脂多糖刺激小胶质细胞激活分化的机制研究
吴军;丁单华;李倩倩;王欣宇;孙玉莹;李兰珺
【期刊名称】《中华神经医学杂志》 【年(卷),期】2018(017)012
【摘要】目的 探讨脂多糖(LPS)对小胶质细胞激活分化的影响及其可能机制.方法 将体外常规培养的BV2小胶质细胞分为LPS组和对照组,LPS组细胞加入200 ng/mL LPS,对照组细胞加入等量培养基.作用6 h后应用酶联免疫吸附实验(ELISA)和实时荧光定量PCR(qRT-PCR)分别检测2组细胞培养液上清中肿瘤坏死因子 α(TNF-α)、白介素(IL)-1β 浓度和细胞TNF-α、IL-1βmRNA的表达;应用免疫荧光染色和qRT-PCR分别检测细胞形态、 一氧化氮合酶(iNOS)、Arg1、CD32、CD206蛋白和mRNA的表达;应用Western blotting和qRT-PCR分别检测2组细胞Notch1、Hes1和Hes5蛋白和mRNA的表达.结果 与对照组比较,LPS组细胞上清液中IL-1β、TNF-α的浓度和细胞IL-1β、TNF-αmRNA的表达均明显增高,差异有统计学意义(P<0.05);免疫荧光染色检测显示LPS组细胞被激活,形态呈现为类阿米巴样.与对照组比较,LPS组细胞iNOS、CD32蛋白和mRNA的表达增高,差异有统计学意义(P<0.05);2组细胞Arg1、CD206蛋白和mRNA的表达差异无统计学意义(P>0.05).与对照组比较,LPS组细胞Notch1和Hes1蛋白和mRNA的表达明显增加,差异有统计学意义(P<0.05);2组细胞Hes5蛋白和mRNA的表达差异无统计学意义(P>0.05).结论 LPS可能通过Notch信号通路调节小胶质细胞向M1型分化,促进炎症反应.Notch信号通路可能是调控小胶质细胞激活分化进而减轻炎症损伤的作用靶点.%Objective To observe the effect and molecular mechanism of
lipopolysaccharide (LPS) on activation and differentiation of microglia (MG) cells. Methods Routinely in vitro cultured BV2 microglia cells were divided into control group and LPS group: BV2 microglia cells in the LPS group were treated with 200 ng/mL LPS; cells in the control group were added the same amount of medium. Six h after treatment, real-time quantitative (qRT)-PCR and enzyme-linked immunosorbent assay (ELISA) were used to detect the inflammatory factors, interleukin (IL)-1β and tumor necrosis factor (TNF)-α mRNA and protein expressions in supernatant of cell culture medium. The iNOS, CD32, Arg1 and CD206 mRNA and protein expressions were detected by qRT-PCR and immunofluorescence, respectively. The mRNA and protein expressions of Notch1, Hes1 and Hes5 were detected by qRT-PCR and Western blotting. Results After LPS stimulation, BV2 microglia cells were activated and the morphological changes were observed. The IL-1β and TNF-α protein and mRNA expressions in the LPS group were significantly increased as compared with those in the control group (P<0.05). The iNOS and CD32 protein and mRNA expressions in the LPS group were significantly increased as compared with those in the control group (P<0.05). The Arg1 and CD206 mRNA and protein expressions showed no significant differences between the two groups (P>0.05). The Notch1 and Hes1 mRNA and protein expressions in the LPS group were significantly increased as compared with those in the
control group (P<0.05), while no significant differences on Hes5 mRNA and protein expressions were noted between the two groups (P>0.05). Conclusion LPS activates MG cells, which may regulate the differentiation of MG cells into M1 through Notch signaling pathway and promote inflammatory response; therefore, Notch signaling pathway may be a target for regulating MG cells differentiation and reducing inflammatory damage. 【总页数】8页(1195-1202)
【关键词】炎症反应;小胶质细胞;细胞分化;脂多糖;Notch信号通路 【作者】吴军;丁单华;李倩倩;王欣宇;孙玉莹;李兰珺
【作者单位】450052 郑州,郑州大学第一附属医院神经内科;450052 郑州,郑州大学第一附属医院神经内科;450052 郑州,郑州大学第一附属医院神经内科;450052 郑州,郑州大学第一附属医院神经内科;450052 郑州,郑州大学第一附属医院神经内科;450052 郑州,郑州大学第一附属医院神经内科 【正文语种】中文 【中图分类】R742 【文献来源】
https://www.zhangqiaokeyan.com/academic-journal-cn_chinese-journal-neuromedicine_thesis/0201270494438.html 【相关文献】
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脂多糖刺激小胶质细胞激活分化的机制研究
