本文選題：半胱氨酰白三烯受體 + BV2小膠質(zhì)細(xì)胞��； 參考：《浙江大學(xué)》2014年碩士論文

【摘要】：目的：觀察半胱氨酰白三烯(CysLT)受體(CysLT1R和CysLT2R)對(duì)小鼠BV2小膠質(zhì)細(xì)胞吞噬功能的調(diào)節(jié)作用,并初步探討CysLT受體兩種亞型之間是否存在相互作用。 方法： 1.采用CysLT受體激動(dòng)劑(LTD4和NMLTC4)和小膠質(zhì)細(xì)胞激活劑(LPS和IFN-γ)作用一定時(shí)間,以免疫熒光法和流式細(xì)胞術(shù)檢測BV2細(xì)胞吞噬功能的變化。 2. CysLT1R選擇性拮抗劑montelukast和CysLT2R選擇性拮抗劑HAMI3379預(yù)處理30min后,檢測對(duì)激動(dòng)劑或激活劑引起的吞噬功能變化的作用。 3.以Western blotting法檢測LPS和IFN-y引起的CysLT1R及CysLT2R表達(dá)的變化,以及montelukast和HAMI3379對(duì)LPS引起的表達(dá)變化的影響。 4.以免疫熒光共染法檢測(?)CysLT受體激動(dòng)劑LTD4和NMLTC4以及LPS是否引起CysLT1R和CysLT2R分布的變化；在確定的作用時(shí)間點(diǎn),用激光共聚焦顯微鏡進(jìn)行初步驗(yàn)證。 結(jié)果： 1.免疫熒光法和流式細(xì)胞術(shù)都表明,CysLTs受體激動(dòng)劑LTD4和NMLTC4以及小膠質(zhì)細(xì)胞激活劑LPS和IFN-γ,均增強(qiáng)BV2細(xì)胞吞噬功能。 2. Montelukast和HAMI3379能對(duì)抗BV2細(xì)胞吞噬功能的增強(qiáng)。Montelukast(0.1μmol/L)和HAMI3379(1μmol/L)均顯著抑制LTD4誘導(dǎo)的吞噬增強(qiáng)；montelukast (0.1-1μmol/L)和HAMI3379(1μmol/L)均明顯抑制NMLTC4誘導(dǎo)的吞噬增強(qiáng),且montelukast作用強(qiáng)于HAMI3379. Montelukast和HAMI3379(1μmol/L)均顯著抑制小膠質(zhì)細(xì)胞激活劑LPS誘導(dǎo)的吞噬增強(qiáng)。 3.LPS和IFN-γ均上調(diào)BV2細(xì)胞的CysLT1R和CysLT2I表達(dá),其中,IFN-y使CysLT1I表達(dá)僅有增加趨勢,而CysLT2I表達(dá)明顯增加；LPS顯著增加CysLT1R和CysLT2R的表達(dá),且該作用能被montelukast和HAMI3379所拮抗。 4.BV2細(xì)胞激活導(dǎo)致CysLT1R和CysLT2R分布的變化,CysLT1R與CysLT2I存在共定位,激活后發(fā)生內(nèi)移的特點(diǎn)與時(shí)間基本一致。 結(jié)論： 1.CysLT1R和CysLT2R共同參與BV2細(xì)胞激活的調(diào)節(jié),表現(xiàn)為細(xì)胞吞噬功能增強(qiáng)以及CysLT1R和CysLT2I滾達(dá)增加,特異性拮抗劑使用后上述現(xiàn)象被逆轉(zhuǎn)。 2. CysLT1R和CysLT2R之間可能存在的相互作用,但需進(jìn)一步研究確定。
[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- 緯) 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- 緯 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- 緯 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.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R964

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相關(guān)期刊論文 前3條

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本文編號(hào)：1856441