[1]陈亚星,袁继超,刘伟,等.莱菔硫烷抑制LPS诱导的小鼠星形胶质细胞增殖[J].第三军医大学学报,2017,39(05 ):407-412.
 Chen Yaxing,Yuan Jichao,Liu Wei,et al.Sulforaphane inhibits astrocyte proliferation by induced LPS in mice[J].J Third Mil Med Univ,2017,39(05 ):407-412.
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莱菔硫烷抑制LPS诱导的小鼠星形胶质细胞增殖(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
39卷
期数:
2017年第05期
页码:
407-412
栏目:
基础医学
出版日期:
2017-03-15

文章信息/Info

Title:
Sulforaphane inhibits astrocyte proliferation by induced LPS in mice
作者:
陈亚星袁继超刘伟朱海涛廖小俊文泽贤李兰林江凯
第三军医大学西南医院神经外科,全军神经外科研究所,全军神经创伤防治重点实验室
Author(s):
Chen Yaxing Yuan Jichao Liu Wei Zhu Haitao Liao Xiaojun Wen Zexian Li Lan Lin Jiangkai

Department of Neurosurgery, Institute of Neurosurgery, Key Laboratory of Neurotrauma Prevention and Treatment, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China

关键词:
莱菔硫烷星形胶质细胞LPS细胞周期细胞增殖
Keywords:
sulforaphane astrocyte LPS cell cycle cell proliferat
分类号:
R322.8; R363; R979.5
文献标志码:
A
摘要:

目的       探讨莱菔硫烷(sulforaphane,SFN)对脂多糖(LPS)诱导的星形胶质细胞(astrocyte,AST)增殖的影响及其机制。方法      体外培养小鼠原代星形胶质细胞,给予0.1 μg/mL LPS刺激星形胶质细胞活化增殖;同时分别给予不同浓度的莱菔硫烷(10、15、20 μmol/L)处理星形胶质细胞24 h后,CCK8法检测细胞增殖情况;选取20 μmol/L作为干预剂量行GFAP、Ki67双标免疫荧光染色及流式细胞仪细胞周期检测,进一步验证莱菔硫烷对LPS诱导星形胶质增殖的作用;Western blot检测细胞周期相关蛋白P27kip1、CyclinD1、PCNA的表达情况。结果      CCK8法检测显示0.1 μg/mL LPS能显著促进星形胶质细胞增殖(P<0.01),而加入20 μmol/L莱菔硫烷能显著抑制LPS诱导的星形胶质细胞增殖(P<0.01),并能减少LPS刺激下的Ki67阳性细胞(P<0.05);细胞周期分析提示20 μmol/L莱菔硫烷可减少LPS诱导下的星形胶质细胞S期比例,增加G1期细胞比例(P<0.05);Western blot提示20 μmol/L莱菔硫烷能下调LPS刺激下PCNA、CyclinD1的表达,上调P27kip1表达(P<0.05)。结论     莱菔硫烷可以通过调控星形胶质细胞细胞周期,抑制体外LPS诱导的星形胶质细胞增殖。

Abstract:

Objective     To determine the effect of sulforaphane (SFN) on the proliferation of astrocytes induced by lipopolysaccharide (LPS). Methods     Primary astrocytes were isolated from the mouse cortex and then cultured in vitro. After the cells were stimulated with 0.1 μg/mL LPS in the absence or presence of different concentrations of SFN (10, 15 and 20 μmol/L) for 24 h, the cell proliferation was tested with CCK8 assay. Immunofluorescent double staining and cell cycle analysis were performed respectively in the cells treated with 20 μmol/L SFN to further verify the effect of SFN on the proliferation of astrocytes after LPS stimulation. The expression levels of cell cycle related proteins, P27kip1, Cyclin D1 and proliferating cell nuclear antigen (PCNA) in each group of cells were determined by Western blotting. Results      CCK8 assay indicated that 0.1 μg/mL LPS promoted the proliferation of astrocytes (P<0.01). Treatment of 20 μmol/L SFN inhibited the proliferation significantly (P<0.01) and reduced the number of Ki67 positive cells under LPS stimulation (P<0.05). Cell cycle analysis showed that 20 μmol/L SFN treatment attenuated the increment of cells in S phase and the reduction of cells in G1 phase which were induced by LPS stimulation (P<0.05). Western blot assay represented that LPS enhanced the expression of PCNA and Cyclin D1, meanwhile, decreased the level of P27kip1(P<0.05). Conclusion      SFN inhibits the proliferation of astrocytes induced by LPS, which may be through regulation of cell cycle.

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更新日期/Last Update: 2017-03-09