激活的星形胶质细胞分泌聚集素蛋白及其对海马神经干细胞向神经元分化的促进作用

发布时间：2018-07-13 17:14

【摘要】：目的探讨切割穹隆海马伞的侧海马提取液"激活"的星形胶质细胞分泌聚集素蛋白(CLU)及体外CLU诱导海马神经干细胞分化为神经元的情况。方法分别用切割穹隆海马伞的海马提取液、正常海马提取液、DMEM/F12培养基培养星形胶质细胞,制备切割12h、24h、48h组,正常组和对照组条件培养基;ELISA检测比较不同条件培养基中CLU蛋白的浓度;体外细胞培养采用CLU诱导海马神经干细胞分化,设诱导组和空白对照组,7d后通过微管相关蛋白2(MAP-2)免疫荧光及Hoechst标记观察海马神经干细胞向神经元的分化情况。结果正常组和对照组条件培养基中CLU浓度较低,约为(0.1037±0.0119)ng/L和(0.1170±0.0078)ng/L;切割组12h、24h、48h条件培养基中其浓度明显增高,分别为(0.1940±0.0364)ng/L、(0.4250±0.0724)ng/L和(0.2903±0.0472)ng/L,以24h组最高,约为正常组和对照组的4倍。CLU诱导组中MAP-2阳性细胞数明显高于空白对照组(P0.05),其占总细胞数的百分比分别为(11.67±2.57)%和(4.52±1.54)%,且诱导组中MAP-2阳性细胞突起更长、更丰富。结论切割穹隆海马伞侧海马提取液"激活"的星形胶质细胞可分泌更多的CLU,体外细胞培养中CLU可促进海马神经干细胞向成熟的神经元分化。
[Abstract]:Objective to investigate the "activation" of astrocytes secreting aggregin protein (CLU) from the lateral hippocampal extract of the fimbria fornix and the differentiation of neural stem cells into neurons induced by CLU in vitro. Methods astrocytes were cultured in DMEM / F12 medium with the hippocampal extract cut from the fimbria fornix, and cultured in the normal hippocampal extract DMEM / F12 medium. The concentration of CLU protein in different conditioned media was detected by Elisa in normal group and control group, and the differentiation of neural stem cells in hippocampus was induced by CLU in vitro. The differentiation of hippocampal neural stem cells into neurons was observed by microtubule-associated protein 2 (MAP-2) immunofluorescence and Hoechst labeling after 7 days in induction group and blank control group. Results the concentrations of CLU in conditioned medium of normal group and control group were (0.1037 卤0.0119) ng / L and (0.1170 卤0.0078) ng / L, respectively, and the concentration of CLU in conditioned medium of 12h and 24h / 48h was (0.1940 卤0.0364) ng / L, (0.4250 卤0.0724) ngP / L and (0.2903 卤0.0472) ng / L, respectively, the highest was in 24 h group, and the highest in 24 h group, (0.4250 卤0.0724) ng / L and (0.2903 卤0.0472) ng / L, respectively. The number of MAP-2 positive cells in CLU induced group was significantly higher than that in blank control group (P0.05), and the percentage of MAP-2 positive cells in the total cells was (11.67 卤2.57)% and (4.52 卤1.54), respectively. Conclusion the astrocytes "activated" in the hippocampal extract of fimbria incised fornix can secrete more CLU, and CLU can promote the differentiation of hippocampal neural stem cells into mature neurons in vitro.
【作者单位】：南通大学医学院人体解剖学系江苏省神经再生重点实验室;
【基金】：江苏省高校自然科学研究项目(10KJD180002) 江苏省高校优势学科建设工程资助项目(PAPD)
【分类号】：R329

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