| 摘要: |
| 目的：了解高表达糖原合成酶激酶3β（GSK3β）对巨噬细胞功能的影响.方法：构建pcDNA3.1-GSK3β真核表达载体,脂质体转染小鼠巨噬细胞RAW264.7,G418筛选获得GSK3β高表达细胞系;采用中性红染色法、Griess法和结晶紫染色法分别测定巨噬细胞吞噬功能、细胞上清中一氧化氮（NO）含量和肿瘤坏死因子（TNF）活性.结果：小鼠巨噬细胞系RAW264.7高表达GSK3β,经酶切鉴定和测序证明一致,能抑制巨噬细胞吞噬功能（抑制率20.4%,P＜0.05）和脂多糖（LPS, 1 μg/ml）诱导的巨噬细胞NO的产生（抑制率25.6%,P＜0.01）,促进TNF的产生（增加率28.8%,P＜0.01）.结论：GSK3β具有调节巨噬细胞参与炎症免疫反应的功能. |
| 关键词: 糖原合成酶激酶3β 巨噬细胞 吞噬作用 一氧化氮 肿瘤坏死因子 |
| DOI：10.3724/SP.J.1008.2007.00697 |
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| 基金项目:国家自然科学基金（30472032）. |
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| Influence of glycogen synthase kinase 3β overexpression on function of murine macrophage RAW264.7 |
| LI Yuan-yuan , WANG Shuo-feng , ZHANG Yue-fan , XIAO Zhen-yu , ZHANG Jun-ping , WAN Li-li , GUO Cheng. |
| (1. Department of Biochemical Pharmacy, School of Pharmacy, Second Military Medical University, Shanghai 200433 ,China ; 2. Department of Pharmacy, Sixth People; s Hospital of Shanghai, Shanghai Jiaotong University, Shanghai 200233) |
| Abstract: |
| Objective： To study the influence of glycogen synthase kinase 3β（GSK3β） overexpression on the function of murine macrophage RAW264.7. Methods： Eucaryotic vector pcDNA3. 1-GSK3β was constructed and transfected into RAW264.7 cells via lipofectamine 2000. RAW264.7 cells stably expressing GSK3β were screened by G418 selection. Phagocytosis ability of RAW264.7 cells was assessed by neutral red analysis; the content of nitric oxide （NO） and the activity of tumor necrosis factor （TNF） in the supernatant of RAW264. 7 cells stably expressing GSK3β were determined by Griess method and crystal violet staining assay, respectively. Results： RAW264.7 cells stably expressing GSK3β showed an inhibited phagocytosis ability,with the inhibitory rate being 20.4% （P〈0. 05）, and an inhibited production of NO by RAW264.7 cells stably expressing GSK3β in presence of 1 μg/ml LPS, with the inhibitory rate being 25. 6%（P〈0. 01）. The transfected RAW264.7 cells had an increased production of TNF by 28.8% （P〈0. 01）. Conclusion： GSK3β can regulate macrophages in inflammatory and immune responses. |
| Key words: glycogen synthase kinase 3β macrophages phagocytosis nitric oxide tumor necrosis factor |
