胞内DNA感受器DAI在慢性鼻—鼻窦炎及鼻息肉发病机制中作用的研究

发布时间：2018-01-26 01:53

本文关键词： 慢性鼻-鼻窦炎鼻息肉 DNA依赖的干扰素调节因子激活物双链DNA BEAS-2B细胞系体外培养 I型干扰素促炎因子抗炎因子负反馈调节　出处：《华中科技大学》2010年硕士论文　论文类型：学位论文

【摘要】：第一部分胞内DNA感受器DAI在慢性鼻-鼻窦炎和鼻息肉组织中的表达目的研究DNA依赖的干扰素调节因子激活物(DNA-dependent activator of IFN-regulatory factors,DAI) mRNA以及蛋白在慢性鼻-鼻窦炎、鼻息肉和正常下鼻甲鼻黏膜组织的表达及表达程度的差异,探讨其在慢性鼻-鼻窦炎、和鼻息肉发病机制中的作用。方法 RT-PCR检测DAI在人类支气管上皮细胞系BEAS-2B细胞及慢性鼻-鼻窦炎、鼻息肉和正常下鼻甲鼻黏膜组织的表达。real time RT-PCR技术和免疫组织化学法(IHC)分别检测慢性鼻-鼻窦炎、鼻息肉和下鼻甲黏膜组织中DAI的表达水平。结果 DAI mRNA及蛋白在BEAS-2B细胞、慢性鼻-鼻窦炎、慢性鼻-鼻窦炎合并鼻息肉和下鼻甲组织中均有表达,real-time RT-PCR及免疫组化显示慢性鼻-鼻窦炎和鼻息肉组织DAI的表达水平明显低于下鼻甲组织,而以慢性鼻-鼻窦炎组织中下降更为明显。结论 DAI mRNA及蛋白在慢性鼻-鼻窦炎、鼻息肉和正常下鼻甲黏膜上皮细胞内均有表达,且表达量不同,差异有统计学意义,提示DAI可能在慢性鼻-鼻窦炎的发病机制中起重要作用。第二部分双链DNA对BEAS-2B细胞及体外培养的鼻粘膜组织块的影响目的 1观察双链DNA分子poly(dA-dT).poly(dT-dA)是否能够诱导BEAS-2B细胞产生I型干扰素及DAI。 2了解dsDNA刺激BEAS-2B细胞后所产生各细胞因子的影响,探讨其相互作用。 3使用鼻粘膜组织块体外培养研究dsDNA对鼻粘膜组织中相关细胞因子的影响,探讨DNA受体DAI分子在慢性鼻-鼻窦炎、鼻息肉急性复发中的作用。方法 1 poly(dA-dT).poly(dT-dA)转染BEAS-2B细胞,real time RT-PCR检测相关细胞因子的表达水平,western blot检测DAI的表达。 2 dsDNA转染体外培养的鼻粘膜组织块,real time RT-PCR检测相关细胞因子的表达水平。结果 1 poly(dA-dT).poly(dT-dA)能够诱导BEAS-2B细胞产生I型干扰素IFN-β,且具有时间与剂量依赖性。dsDNA刺激BEAS-2B细胞后产生的应答中,早期主要表现为促炎因子的升高,后期则以抗炎因子的高表达为主,且dsDNA对抗炎因子的诱导更为明显。 2 dsDNA刺激体外培养的鼻粘膜组织块后,表现为IL-6、IL-8、IP-10、IL-1β及MCP-3的高表达主要出现在NP中;TNF-α、IL-17、IL-22及IFN-λ2的高表达主要出现CRS中;而正常下鼻甲粘膜却对dsDNA的反应性较低。结论用dsDNA刺激BEAS-2B细胞后,机体通过自身负反馈调节,最终表现以抗炎作用为主,对机体起保护性作用。在病原微生物刺激后,慢性鼻-鼻窦炎、鼻息肉比正常下鼻甲粘膜组织可能更容易受到影响,导致慢性鼻-鼻窦炎及鼻息肉的急性发作。而上述这些作用,可能正是通过dsDNA的受体DAI发挥作用的。
[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- 伪 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.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R765

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