本文關(guān)鍵詞：模式識(shí)別受體介導(dǎo)的小鼠睪丸和附睪的天然抗病毒反應(yīng)　出處：《北京協(xié)和醫(yī)學(xué)院》2015年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： Leydig細(xì)胞 精子 附睪上皮細(xì)胞 天然抗病毒反應(yīng) 模式識(shí)別受體

【摘要】：背景與目的：病毒感染引起的睪丸炎和附睪炎,是破壞男性生育能力的重要因素。目前這兩個(gè)生殖器官的天然抗病毒免疫機(jī)制尚不清楚,本論文主要研究模式識(shí)別受體介導(dǎo)的睪丸與附睪天然抗病毒機(jī)制,為預(yù)防和治療相關(guān)疾病提供新線索。材料與方法：以小鼠為模型,用poly(I:C)和HSV60模擬病毒感染,利用實(shí)時(shí)定量RT-PCR檢測(cè)基因的mRNA水平,Western blot測(cè)定基因的蛋白水平,免疫組化染色定位蛋白的分布,ELISA檢測(cè)細(xì)胞因子和睪酮的分泌。利用TLR3基因敲除(TLR3-/-)小鼠和RNA干擾技術(shù)確證基因的功能。結(jié)果：睪丸Leydig細(xì)胞組成性表達(dá)病毒核酸受體MDA5、RIG-Ip204,其中MDA5也在圓形和長(zhǎng)形精子中表達(dá)。MDA5和RIG-I可以被poly(I:C)激活,p204可以被HSV60激活,誘導(dǎo)睪丸抗病毒反應(yīng)。附睪上皮細(xì)胞組成性表達(dá)多個(gè)病毒核酸受體,包括TLR3、RIG-I和DAI。p204與cGAS可以被HSV60誘導(dǎo)表達(dá)。TLR3和RIG-I可以被poly(I:C)激活,而DAI、p204 與 cGA S可以被HSV60激活,啟動(dòng)附睪天然抗病毒反應(yīng)。在睪丸及附睪細(xì)胞中,poly(I:C)和 HSV60都能誘導(dǎo)Ⅰ型干擾素和抗病毒蛋白ISG15、OAS1和MX1的表達(dá)。poly(I:C)可以顯著上調(diào)炎癥因子TNF-α和MCP-1的表達(dá),但HSV60并不影響TNF-a和MCP-1的表達(dá)。在TLR3-/-細(xì)胞中或用siRNA敲低RIG-I、DAI、p204或cGAS時(shí),抗病毒反應(yīng)明顯減弱,說明這幾個(gè)受體協(xié)同介導(dǎo)睪丸及附睪的天然抗病毒反應(yīng)。結(jié)論：在睪丸中,MDA5/RIG-I啟動(dòng)抗RNA病毒的反應(yīng),而p204介導(dǎo)抗DNA病毒的天然免疫反應(yīng)；在附睪中,TLR3與RIG-I介導(dǎo)抗RNA病毒的反應(yīng),DAI、p204和cGAS參與抗DNA病毒的反應(yīng)。以上結(jié)果說明睪丸與附睪組織特異細(xì)胞具有完善的天然抗病毒能力。
[Abstract]:Background & objective: orchitis and epididymitis caused by virus infection are important factors to destroy male fertility. At present, the natural antiviral immune mechanism of these two reproductive organs is not clear. This paper mainly studies the natural antiviral mechanism of testis and epididymis mediated by pattern recognition receptor, which provides a new clue for the prevention and treatment of related diseases. Materials and methods: mouse model. The viral infection was simulated by polymorphic I: C) and HSV60. The mRNA level of the gene was detected by real-time quantitative RT-PCR and the protein level of the gene was determined by Western blot. Immunohistochemical staining was used to locate the distribution of proteins. ELISA was used to detect the secretion of cytokines and testosterone. TLR3 gene knockout was used to remove TLR3 / -). Mouse and RNA interference techniques confirmed the function of the gene. Results: testicular Leydig cells expressed viral nucleic acid receptor MDA5 constitutively. RIG-Ip204, in which MDA5 is also expressed in round and long spermatozoa. MDA5 and RIG-I can be activated by polypeptide I: C) and p204 can be activated by HSV60. Induces testicular antiviral response. Epididymal epithelial cells constitutively express multiple viral nucleic acid receptors, including TLR3. RIG-I and DAI.p204 and cGAS can be induced by HSV60. TLR3 and RIG-I can be activated by polycyclic C) and DAI. P204 and cGA S can be activated by HSV60, initiating the natural antiviral reaction of epididymis, in testicular and epididymal cells. Polymorphic I: C) and HSV60 can induce interferon type I and antiviral protein ISG15. The expression of OAS1 and MX1 could significantly up-regulate the expression of TNF- 偽 and MCP-1. However, HSV60 did not affect the expression of TNF-a and MCP-1. In TLR3-r-cells or when siRNA was used to knock down RIG-Idae P204 or cGAS. The antiviral response was significantly weakened, indicating that these receptors co-mediated the natural antiviral response of testis and epididymis. Conclusion: MDA5 / RIG-I initiates the anti-viral response in testis. P204 mediated innate immune response against DNA virus. In epididymis, TLR3 and RIG-I mediate the response to RNA virus. P204 and cGAS are involved in the response to DNA virus. These results suggest that testicular and epididymal tissue specific cells have perfect natural antiviral ability.
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類號(hào)】：R698.2

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相關(guān)期刊論文 前1條

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本文編號(hào)：1440017