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机构地区:[1]浙江省绍兴市人民医院 [2]浙江大学绍兴医院输血科,绍兴312000 [3]南通大学基础医学院免疫与微生物实验室,南通226001
出 处:《中国免疫学杂志》2013年第10期1033-1036,1041,共5页Chinese Journal of Immunology
摘 要:目的:探讨内毒素脂多糖(Lipopolysaccharide,LPS)对大鼠星形胶质细胞热休克蛋白A12B(Heat shock proteins A 12B,HSPA12B)表达变化和细胞定位的影响。方法:用不同浓度(0.01、0.1、1、10μg/ml)和同一浓度(1μg/ml)不同时间(1、2、4、6、8、12、24小时)的LPS刺激星形胶质细胞,分别用RT-PCR和Western blot法检测星形胶质细胞HSPA12B的mRNA和蛋白质水平表达情况,间接免疫荧光双标法检测HSPA12B在星形胶质细胞中的细胞定位。结果:用LPS 1μg/ml刺激1小时后,HSPA12B mRNA的表达增加,4小时表达活性最高。HSPA12B蛋白水平表达变化与mRNA水平变化相一致。免疫细胞化学证明LPS诱导星形胶质细胞HSPA12B的表达定位在胞核。结论:LPS可诱导星形胶质细胞HSPA12B表达上调。HSPA12B可作为一种炎症反应蛋白参与炎症反应过程。Objective :To explore the effect of lipopolysaccharide (LPS) on the changes of the expression and intracellular lo- cation of Heat shock proteins A 12B (HSPAI2B) in rat astrocytes. Methods: Astrocytes were treated with LPS at different concentra- tion (0.01, 0.1, 1, 10 μg/ml) and at the same concentration (1μg/ml) for different time (1,2, 4, 6, 8, 12, 24 h). With RT- PCR and Western blot measurement of HSPA12B, the levels of HSPA12B gene and protein expression were observed. Intracellular lo- cation of HSPA12B was detected under fluorescence microscope. Results: HSPA12B mRNA increased after 1 h exposure to LPS ( 1 μg/ml), and peaked at 4 h. And the HSPA12B protein expression was paralleled with the levels of mRNA. Immunocytoehemieal stai- ning indicated the expression of HSPA12B was located in the nuclear following LPS challenge. Conclusion: Bacterial causative agent, LPS could induce the expression of HSPA12B mRNA in rat astrocytes at transcription and protein level. HSPA12B served as an inflam- matory response molecule participating in the inflammatory process.
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