本文關(guān)鍵詞： 肥大細(xì)胞 蛋白酶激活受體(PARs) GM-CSF IL-4　出處：《汕頭大學(xué)》2008年碩士論文　論文類型：學(xué)位論文

【摘要】： 背景與目的:蛋白酶激活受體(protease activated receptors,PARs),白細(xì)胞介素(interleukin,IL) IL-4,IL-6參與過敏反應(yīng)的發(fā)生,粒-巨噬細(xì)胞集落刺激因子(granulocyte-macrophage colony-stimulating factor,GM-CSF)具有調(diào)節(jié)免疫應(yīng)答的功能,然而GM-CSF對肥大細(xì)胞分泌細(xì)胞因子的影響還不甚清楚。因此本研究探討GM-CSF對肥大細(xì)胞P815 PARs表達(dá)和IL-4,IL-6分泌的影響,并探討可能的信號轉(zhuǎn)導(dǎo)通路。 方法:肥大細(xì)胞P815激發(fā)后,收集各時間點(diǎn)的細(xì)胞和上清。細(xì)胞處理后采用實(shí)時定量PCR和流式細(xì)胞術(shù)的方法在mRNA和蛋白水平檢測PAR-1,PAR-2,PAR-3和PAR-4表達(dá)情況,用細(xì)胞激發(fā)信號ELISA(cellular activation of signaling ELISA,CASE)方法檢測信號轉(zhuǎn)導(dǎo)通路蛋白Akt,ERK,p38和STAT3磷酸化情況;上清用ELISA方法檢測IL-4,IL-6水平。 結(jié)果:GM-CSF上調(diào)PAR-2蛋白和mRNA在肥大細(xì)胞P815上的表達(dá);GM-CSF上調(diào)PAR-1,PAR-3和PAR-4 mRNA在肥大細(xì)胞P815的表達(dá);GM-CSF抗體的應(yīng)用可以阻斷GM-CSF對PARs蛋白表達(dá)的調(diào)節(jié)作用。GM-CSF以濃度依賴的方式促進(jìn)肥大細(xì)胞P815分泌IL-4和IL-6;應(yīng)用PD98059, U0126和LY294002可以阻斷GM-CSF引起的肥大細(xì)胞IL-4和IL-6分泌并抑制其引起的ERK和Akt磷酸化。 結(jié)論:GM-CSF能夠調(diào)節(jié)肥大細(xì)胞PARs表達(dá)并刺激肥大細(xì)胞分泌IL-4和IL-6。GM-CSF引起肥大細(xì)胞P815分泌細(xì)胞因子很可能是通過激活A(yù)kt和ERK信號轉(zhuǎn)導(dǎo)通路實(shí)現(xiàn)的,這些發(fā)現(xiàn)提示GM-CSF可以條件依賴性的調(diào)節(jié)Th1細(xì)胞因子和/或Th2細(xì)胞因子分泌,從而在過敏性炎癥中發(fā)揮重要作用。
[Abstract]:Background & AIM: protease activated receptor (activated receptor) IL-4IL-6 is involved in the development of allergic reaction. Granulocyte-macrophage colony-stimulating factor GM-CSF has the function of regulating immune response. However, the effect of GM-CSF on the secretion of cytokines by mast cells is not clear. Therefore, the effects of GM-CSF on the expression of P815 PARs and the secretion of IL-4- 6 in mast cells were investigated, and the possible signal transduction pathways were explored. Methods: after P815 of mast cells were stimulated, cells and supernatants were collected at different time points. The expression of PAR-1 and PAR-2PAR-3 and PAR-4 were detected by real-time quantitative PCR and flow cytometry at the level of mRNA and protein. The phosphorylation of the signal transduction pathway proteins Aktttr ERKN p38 and STAT3 was detected by ELISA(cellular activation of signaling CASEs, and the IL-4 IL-6 level was detected by ELISA method in the supernatant. Results GM-CSF upregulated the expression of PAR-2 protein and mRNA on mast cell P815. GM-CSF upregulated the expression of PAR-1, PAR-3 and PAR-4 mRNA in mast cell P815. The application of GM-CSF antibody could block the effect of GM-CSF on PARs protein expression. GM-CSF promoted PARs protein expression in a concentration-dependent manner. P815 secreted IL-4 and IL-6, and PD98059, U0126 and LY294002 blocked IL-4 and IL-6 secretion induced by GM-CSF and inhibited ERK and Akt phosphorylation induced by PD98059, U0126 and LY294002. ConclusionThe PARs expression of mast cells and the secretion of IL-4 and IL-6.GM-CSF by mast cells can be regulated by 1: GM-CSF and P815 cytokines secreted by mast cells may be mediated by activating the signal transduction pathways of Akt and ERK. These findings suggest that GM-CSF can conditionally regulate the secretion of Th1 cytokines and / or Th2 cytokines and thus play an important role in allergic inflammation.
【學(xué)位授予單位】：汕頭大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 耿志華;肺泡蛋白沉著癥發(fā)病機(jī)制及其治療的研究進(jìn)展[J];國外醫(yī)學(xué)(內(nèi)科學(xué)分冊);2002年09期

2 田從哲;杜錦朵;肖二彬;高永平;;53例鼻息肉及鼻息肉病患者組織GM-CSF、IL-5檢測[J];解放軍預(yù)防醫(yī)學(xué)雜志;2006年04期

3 邵春忠,李瑛,史玉民,肖小平,劉文彥;粒細(xì)胞-巨噬細(xì)胞集落刺激因子與α-干擾素聯(lián)合治療慢性乙型肝炎患者對細(xì)胞免疫的影響[J];臨床肝膽病雜志;2003年01期

4 朱雅群;張力元;田野;;重組人粒-巨噬細(xì)胞集落刺激因子治療放射性食管炎的療效[J];江蘇醫(yī)藥;2007年03期

5 房彤;高宏;陳力;孫保錦;;重組人粒-巨噬細(xì)胞集落刺激因子治療放射性口腔黏膜炎療效分析[J];中國腫瘤臨床與康復(fù);2006年06期

6 王選錠,羅發(fā)滿,劉富光,Burkhard Bewig;粒-巨噬細(xì)胞集落刺激因子在肺泡蛋白沉積癥患者中的表達(dá)[J];中華結(jié)核和呼吸雜志;2001年07期

7 周賢梅;白久武;蔡后榮;吳鴻雁;陳駿;;粒-巨噬細(xì)胞集落刺激因子及肺表面活性蛋白在肺泡蛋白沉積癥患者肺內(nèi)的表達(dá)[J];中華內(nèi)科雜志;2005年11期

8 陳興平,張勇,陳映玲;重組人粒細(xì)胞巨噬細(xì)胞集落刺激因子與伊曲康唑聯(lián)合對白念珠菌抗菌作用的實(shí)驗(yàn)研究[J];中華皮膚科雜志;2004年08期

9 石勝軍,肖能坎;蛋白酶激活受體家族與創(chuàng)面愈合[J];中華燒傷雜志;2003年04期

10 陳焰,焦志勇,田德英;乙型肝炎患者血清粒細(xì)胞-巨噬細(xì)胞集落刺激因子檢測的臨床意義[J];中西醫(yī)結(jié)合肝病雜志;2003年02期

，

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