本文關(guān)鍵詞： 慢性鼻-鼻竇炎 鼻息肉 DNA依賴的干擾素調(diào)節(jié)因子激活物 雙鏈DNA BEAS-2B細(xì)胞系 體外培養(yǎng) I型干擾素 促炎因子 抗炎因子 負(fù)反饋調(diào)節(jié)　出處：《華中科技大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

【摘要】：第一部分 胞內(nèi)DNA感受器DAI在慢性鼻-鼻竇炎和鼻息肉組織中的表達(dá) 目的 研究DNA依賴的干擾素調(diào)節(jié)因子激活物(DNA-dependent activator of IFN-regulatory factors,DAI) mRNA以及蛋白在慢性鼻-鼻竇炎、鼻息肉和正常下鼻甲鼻黏膜組織的表達(dá)及表達(dá)程度的差異,探討其在慢性鼻-鼻竇炎、和鼻息肉發(fā)病機(jī)制中的作用。 方法 RT-PCR檢測DAI在人類支氣管上皮細(xì)胞系BEAS-2B細(xì)胞及慢性鼻-鼻竇炎、鼻息肉和正常下鼻甲鼻黏膜組織的表達(dá)。real time RT-PCR技術(shù)和免疫組織化學(xué)法(IHC)分別檢測慢性鼻-鼻竇炎、鼻息肉和下鼻甲黏膜組織中DAI的表達(dá)水平。 結(jié)果 DAI mRNA及蛋白在BEAS-2B細(xì)胞、慢性鼻-鼻竇炎、慢性鼻-鼻竇炎合并鼻息肉和下鼻甲組織中均有表達(dá),real-time RT-PCR及免疫組化顯示慢性鼻-鼻竇炎和鼻息肉組織DAI的表達(dá)水平明顯低于下鼻甲組織,而以慢性鼻-鼻竇炎組織中下降更為明顯。 結(jié)論 DAI mRNA及蛋白在慢性鼻-鼻竇炎、鼻息肉和正常下鼻甲黏膜上皮細(xì)胞內(nèi)均有表達(dá),且表達(dá)量不同,差異有統(tǒng)計學(xué)意義,提示DAI可能在慢性鼻-鼻竇炎的發(fā)病機(jī)制中起重要作用。 第二部分 雙鏈DNA對BEAS-2B細(xì)胞及體外培養(yǎng)的鼻粘膜組織塊的影響 目的 1觀察雙鏈DNA分子poly(dA-dT).poly(dT-dA)是否能夠誘導(dǎo)BEAS-2B細(xì)胞產(chǎn)生I型干擾素及DAI。 2了解dsDNA刺激BEAS-2B細(xì)胞后所產(chǎn)生各細(xì)胞因子的影響,探討其相互作用。 3使用鼻粘膜組織塊體外培養(yǎng)研究dsDNA對鼻粘膜組織中相關(guān)細(xì)胞因子的影響,探討DNA受體DAI分子在慢性鼻-鼻竇炎、鼻息肉急性復(fù)發(fā)中的作用。 方法 1 poly(dA-dT).poly(dT-dA)轉(zhuǎn)染BEAS-2B細(xì)胞,real time RT-PCR檢測相關(guān)細(xì)胞因子的表達(dá)水平,western blot檢測DAI的表達(dá)。 2 dsDNA轉(zhuǎn)染體外培養(yǎng)的鼻粘膜組織塊,real time RT-PCR檢測相關(guān)細(xì)胞因子的表達(dá)水平。 結(jié)果 1 poly(dA-dT).poly(dT-dA)能夠誘導(dǎo)BEAS-2B細(xì)胞產(chǎn)生I型干擾素IFN-β,且具有時間與劑量依賴性。dsDNA刺激BEAS-2B細(xì)胞后產(chǎn)生的應(yīng)答中,早期主要表現(xiàn)為促炎因子的升高,后期則以抗炎因子的高表達(dá)為主,且dsDNA對抗炎因子的誘導(dǎo)更為明顯。 2 dsDNA刺激體外培養(yǎng)的鼻粘膜組織塊后,表現(xiàn)為IL-6、IL-8、IP-10、IL-1β及MCP-3的高表達(dá)主要出現(xiàn)在NP中;TNF-α、IL-17、IL-22及IFN-λ2的高表達(dá)主要出現(xiàn)CRS中;而正常下鼻甲粘膜卻對dsDNA的反應(yīng)性較低。 結(jié)論 用dsDNA刺激BEAS-2B細(xì)胞后,機(jī)體通過自身負(fù)反饋調(diào)節(jié),最終表現(xiàn)以抗炎作用為主,對機(jī)體起保護(hù)性作用。在病原微生物刺激后,慢性鼻-鼻竇炎、鼻息肉比正常下鼻甲粘膜組織可能更容易受到影響,導(dǎo)致慢性鼻-鼻竇炎及鼻息肉的急性發(fā)作。而上述這些作用,可能正是通過dsDNA的受體DAI發(fā)揮作用的。
[Abstract]:Part one Expression of intracellular DNA receptor DAI in chronic rhinosinusitis and nasal polyps Purpose To study the DNA dependent activator of IFN-regulatory factors. To investigate the expression of mRNA and protein in chronic rhinosinusitis, nasal polyp and normal inferior turbinate nasal mucosa. And the role of nasal polyps in pathogenesis. Method DAI was detected by RT-PCR in human bronchial epithelial cell line BEAS-2B and chronic rhinosinusitis. Real time RT-PCR technique and immunohistochemical method were used to detect chronic rhinosinusitis. Expression of DAI in nasal polyps and inferior turbinate mucosa. Results DAI mRNA and protein were expressed in BEAS-2B cells, chronic rhinosinusitis, chronic rhinosinusitis with nasal polyps and inferior turbinate. Real-time RT-PCR and immunohistochemistry showed that the expression of DAI in chronic rhinosinusitis and nasal polyps was significantly lower than that in inferior turbinate. The decrease in chronic rhinosinusitis was more obvious. Conclusion DAI mRNA and protein were expressed in the epithelial cells of chronic rhinosinusitis, nasal polyps and normal inferior turbinate mucosa. The results suggest that DAI may play an important role in the pathogenesis of chronic rhinosinusitis. Part two Effects of double-stranded DNA on BEAS-2B cells and nasal mucosal tissue mass cultured in vitro Purpose 1. To observe whether the double-stranded DNA molecule polydA-dTU. PolydT-dA) can induce the production of type I interferon and dai in BEAS-2B cells. 2 to investigate the effects of dsDNA on the cytokines produced by BEAS-2B cells. 3The effect of dsDNA on cytokines in nasal mucosa was studied in vitro by using nasal mucosal tissue mass, and the expression of DNA receptor DAI molecule in chronic rhinosinusitis was studied. The role of nasal polyps in acute recurrence. Method 1 the expression of cytokines in BEAS-2B cells was detected by real time RT-PCR. The expression of DAI was detected by western blot. 2 the expression of cytokines was detected by real time RT-PCR in nasal mucosa transfected with dsDNA in vitro. Results (1) BEAS-2B cells were induced to produce IFN- 尾 type I interferon. In the response of BEAS-2B cells stimulated by dsDNA in a time-and dose-dependent manner, the high expression of anti-inflammatory factors was predominant in the early stage and in the later stage. The induction of dsDNA anti-inflammatory factor was more obvious. (2) after stimulation of nasal mucosa by dsDNA in vitro, the high expression of IL-6, IL-8, IP-10, IL-1 尾 and MCP-3 were mainly found in NP; The high expression of IL-22 and IFN- 位 2 in TNF- 偽 and IL-17 mainly occurred in CRS. However, the normal mucosa of inferior turbinate was less reactive to dsDNA. Conclusion After dsDNA was used to stimulate BEAS-2B cells, the body through its own negative feedback regulation, the final manifestation of anti-inflammatory effect, play a protective role on the body, after the stimulation of pathogenic microorganisms. Chronic rhinosinusitis and nasal polyps may be more likely to be affected than normal inferior turbinate mucosa, leading to acute episodes of chronic rhinosinusitis and nasal polyps. It may be through the dsDNA receptor DAI.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R765

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