半胱氨酰白三烯受体对BV2细胞吞噬功能的调节作用

发布时间：2018-05-07 09:48

  本文选题：半胱氨酰白三烯受体 + BV2小胶质细胞　； 参考：《浙江大学》2014年硕士论文

【摘要】：目的：观察半胱氨酰白三烯(CysLT)受体(CysLT1R和CysLT2R)对小鼠BV2小胶质细胞吞噬功能的调节作用,并初步探讨CysLT受体两种亚型之间是否存在相互作用。 方法： 1.采用CysLT受体激动剂(LTD4和NMLTC4)和小胶质细胞激活剂(LPS和IFN-γ)作用一定时间,以免疫荧光法和流式细胞术检测BV2细胞吞噬功能的变化。 2. CysLT1R选择性拮抗剂montelukast和CysLT2R选择性拮抗剂HAMI3379预处理30min后,检测对激动剂或激活剂引起的吞噬功能变化的作用。 3.以Western blotting法检测LPS和IFN-y引起的CysLT1R及CysLT2R表达的变化,以及montelukast和HAMI3379对LPS引起的表达变化的影响。 4.以免疫荧光共染法检测(?)CysLT受体激动剂LTD4和NMLTC4以及LPS是否引起CysLT1R和CysLT2R分布的变化；在确定的作用时间点,用激光共聚焦显微镜进行初步验证。 结果： 1.免疫荧光法和流式细胞术都表明,CysLTs受体激动剂LTD4和NMLTC4以及小胶质细胞激活剂LPS和IFN-γ,均增强BV2细胞吞噬功能。 2. Montelukast和HAMI3379能对抗BV2细胞吞噬功能的增强。Montelukast(0.1μmol/L)和HAMI3379(1μmol/L)均显著抑制LTD4诱导的吞噬增强；montelukast (0.1-1μmol/L)和HAMI3379(1μmol/L)均明显抑制NMLTC4诱导的吞噬增强,且montelukast作用强于HAMI3379. Montelukast和HAMI3379(1μmol/L)均显著抑制小胶质细胞激活剂LPS诱导的吞噬增强。 3.LPS和IFN-γ均上调BV2细胞的CysLT1R和CysLT2I表达,其中,IFN-y使CysLT1I表达仅有增加趋势,而CysLT2I表达明显增加；LPS显著增加CysLT1R和CysLT2R的表达,且该作用能被montelukast和HAMI3379所拮抗。 4.BV2细胞激活导致CysLT1R和CysLT2R分布的变化,CysLT1R与CysLT2I存在共定位,激活后发生内移的特点与时间基本一致。 结论： 1.CysLT1R和CysLT2R共同参与BV2细胞激活的调节,表现为细胞吞噬功能增强以及CysLT1R和CysLT2I滚达增加,特异性拮抗剂使用后上述现象被逆转。 2. CysLT1R和CysLT2R之间可能存在的相互作用,但需进一步研究确定。
[Abstract]:Aim: to investigate the effects of cysteinyl leukotriene receptor (CysLT1R) and CysLT2R (CysLT2R) on phagocytosis of BV2 microglia in mice, and to explore whether there is interaction between the two subtypes of CysLT1R and CysLT2R. Methods: 1. The changes of phagocytosis of BV2 cells were detected by immunofluorescence and flow cytometry with CysLT receptor agonists (LTD4 and NMLTC4) and microglial activators (LPs and IFN- 纬) for a certain time. 2. The effects of CysLT1R selective antagonist montelukast and CysLT2R selective antagonist HAMI3379 on phagocytosis induced by agonist or activator were detected after 30min was pretreated. 3. The changes of CysLT1R and CysLT2R expression induced by LPS and IFN-y and the effect of montelukast and HAMI3379 on LPS expression were detected by Western blotting assay. 4. The distribution of CysLT receptor agonists LTD4, NMLTC4 and LPS were detected by immunofluorescence co-staining, and the distribution of CysLT1R and CysLT2R was preliminarily verified by laser confocal microscopy at a certain time point. Results: 1. Immunofluorescence assay and flow cytometry showed that CysLTs receptor agonists LTD4 and NMLTC4 and microglial activators LPS and IFN- 纬 enhanced the phagocytosis of BV2 cells. 2. Montelukast and HAMI3379 could inhibit the enhancement of phagocytosis induced by LTD4 (0.1 渭 mol / L) and HAMI3379(1 (0.1 渭 mol / L) and HAMI3379(1 渭 mol / L (0.1-1 渭 mol / L), and montelukast was stronger than HAMI3379. Both Montelukast and HAMI3379(1 渭 mol / L significantly inhibited the enhancement of phagocytosis induced by microglial activator LPS. Both 3.LPS and IFN- 纬 upregulated the expression of CysLT1R and CysLT2I in BV2 cells. The expression of CysLT1I increased only by IFN-y, while the expression of CysLT2I significantly increased the expression of CysLT1R and CysLT2R, which was antagonized by montelukast and HAMI3379. Activation of 4.BV2 cells resulted in changes in the distribution of CysLT1R and CysLT2R. CysLT1R was colocated with CysLT2I, and the characteristics of inward migration after activation were basically consistent with the time. Conclusion: 1.CysLT1R and CysLT2R were involved in the regulation of the activation of BV2 cells, including the enhancement of phagocytosis and the increase of CysLT1R and CysLT2I. The above phenomenon was reversed after the use of specific antagonists. 2. The possible interaction between CysLT1R and CysLT2R needs further study and determination.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R964

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