發(fā)布時(shí)間：2018-03-06 00:35

  本文選題：Leydig細(xì)胞　切入點(diǎn)：TLR受體　出處：《北京協(xié)和醫(yī)學(xué)院》2011年博士論文　論文類型：學(xué)位論文

【摘要】：背景與目的：睪丸是免疫豁免組織,然而睪丸仍能有效的清除感染,說明睪丸自身具有抵抗病原體的機(jī)制。本論文的主要目的是研究Leydig細(xì)胞在睪丸中的天然免疫功能。睪丸巨噬細(xì)胞表現(xiàn)出較低的免疫活性,其機(jī)理尚不清楚,我們研究了Leydig細(xì)胞對(duì)巨噬細(xì)胞免疫反應(yīng)的調(diào)節(jié)。 材料與方法：利用C57/BL6小鼠為模型分離純化小鼠Leydig細(xì)胞。利用定量實(shí)時(shí)RT-PCR (Q-PCR)分析基因mRNA的水平。使用Western blotting和免疫組化染色研究蛋白的表達(dá)定位。ELISA檢測(cè)細(xì)胞因子產(chǎn)生和睪酮合成。利用基因敲除小鼠模型研究TAM受體在TLR介導(dǎo)天然免疫反應(yīng)中的作用。 結(jié)果：小鼠Leydig細(xì)胞中表達(dá)較高水平的TLR3和TLR4,以及較低水平的TLR2、TLR7、TLR9和TLR12,但不表達(dá)其它TLR。Leydig細(xì)胞中的TLR3和TLR4可被其受體激活,會(huì)引起核因子κB (NF-kB)和干擾素調(diào)節(jié)因子3(IRF3)的活化,進(jìn)而誘導(dǎo)該細(xì)胞分泌炎癥因子(IL-1β,IL-6和TNF-α)口I型干擾素(IFN-α及IFN-β),對(duì)入侵病原體產(chǎn)生免疫防御作用。為防止睪丸組織持續(xù)產(chǎn)生慢性炎癥反應(yīng),Leydig細(xì)胞中TLR3和TLR4受體的激活必須受到嚴(yán)格的調(diào)控。研究發(fā)現(xiàn)Leydig細(xì)胞中同時(shí)表達(dá)Axl和Mer受體,它們以協(xié)同的方式抑制TLR3和TLR4信號(hào)傳導(dǎo),調(diào)節(jié)Leydig細(xì)胞的天然免疫反應(yīng)。敲除Axl及Mer雙基因敲除(Axl-/-Mer-/-)小鼠的Leydig細(xì)胞中TLR3和TLR4激活水平顯著升高。Gas6是TAM受體的特異性配體,可以顯著抑制Leydig細(xì)胞中TLR-介導(dǎo)的NF-κB和IRF-3的激活及炎癥因子的表達(dá),而這種抑制效應(yīng)在Axl-/-Mer-/- Leydig細(xì)胞中則不存在。這些結(jié)果說明Axl和Mer受體酪氨酸激酶在Leydig細(xì)胞中以協(xié)同的方式負(fù)調(diào)控TLR信號(hào)。另外我們發(fā)現(xiàn)TLR3與TLR4的活化可以抑制Leydig細(xì)胞合成睪酮的能力。實(shí)驗(yàn)表明Leydig細(xì)胞可以抑制巨噬細(xì)胞的免疫反應(yīng)能力,可能對(duì)維持睪丸免疫平衡和免疫豁免狀態(tài)具有重要作用。 結(jié)論：Leydig細(xì)胞可以通過TLR3和TLR4的激活啟動(dòng)睪丸天然免疫反應(yīng)；TAM受體可以負(fù)調(diào)控Leydig細(xì)胞中TLR信號(hào)傳導(dǎo)；Leydig細(xì)胞可以抑制巨噬細(xì)胞的免疫反應(yīng)。這些結(jié)果表明Leydig細(xì)胞對(duì)睪丸抵抗入侵病原體,維持睪丸免疫平衡及免疫豁免狀態(tài)可能發(fā)揮重要作用。
[Abstract]:Background & objective: the testis are immune free tissue, but the testis can still effectively clear the infection. The main purpose of this paper is to study the innate immune function of Leydig cells in testis. We studied the regulation of macrophage immune response by Leydig cells. Materials and methods: mouse Leydig cells were isolated and purified by C57 / BL6 mouse model. Quantitative real-time RT-PCR Q-PCR was used to analyze the level of gene mRNA. Western blotting and immunohistochemical staining were used to study the expression localization of protein. Elisa was used to detect cytokine production. The role of TAM receptor in the innate immune response mediated by TLR was studied by gene knockout mouse model. Results: higher levels of TLR3 and TLR4, and lower levels of TLR2TLR7, TLR9 and TLR12 were expressed in mouse Leydig cells, but no expression of TLR3 and TLR4 in other TLR.Leydig cells could be activated by their receptors, which resulted in activation of nuclear factor- 魏 B (NF-kB) and interferon regulatory factor 3IRF3 (IFR3). Furthermore, the cells were induced to secrete inflammatory cytokines such as IL-1 尾, IL-6 and TNF- 偽, and IFN- 偽 and IFN- 尾 were induced to secrete IFN- 偽 and IFN- 尾. In order to prevent the sustained chronic inflammatory response of testicular tissues and the activation of TLR3 and TLR4 receptors in Leydig cells, IFN- 偽 and IFN- 尾 were induced to induce the secretion of IFN- 偽 and IFN- 尾. It was found that both Axl and Mer receptors were expressed in Leydig cells. They synergistically inhibited TLR3 and TLR4 signal transduction and regulated the innate immune response of Leydig cells. TLR3 and TLR4 activation levels in Leydig cells of Axl and Mer knockout mice were significantly increased. Gas6 was a specific ligand of TAM receptor. TLR- mediated activation of NF- 魏 B and IRF-3 and the expression of inflammatory factors in Leydig cells were significantly inhibited. These results suggest that Axl and Mer receptor tyrosine kinase negatively regulate TLR signal in Leydig cells in a synergistic manner. In addition, we found that the activation of TLR3 and TLR4 can inhibit Leydig fine. The ability of cells to synthesize testosterone. Experiments have shown that Leydig cells inhibit the immune response of macrophages. It may play an important role in maintaining testicular immune balance and immune immunity. Conclusion TLR3 and TLR4 can activate testicular innate immunoreactive tam receptor, which can negatively regulate the TLR signal transduction in Leydig cells and inhibit the immune response of macrophages. These results suggest that Leydig cells can inhibit the immune response of macrophages. The testicles resist invading pathogens, Maintaining testicular immune balance and immune immunity may play an important role.
【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】：2011
【分類號(hào)】：R392.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 Mahmoud Huleihel,Eitan Lunenfeld;Regulation of spermatogenesis by paracrine/autocrine testicular factors[J];Asian Journal of Andrology;2004年03期

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本文編號(hào)：1572616