中枢神经系统来源的HMGB1表达模式和释放特征
Expression Patterns and Release Characteristics of CNS-Derived HMGB1
投稿时间:2018-04-18  修订日期:2018-06-14
DOI:
中文关键词: 高迁移率族蛋白 1  星形胶质细胞  小胶质细胞  神经元
英文关键词: High mobility group box 1  astrocyte  microglial  neuron
基金项目:国家自然科学基金资助项目(81601046),中南民族大学大学生创新创业训练计划项目资助项目(GCX1724),中央高校基本科研业务费专项资金资助项目(CZQ16010,CZQ18010)
作者单位E-mail
孙燕 中南民族大学 生命科学学院武汉神经科学与神经工程研究所 sunyan198112@mail.scuec.edu.cn 
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中文摘要:
      目的:阐明生理病理状态下,中枢神经系统(CNS)来源的高迁移率族蛋白1(HMGB1)细胞定位、亚细胞定位和释放特征. 方法:使用Western Blot和细胞免疫荧光双标记法检测HMGB1在细胞株(U87, BV2和N2a)和原代星形胶质细胞、神经元的表达模式和释放特征. 用PTX和/或ATP处理培养细胞,Western Blot检测浓缩上清中HMGB1蛋白表达,台盼蓝拒染法检测细胞的生存率. 结果:HMGB1定位于U87, BV2, N2a,原代星形胶质细胞和神经元的细胞核,位于目的条带25 KDa位置. PTX和/或ATP能诱导U87和BV2主动分泌HMGB1,N2a细胞被动释放HMGB1. 结论:生理状态下,星形胶质细胞、小胶质细胞和神经元细胞核表达HMGB1,属于HMGB1来源,HMGB1具有CNS释放特征.
英文摘要:
      Objective :To investigate the cell location, subcellular location and release characteristics of CNS-derived HMGB1 under different pathophysiological status. Methods: Western Blot assay and double immunofluorescence staining were used to detect the expression patterns and release characteristics of HMGB1 from cell lines (U87, BV2 and N2a) and primary cultured astrocytes and neurons. Cells were cultured and stimulated with ATP alone or PTX followed by ATP . The expression of HMGB1 in the concentrated supernatant was measured by Western Blot and the cell survival rate was determined by trypan blue dye exclusion method. Results: U87, BV2, N2a, primary cultured astrocytes and neurons extract confirmed expression of a 25 kDa protein that was nuclear localization of HMGB1. With PTX and/or plus ATP stimulation, U87 and BV2 could actively release HMGB1, while N2a could passively release HMGB1. Conclusion: The nuclei of astrocytes, microglia and neurons can express HMGB1 under physiological state, suggesting that they are important resources of HMGB1 and HMGB1 has CNS release characteristics.
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