解剖学报

解剖学报 ›› 2018, Vol. 49 ›› Issue (2): 137-142.doi: 10.16098/j.issn.0529-1356.2018.02.001

• 神经生物学 •    下一篇

脑脂结合蛋白基因沉默抑制体外大鼠星形胶质细胞增殖

李浩明 杨清清 彭敏 秦建兵 韩笑 成翔 金国华*   

  1. 南通大学医学院人体解剖学系，神经生物学研究所，炎症微环境协同创新中心， 江苏 南通 226001
  • 收稿日期:2017-03-27 修回日期:2017-05-05 出版日期:2017-04-06 发布日期:2018-04-06
  • 通讯作者: 金国华 E-mail:jguohua@ntu.edu.cn
  • 基金资助:
    国家自然科学基金资助项目;江苏省自然科学基金资助项目;江苏省高等学校大学生创新创业训练计划项目

Silencing brainlipid binding protein gene inhibiting the proliferation of rat astrocytes in vitro

LI Hao-ming YANG Qing-qing PENG Min QIN Jiang-bing HAN Xiao CHENG Xiang JIN Guo-hua*   

  1. Department of Anatomy, Institute of Neurobiology, Collaborative Innovation Center of Inflammatory Microenviroment, Medical School, Nantong University， Jiangsu Nantong 226001, China
  • Received:2017-03-27 Revised:2017-05-05 Online:2017-04-06 Published:2018-04-06
  • Contact: JIN Guo-hua E-mail:jguohua@ntu.edu.cn

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摘要：

目的 探讨脑脂结合蛋白（BLBP）对体外培养的星形胶质细胞增殖能力的影响。 方法 在体外培养大鼠星形胶质细胞的基础上，应用小干扰RNA（siRNA）沉默细胞内源性BLBP的表达后，Real-time PCR和Western blotting分别检测BLBP及c-Myc基因和蛋白的表达变化；随后采用细胞计数试剂盒8（CCK-8）试剂检测细胞活力，Ki67免疫荧光标记增殖状态的细胞，并用流式细胞术检测细胞周期的变化。 结果 应用siRNA沉默星形胶质细胞内源性BLBP的表达后，BLBP基因和蛋白的表达量明显下降，原癌基因c-Myc的表达水平也显著下调；BLBP基因沉默能够抑制细胞的活力并减少Ki67阳性细胞的数量；流式细胞术检测后发现，BLBP表达不足能够减少S期细胞的比例。 结论 BLBP基因沉默能够抑制体外培养的星形胶质细胞的增殖能力，BLBP在星形胶质细胞的增殖过程中发挥调控作用。

关键词: 脑脂结合蛋白, 星形胶质细胞, 增殖, 小干扰RNA, 免疫印迹法, 大鼠

Abstract:

Objective To explore the function of brain-lipid binding protein (BLBP) in cultured rat astrocytes in vitro. Methods After culturing rat astrocytes, BLBP small interference RNA (siRNA) was used to silence the expression of endogenous BLBP. Real-time PCR and Western blotting were applied to detect the level of BLBP and c-Myc; the cells viability was detected by using CCK-8 regent, the proliferated cells were labeled by Ki67, and the cells cycle was investigated by flow cytometry. Results The expressions of BLBP and c-Myc were significantly decreased after treatment of BLBP siRNA. BLBP gene silencing suppressed the cells viability and decreased the proportion of Ki67-positive cells. The result of flow cytometry showed that the insufficient expression of BLBP reduced the cells number of S phase. Conclusion The gene silencing of BLBP inhibits the cells proliferation of astrocytes in vitro. BLBP may regulate the proliferation process of astrocytes.

Key words: Brain-lipid binding protein, Astrocyte, Proliferation, Small interference RNA, Western blotting, Rat