本文選題：Orai1 + 變應性鼻炎��； 參考：《復旦大學》2010年博士論文

【摘要】： 目的：Orai1是組成鈣離子釋放激活的鈣離子通道(CRAC通道)至關重要的亞單位,對存儲調(diào)控性鈣離子內(nèi)流(SOCE)起著關鍵性作用。關于Orai1的研究目前已比較深入,但對于此通路在變應性鼻炎方面的作用目前尚未有研究涉及。本實驗意欲檢測Orai1在變應性鼻炎小鼠模型中的表達以及評估Orai1通路的阻滯對免疫球蛋白E (IgE)介導的過敏反應的影響。 材料和方法：采用BALB/c品系小鼠,通過重復腹腔內(nèi)注射卵清蛋白致敏、反復鼻腔內(nèi)滴入卵清蛋白激惹建立變應性鼻炎小鼠模型,然后用免疫組化染色法、蛋白印跡法(western blotting)和實時反轉(zhuǎn)錄聚合酶鏈反應法(real-time RT-PCR)檢測Orai1在對照組、實驗組和2-氨基乙酯二苯基硼酸(2-APB)干預組小鼠鼻粘膜和鼻相關淋巴組織(NALT)中的表達,另外用酶聯(lián)免疫吸附實驗(ELISA)法分析上述幾組小鼠鼻腔灌洗液中的白細胞三烯C4(LTC4)、嗜酸性粒細胞陽離子蛋白(ECP)、卵清蛋白特異性免疫球蛋白E (sIgE)和白細胞介素4(IL-4)的含量,并用real-time RT-PCR法檢測幾組小鼠鼻粘膜中的白細胞三烯C4合酶(LTC4S)信使核糖核酸(mRNA)、ECP mRNA的含量和小鼠NALT中的胚系Cε轉(zhuǎn)錄體、IL-4 mRNA的表達。 結果：研究發(fā)現(xiàn)Orai1在鼻粘膜上皮細胞、粘膜下腺上皮細胞和NALT的免疫系統(tǒng)細胞中均有表達,在實驗組表達尤為顯著；2-APB鼻內(nèi)滴入干預組小鼠相對于實驗組小鼠的噴嚏數(shù)、撓鼻次數(shù)以及鼻粘膜內(nèi)浸潤的嗜酸性粒細胞數(shù)都明顯減少,而且干預組Orai1在小鼠鼻粘膜和NALT中的表達也明顯少于實驗組,同時研究還發(fā)現(xiàn),干預組小鼠鼻腔灌洗液中的LTC4、ECP、slgE和IL-4以及這些物質(zhì)相應的mRNA的含量相對于實驗組亦有顯著抑制。 結論：本實驗證實Orai1在小鼠鼻粘膜和NALT中確有表達,2-APB小鼠鼻內(nèi)滴入的干預措施可以有效地緩解小鼠變應性鼻炎的發(fā)作。
[Abstract]:Objective: Orai1 is a critical subunit of the CRAC channel, which is a calcium channel activated by calcium release. It plays a key role in the storage regulation of calcium ion influx (SOCEE). The research on Orai1 has been deeply studied, but the role of this pathway in allergic rhinitis has not been studied. The purpose of this study was to detect the expression of Orai1 in allergic rhinitis mice and to evaluate the effect of Orai1 pathway block on immunoglobulin E (IgE) -mediated allergic reaction. Materials and methods: the allergic rhinitis mouse model was established by repeated intraperitoneal injection of ovalbumin sensitized by repeated intraperitoneal injection of ovalbumin and repeated instillation of ovalbumin to induce allergic rhinitis in BALB/c strain mice. Western blotting and real-time RT-PCR were used to detect the expression of Orai1 in the nasal mucosa and naso-associated lymphoid tissue of mice in control group, experimental group and 2-APB intervention group. In addition, the contents of leukocyte triene C4Tc 4, eosinophil cationic protein (ECPP), ovalbumin specific immunoglobulin E (IgE) and interleukin-4 (IL-4) in the nasal lavage fluid of the above mentioned groups were analyzed by Elisa. The real-time RT-PCR method was used to detect the content of mRNA of leukocyte triene C4 synthase (LTC4S) in mouse nasal mucosa and the expression of IL-4 mRNA in mouse NALT. Results: the expression of Orai1 was found in nasal epithelial cells, submucosal gland epithelial cells and immune system cells of NALT, especially in experimental group. The number of nasal scratches and the number of eosinophils infiltrated in the nasal mucosa were significantly decreased, and the expression of Orai1 in the nasal mucosa and NALT in the intervention group was significantly lower than that in the experimental group. The contents of LTC4 ECPG E and IL-4 and their corresponding mRNA in nasal lavage fluid in the intervention group were also significantly inhibited compared with the experimental group. Conclusion: this study confirmed that the intervention of Orai1 expression in nasal mucosa and NALT could effectively relieve allergic rhinitis in mice.
【學位授予單位】：復旦大學
【學位級別】：博士
【學位授予年份】：2010
【分類號】：R765.21

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本文編號：1880981