莱菔硫烷抑制LPS诱导的小鼠星形胶质细胞增殖  

Sulforaphane inhibits astrocyte proliferation by induced LPS in mice

作　　者：陈亚星[1] 袁继超[1] 刘伟[1] 朱海涛[1] 廖小俊[1] 文泽贤 李兰[1] 林江凯[1] 

机构地区：[1]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,重庆400038

出　　处：《第三军医大学学报》2017年第5期407-412,共6页Journal of Third Military Medical University

基　　金：国家自然科学基金青年科学基金(81401012);国家自然科学基金面上项目(81571214)~~

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0.1μg/m L 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、20μmol/L)处理星形胶质细胞24 h后,CCK8法检测细胞增殖情况;选取20μmol/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、Cyclin D1、PCNA的表达情况。结果 CCK8法检测显示0.1μg/m L LPS能显著促进星形胶质细胞增殖(P<0.01),而加入20μmol/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20μmol/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20μmol/L莱菔硫烷能下调LPS刺激下PCNA、Cyclin D1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。astrocytes induced Objective To determine the effect by lipopolysaccharide （LPS）. Methods of sulforaphane （SFN） on the proliferation of 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, P27kipl, Cyclin D1 and proliferating cell nuclear antigen （PCNA） in each group of cells were determined by Western blotting. Results CCK8 assay indicated that O. 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 〈 O. 05 ）. Cell cycle analysis showed that 20 vmol/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 P27kipl （ P 〈 0.05 ）. Oonclusion SFN inhibits the proliferation of astrocytes induced by LPS, which may be through regulation of cell cycle.

关 键 词：莱菔硫烷 星形胶质细胞 LPS 细胞周期 细胞增殖 

分 类 号：R322.8[医药卫生—人体解剖和组织胚胎学] R363[医药卫生—基础医学]