GM-CSF对肥大细胞蛋白酶激活受体表达和介质分泌的影响及其信号转导通路探讨

发布时间：2018-02-14 09:33

本文关键词： 肥大细胞蛋白酶激活受体(PARs) GM-CSF IL-4　出处：《汕头大学》2008年硕士论文　论文类型：学位论文

【摘要】： 背景与目的:蛋白酶激活受体(protease activated receptors,PARs),白细胞介素(interleukin,IL) IL-4,IL-6参与过敏反应的发生,粒-巨噬细胞集落刺激因子(granulocyte-macrophage colony-stimulating factor,GM-CSF)具有调节免疫应答的功能,然而GM-CSF对肥大细胞分泌细胞因子的影响还不甚清楚。因此本研究探讨GM-CSF对肥大细胞P815 PARs表达和IL-4,IL-6分泌的影响,并探讨可能的信号转导通路。方法:肥大细胞P815激发后,收集各时间点的细胞和上清。细胞处理后采用实时定量PCR和流式细胞术的方法在mRNA和蛋白水平检测PAR-1,PAR-2,PAR-3和PAR-4表达情况,用细胞激发信号ELISA(cellular activation of signaling ELISA,CASE)方法检测信号转导通路蛋白Akt,ERK,p38和STAT3磷酸化情况;上清用ELISA方法检测IL-4,IL-6水平。结果:GM-CSF上调PAR-2蛋白和mRNA在肥大细胞P815上的表达;GM-CSF上调PAR-1,PAR-3和PAR-4 mRNA在肥大细胞P815的表达;GM-CSF抗体的应用可以阻断GM-CSF对PARs蛋白表达的调节作用。GM-CSF以浓度依赖的方式促进肥大细胞P815分泌IL-4和IL-6;应用PD98059, U0126和LY294002可以阻断GM-CSF引起的肥大细胞IL-4和IL-6分泌并抑制其引起的ERK和Akt磷酸化。结论:GM-CSF能够调节肥大细胞PARs表达并刺激肥大细胞分泌IL-4和IL-6。GM-CSF引起肥大细胞P815分泌细胞因子很可能是通过激活Akt和ERK信号转导通路实现的,这些发现提示GM-CSF可以条件依赖性的调节Th1细胞因子和/或Th2细胞因子分泌,从而在过敏性炎症中发挥重要作用。
[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.
【学位授予单位】：汕头大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392

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