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目的研究白细胞介素6(IL-6)及其受体对体外培养的大鼠骨髓源性神经干细胞增殖的影响及信号转导机制。方法采用密度梯度离心法分离骨髓基质细胞,应用特制的神经干细胞培养基诱导培养6d后收集贴壁细胞传代培养,用AlamarBlue摄入法检测不同浓度的IL-6及IL-6受体对大鼠骨髓源性神经干细胞增殖的影响,Westernblot方法检测不同剂量IL-6对干细胞中STAT3表达的影响。结果IL-6能够促进干细胞的增殖,随着药物剂量的增加,增殖作用明显增加,呈明显剂量依赖性,其中20、100、200ng/mL组均与对照组有显著差异(P<0.01),但2ng/mL与对照组、100ng/mL与200ng/mL组之间无显著差异(P>0.05),20ng/mL与100ng/mL组之间有差异(P<0.05)。IL-6受体可抑制干细胞的增殖,单独应用IL-6受体,100ng/mL、200ng/mL组与对照组均有显著差异(P<0.05)。而IL-6与IL-6受体联合应用可促进干细胞的增殖,但增殖作用比单独应用IL-6组差,两组相比有显著性差异(P<0.05)。随IL-6浓度的增加,干细胞中的STAT3表达增加,20、100、200ng/mL组与对照组相差显著(P<0.01)。结论100ng/mL组的IL-6对骨髓源性神经干细胞有较好的增殖促进作用;IL-6受体可抑制骨髓源性神经干细胞增殖;联合应用IL-6和IL-6受体增殖效果不如单独应用IL-6,提示骨髓源性神经干细胞表面自身具有IL-6受体;IL-6通过增加STAT3表达促进骨髓源性神经干细胞增殖。
Objective To investigate the effects of interleukin-6 (IL-6) and its receptor on the proliferation of rat bone marrow-derived neural stem cells cultured in vitro and its signal transduction mechanism. METHODS: Bone marrow stromal cells were isolated by density gradient centrifugation. Adherent cells were harvested after passage of cultured neural stem cells (NSCs) for 6 days. The adherent cells were subcultured and the concentrations of IL-6 and IL-6 receptors were measured by AlamarBlue in rats The effect of different doses of IL-6 on STAT3 expression in stem cells was detected by Western blot. Results IL-6 could promote the proliferation of stem cells. With the increase of drug dose, the proliferation of IL-6 significantly increased in a dose-dependent manner. The levels of IL-6 in 20,100,200 ng / mL group were significantly different from those in control group (P <0.01) However, there was no significant difference between 2ng / mL and control group, 100ng / mL and 200ng / mL group (P> 0.05), but there was difference between 20ng / mL and 100ng / mL group (P <0.05). The IL-6 receptor could inhibit the proliferation of stem cells. IL-6 receptor alone, 100ng / mL, 200ng / mL group were significantly different from the control group (P <0.05). The combination of IL-6 and IL-6 receptor could promote the proliferation of stem cells, but the proliferative effect was worse than that of IL-6 alone group. There was significant difference between the two groups (P <0.05). With the increase of IL-6 concentration, the expression of STAT3 in stem cells increased, while there was a significant difference between 20,100 and 200ng / mL groups (P <0.01). CONCLUSION: IL-6 in 100ng / mL group can promote the proliferation of bone marrow-derived neural stem cells; IL-6 receptor can inhibit the proliferation of bone marrow-derived neural stem cells; combined with IL-6 and IL-6 receptor proliferation IL-6 alone, suggesting that the surface of bone marrow-derived neural stem cells itself has IL-6 receptor; IL-6 promotes proliferation of bone marrow-derived neural stem cells by increasing STAT3 expression.