白三烯D_4体外诱导小胶质细胞激活的作用  

Leukotriene D_4 activates BV2 microglia in vitro

作　　者：张壮[1] 骆江云[1] 黄静[1] 刘志娴[1] 方三华[1] 张纬萍[1] 魏尔清[1] 卢韵碧[1] 

机构地区：[1]浙江大学医学院药理学系卫生部医学神经生物学重点实验室,浙江杭州310058

出　　处：《浙江大学学报（医学版）》2013年第3期253-260,共8页Journal of Zhejiang University(Medical Sciences)

基　　金：国家自然科学基金(81173041;81072619;81072618);浙江省科技厅钱江人才项目(2010R10055);中央高校基本科研业务费专项资金(2011QNA7015)

摘　　要：目的:观察白三烯D4(LTD4)激活小胶质细胞株BV2细胞的作用,探讨介导BV2细胞激活的半胱氨酰白三烯受体(CysLT受体)亚型。方法:以Western blotting和免疫组化鉴定BV2细胞上CysLT受体的表达;以LTD4及CysLT受体拮抗剂处理BV2细胞后,采用MTT还原法检测细胞活性,微珠示踪法(以流式细胞仪检测)观察细胞吞噬的变化,RT-PCR检测炎症因子IL-6 mRNA的表达变化。结果:LTD4不影响BV2细胞增殖,但随着LTD4浓度增高可增强细胞吞噬、增加IL-6mRNA的表达,100 nmol/L LTD4使细胞吞噬增高为对照的1.4倍(P<0.001),IL-6 mRNA的表达增高为对照的2倍(P<0.01);LTD4诱导BV2细胞上调表达IL-6 mRNA可被CysLT1受体选择性拮抗剂孟鲁司特和CysLT1/CysLT2非选择性拮抗剂BAY u9773抑制,而CysLT2受体选择性拮抗剂HAMI 3379对此无明显影响;HAMI 3379和BAY u9773(100 nmol/L)能进一步增强LTD4诱导的BV2细胞吞噬。结论:体外LTD4可激活BV2细胞,LTD4诱导细胞IL-6 mRNA上调表达由CysLT1受体介导,LTD4诱导的细胞吞噬增强受CysLT2受体负性调节。Objective： To investigate the effects of CysLT receptor agonist leukotriene D4（LTD4） and antagonists on activation of microglia BV2 cells.Methods： The expression of CysLT1 and CysLT2 protein was determined by Western blotting and immunostaining in microglia BV2 cells.BV2 cells were pretreated with or without CysLT1 receptor selective antagonist montelukast,CysLT2 receptor selective antagonist HAMI 3379,or CysLT1/CysLT2 receptor dual antagonist BAY u9773 for 30 min,then the cells were treated with LTD4 for 24 h.Cell viability was detected by MTT reduction assay.Phagocytosis and mRNA expression of IL-6 were determined by fluorescent bead tracking and RT-PCR,respectively.Results： In BV2 cells,LTD4 did not affect proliferation but significantly enhanced phagocytosis and increased IL-6 mRNA expression in a concentration-dependent manner.LTD4 at 100 nmol/L induced a 1.4-fold increase of phagocytic index and a 2-fold up-regulation of IL-6 mRNA expression（P0.01）.HAMI 3379 and BAY u9773（100 nmol/L） further increased LTD4-induced phagocytosis;BAY u9773 and montelukast decreased LTD4-induced IL-6 mRNA expression,while HAMI 3379 had no effect on that.Conclusion： LTD4 activates BV2 cells in vitro and enhances IL-6 mRNA expression mediated by CysLT1 receptor,LTD4 induces phagocytosis which might be negatively regulated by CysLT2 receptor in BV2 cells.

关 键 词：白三烯D4 小胶质细胞 半胱氨酸 受体 白三烯 吞噬作用 细胞因子 

分 类 号：R363[医药卫生—病理学]