhibit the activation of NF-κB and may play a role in the differentiation of bone marrow stromal cells into neural cells.