本文選題：蛋白酶激活受體　切入點(diǎn)：肥大細(xì)胞　出處：《南方醫(yī)科大學(xué)》2010年碩士論文

【摘要】： 蛋白酶激活受體(protease activated receptors,PARs)是絲氨酸蛋白酶受體,目前在人和小鼠體內(nèi)共發(fā)現(xiàn)了四種PARs,分別為PAR-1、PAR-2、PAR-3和PAR-4。它們的基因序列于1991、1994、1997和1998年被先后克隆出來(lái)。其中PAR-1是凝血酶和胰蛋白酶的受體,PAR-2是胰蛋白酶、類(lèi)胰蛋白酶和彈性蛋白酶的受體,PAR-3是凝血酶受體,PAR-4是凝血酶和胰蛋白酶受體。 很久以來(lái)就認(rèn)識(shí)到肥大細(xì)胞分泌介質(zhì)參與過(guò)敏反應(yīng)。作為初級(jí)效應(yīng)細(xì)胞,肥大細(xì)胞既參與急性過(guò)敏反應(yīng)又參與慢性過(guò)敏反應(yīng)。肥大細(xì)胞一旦被激活,即釋放其已預(yù)先合成的和新產(chǎn)生的介質(zhì)執(zhí)行其生物學(xué)功能。這些介質(zhì)包括組胺、肝素、蛋白酶以及一系列的細(xì)胞因子如IL-4、IL-5和IL-6等,這些細(xì)胞因子可以在生理和病理情況下參與調(diào)節(jié)細(xì)胞的生長(zhǎng)、分泌和遷移等。 有關(guān)PARs與過(guò)敏反應(yīng)性疾病的研究發(fā)現(xiàn),PARs尤其是PAR-2與過(guò)敏反應(yīng)密切相關(guān)。一些過(guò)敏原如塵螨、蟑螂過(guò)敏原可以激活PAR-2。但通過(guò)疾病動(dòng)物模型得到的實(shí)驗(yàn)結(jié)果卻不盡相同：有些結(jié)果顯示PAR-2激活引起呼吸道的過(guò)敏性炎癥反應(yīng),有些則發(fā)現(xiàn)PAR-2的激活在過(guò)敏性疾病中起到保護(hù)作用。所以有待于進(jìn)一步深入探討PARs在過(guò)敏反應(yīng)性疾病中的作用。雖然對(duì)于參與過(guò)敏反應(yīng)的一些細(xì)胞如內(nèi)皮細(xì)胞、上皮細(xì)胞、中性粒細(xì)胞、單核細(xì)胞、淋巴細(xì)胞等的PARs功能已經(jīng)有所了解,然而對(duì)過(guò)敏性疾病的核心效應(yīng)細(xì)胞—肥大細(xì)胞的PARs功能還知之甚少。因此本研究將進(jìn)一步的探討肥大細(xì)胞PARs的功能,為過(guò)敏性疾病發(fā)病機(jī)制的研究提供實(shí)驗(yàn)數(shù)據(jù)。 一、蛋白酶激活受體在肥大細(xì)胞的表達(dá) 用逆轉(zhuǎn)錄聚合酶鏈?zhǔn)椒磻?yīng)、流式細(xì)胞術(shù)及免疫熒光細(xì)胞染色分析方法,在mRNA水平和蛋白水平檢測(cè)PAR-1、PAR-2、PAR-3和PAR-4在小鼠肥大細(xì)胞MC/9和P815的表達(dá)。結(jié)果小鼠肥大細(xì)胞MC/9和P815在mRNA水平和蛋白水平均表達(dá)PAR-1、PAR-2、PAR-3和PAR-4。結(jié)論肥大細(xì)胞MC/9和P815表達(dá)PAR-1、PAR-2、PAR-3和PAR-4。 二、絲氨酸蛋白酶對(duì)肥大細(xì)胞IL-4分泌的影響 肥大細(xì)胞P815培養(yǎng)后用胰蛋白酶、類(lèi)胰蛋白酶、凝血酶單獨(dú)或與相應(yīng)抑制劑凝血酶抑制劑、大豆胰蛋白酶抑制劑、α抗胰蛋白酶抑制劑、亮肽素結(jié)合處理肥大細(xì)胞,收集上清并用ELISA方法檢測(cè)IL-4的水平。結(jié)果：胰蛋白酶和類(lèi)胰蛋白酶能促進(jìn)肥大細(xì)胞P815分泌IL-4明顯增多(P0.05)；大豆胰蛋白酶抑制劑和α抗胰蛋白酶能阻斷胰蛋白酶引起的肥大細(xì)胞P815 IL-4分泌(P0.05)；亮肽素能阻斷類(lèi)胰蛋白酶引起的肥大細(xì)胞P815 IL-4分泌(P0.05),而凝血酶對(duì)肥大細(xì)胞P815分泌IL-4無(wú)明顯影響(P0.05)。結(jié)論：胰蛋白酶和類(lèi)胰蛋白酶刺激肥大細(xì)胞IL-4分泌,而凝血酶對(duì)肥大細(xì)胞IL-4的分泌功能無(wú)影響。
[Abstract]:Protease activated receptor (PARs) is a serine protease receptor. At present, four species of pars have been found in human and mouse. They are PAR-1, PAR-2, PAR-3 and PAR-4. their gene sequences were cloned in 199119941997 and 1998.The receptor of PAR-1 is thrombin and trypsin, PAR-2 is trypsin. Trypsin and elastase receptors PAR-3 are thrombin receptors PAR-4 are thrombin and trypsin receptors. It has long been recognized that mast cell secretory mediators are involved in allergic reactions. As primary effector cells, mast cells are involved in both acute and chronic allergic reactions. That is, release its presynthesized and newly produced media for its biological functions. These mediums include histamine, heparin, protease, and a series of cytokines such as IL-4, IL-5 and IL-6. These cytokines are involved in the regulation of cell growth, secretion and migration, both physiologically and pathologically. Studies on PARs and allergic reactivity diseases have found that PARs, especially PAR-2, are closely related to allergic reactions. Some allergens such as dust mites, Cockroach allergens activate PAR-2. But the results from animal models of disease vary: some of the results show that PAR-2 activates allergic inflammation in the respiratory tract. Some have found that the activation of PAR-2 plays a protective role in allergic diseases. Therefore, it is necessary to further explore the role of PARs in allergic diseases, although for some cells involved in allergic reactions, such as endothelial cells, epithelial cells, The PARs function of neutrophils, monocytes, lymphocytes, etc. However, little is known about the PARs function of mast cells, which is the core effector of allergic diseases. Therefore, this study will further explore the function of mast cell PARs in order to provide experimental data for the study of pathogenesis of allergic diseases. The first is the expression of protease activated receptor in mast cells. Reverse transcriptase polymerase chain reaction, flow cytometry and immunofluorescence staining were used. The expressions of PAR-1PAR-2PAR-3 and PAR-4 in mouse mast cell MC/9 and P815 were detected at the level of mRNA and protein. Results the expression of PAR-1PAR-2PAR-3 and PAR-4in mouse mast cell MC/9 and P815 at the mRNA level and protein level was detected. Conclusion the expression of PAR-1PAR-2PAR-2PAR-3 and PAR-4Mast cell in mast cell MC/9 and P815 were detected. Effect of serine protease on IL-4 secretion of mast cells. Mast cells P815 were treated with trypsin, trypsin, thrombin alone or with the corresponding inhibitor thrombin inhibitor, soybean trypsin inhibitor, 偽 -antitrypsin inhibitor, and leucopeptide binding. The levels of IL-4 were detected by ELISA. Results: trypsin and trypsin could increase the secretion of IL-4 in mast cells, and soybean trypsin inhibitor and 偽 -antitrypsin could block the secretion of IL-4 by trypsin. Leucopeptide could block the P815 IL-4 secretion of mast cells induced by trypsin, while thrombin had no significant effect on the IL-4 secretion of mast cells. Conclusion: trypsin and trypsin stimulate the secretion of IL-4 in mast cells. Thrombin had no effect on the secretory function of mast cell IL-4.
【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類(lèi)號(hào)】：R392

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 劉素菊;人蛋白酶活化受體-2在子癇前期胎盤(pán)組織中的表達(dá)及其臨床意義[D];吉林大學(xué);2011年

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本文編號(hào)：1670432