癌变·畸变·突变 ›› 2015, Vol. 27 ›› Issue (5): 375-377,382.doi: 10.3969/j.issn.1004-616x.2015.05.010

• 论著 • 上一篇    下一篇

微囊藻毒素-LR所致神经小胶质细胞BV-2的炎性反应

张敏辉, 周珏, 刘冉, 李晓波, 梁戈玉, 李云晖   

  1. 东南大学公共卫生学院环境医学工程教育部重点实验室, 江苏 南京 210009
  • 收稿日期:2015-03-13 修回日期:2015-05-05 出版日期:2015-09-30 发布日期:2015-09-30
  • 通讯作者: 李云晖,E-mail:lyh1216@126.com E-mail:lyh1216@126.com
  • 作者简介:张敏辉,Tel:15805157216;E-mail:zhangminhui1990@163.com。
  • 基金资助:

    国家自然科学基金(81072258);江苏省自然科学基金(BK2009281)

Effect of mycrocystin-LR on inflammatory response in BV-2 cells

ZHANG Minhui, ZHOU Jue, LIU Ran, LI Xiaobo, LIANG Geyu, LI Yunhui   

  1. Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Public Health, Southeast University, Nanjing 210009, Jiangsu, China
  • Received:2015-03-13 Revised:2015-05-05 Online:2015-09-30 Published:2015-09-30

摘要:

目的:通过检测微囊藻毒素-LR(MC-LR)对BV-2细胞内一氧化氮(NO)和一氧化氮合酶(NOS),以及培养上清液中的肿瘤坏死因子-α(TNF-α)和白介素-6(IL-6)表达的影响,为进一步探讨MC-LR引起中枢神经系统免疫反应奠定基础。方法:用MC-LR刺激BV-2细胞建立炎症模型,以四甲基噻唑蓝(MTT)法测定MC-LR对细胞的毒性作用;采用硝酸还原酶法检测BV-2细胞中NO的表达量;NOS分型法检测NOS的表达水平;酶联免疫吸附实验(ELISA)检测TNF-α和IL-6炎症因子的产生情况。结果:MC-LR染毒24 h后,染毒浓度在0~128 μg/L范围内,随着剂量的增加,细胞活性呈下降趋势(r=-0.609,P<0.05),其半数有效浓度(EC50)为22.49 μg/L。当染毒浓度≥5.0 μg/L时,细胞分泌的IL-6和NOS的含量明显增加,与对照组比较差异具有统计学意义(P<0.05);10.0 μg/L染毒后TNF-α和NO含量明显增加,与对照组比较差异具有统计学意义(P<0.05)。结论:MC-LR诱导BV-2细胞产生大量的炎性细胞因子NO、NOS、IL-6及TNF-α,引起神经系统的炎症反应。

关键词: 微囊藻毒素, BV-2细胞, 神经炎症, 一氧化氮, 一氧化氮合酶

Abstract:

OBJECTIVE: To investigate the effects of mycrocystin-LR(MC-LR) on the neurotoxic mechanisms of the central nervous system, the contents of nitric oxide (NO), nitric oxide synthase (NOS), tumor necrosis factor-α (TNF-α), and interleukin- 6 (IL-6) in BV-2 microglial cells were detected in our study. METHODS:With the methyl thiazolyl tetrazolium (MTT) assay, the cytotoxicity of the inflammatory model in BV-2 cells was performed after exposure to MC-LR. The levels of NO and NOS were measured by the nitric acid reductase assay and nitric oxide synthase assay, respectively. The contents of TNF-α and IL-6 were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS:After exposure to MC-LR for 24 h, the viability in BV-2 cells was decreased with the increase of MC-LR concentration within the range of 0-28 μg/L (r=-0.609, P<0.05), and the median effective concentration (EC50) was 22.49 μg/L. Besides, the levels of IL-6 and NOS in BV-2 cells were increased at the concentration of 5-10 μg/L MC-LR, and the levels of TNF-α and NO were increased at the concentration of 10 μg/L MC-LR. Compared with control group, those differences above mentioned were statistically significant (P<0.05). CONCLUSION:MC-LR induced inflammatory response in BV-2 microglial cells by increases in production of NO, NOS, IL-6 and TNF-α.

Key words: microcystin-LR, BV-2 cells, neuroinflammation, nitric oxide, nitric oxide synthase

中图分类号: