本文關(guān)鍵詞： 睪丸免疫豁免 自身免疫性睪丸炎 Toll樣受體 TAM受體酪氨酸激酶　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

【摘要】：背景與目的：哺乳動(dòng)物睪丸是一個(gè)典型的免疫豁免組織,系統(tǒng)性的免疫耐受和睪丸局部免疫抑制環(huán)境是維持免疫豁免的重要機(jī)制。生理狀態(tài)下,生精細(xì)胞抗原在睪丸中不誘導(dǎo)免疫反應(yīng),但病理?xiàng)l件下,這種免疫平衡會(huì)被破壞,引起自身免疫性睪丸炎。睪丸局部免疫豁免及自身免疫性睪丸炎的調(diào)節(jié)機(jī)制有待深入研究。Toll樣受體(TLR)在啟動(dòng)免疫反應(yīng)中發(fā)揮重要作用,并可以介導(dǎo)自身免疫反應(yīng),而Tyro3, Axl與Mer (TAM)受體酪氨酸激酶能抑制TLR信號(hào)通路。本論文研究TLR與TAM對(duì)小鼠實(shí)驗(yàn)性自身免疫性睪丸炎(EAO)的調(diào)節(jié)作用,以揭示其在維持睪丸免疫豁免中的機(jī)制。材料方法：用完全佐劑乳化的睪丸勻漿免疫小鼠,誘導(dǎo)EAO模型。HE染色分析睪丸組織結(jié)構(gòu)和損傷情況：免疫組化和流式細(xì)胞儀分析睪丸間質(zhì)中巨噬細(xì)胞及淋巴細(xì)胞浸潤(rùn)情況；qRT-PCR和ELISA分析睪丸中促炎癥因子表達(dá)；Western blot分析血清中自身抗體；生物素示蹤的方法分析睪丸BTB結(jié)構(gòu)完整性。結(jié)果：經(jīng)過一次免疫同源睪丸組織勻漿,Axl和Mer雙基因敲除(Ax1-/-Mer-/-)小鼠能誘導(dǎo)嚴(yán)重EAO。免疫50 d后,產(chǎn)生明顯的EAO特征。淋巴結(jié)腫大,產(chǎn)生明顯的系統(tǒng)性炎癥反應(yīng)；睪丸萎縮,重量減輕,生精上皮出現(xiàn)嚴(yán)重?fù)p傷：睪丸間質(zhì)中出現(xiàn)明顯的巨噬細(xì)胞和淋巴細(xì)胞浸潤(rùn)；睪丸內(nèi)促炎癥因子表達(dá)上調(diào)；血睪屏障被破壞；血清中產(chǎn)生抗生精細(xì)胞的自身抗體。經(jīng)過一次誘導(dǎo),Axl-/-和Mer-/-小鼠能產(chǎn)生相對(duì)弱的EAO,而在WT小鼠中不產(chǎn)生EAO。經(jīng)過三次免疫,WT小鼠可產(chǎn)生嚴(yán)重EAO,出現(xiàn)典型的EAO表型。TLR2和TLR4單基因敲除(TLR2-/-和TLR4-/-)小鼠能誘導(dǎo)相對(duì)弱的EAO,而TLR2和TLR4雙基因敲除(TLR2-/-TLR4-/-)小鼠不產(chǎn)生EAO。而且發(fā)現(xiàn)TLR2-/-和TLR2-/-TLR4-/-小鼠經(jīng)過誘導(dǎo)后血清中沒有產(chǎn)生自身抗體,表明TLR2在介導(dǎo)生殖細(xì)胞自身抗體產(chǎn)生中起關(guān)鍵作用。結(jié)論：TLR2和TLR4共同作用,介導(dǎo)EAO產(chǎn)生。而Axl和Mer協(xié)同作用,抑制EAO產(chǎn)生,這種抑制作用可能是通過抑制TLR2和TLR4信號(hào)通路實(shí)現(xiàn)的。研究結(jié)果進(jìn)一步揭示了系統(tǒng)性免疫反應(yīng)調(diào)節(jié)睪丸免疫豁免的機(jī)制。
[Abstract]:Background & objective: the mammalian testis are a typical immune immunity free tissue. Systemic immune tolerance and local immunosuppressive environment of testis are important mechanisms to maintain immunity immunity. Spermatogenic cell antigens do not induce immune responses in the testis, but this immune balance can be disrupted under pathological conditions. The local immunity immunity of testis and the regulatory mechanism of autoimmune orchitis need to be further studied. Toll-like receptor TLRR plays an important role in initiating the immune response. And can mediate autoimmune response while Tyro3. Axl and Mer tam receptor tyrosine kinase can inhibit TLR signaling pathway. In this study, we studied the regulatory effects of TLR and TAM on experimental autoimmune orchitis in mice. In order to reveal its mechanism in maintaining testicular immunity immunity. Material method: mice were immunized with complete adjuvant emulsified testis homogenate. EAO model. HE staining was used to analyze the structure and injury of testis: immunohistochemistry and flow cytometry were used to analyze the infiltration of macrophages and lymphocytes in testis stroma. QRT-PCR and ELISA were used to analyze the expression of proinflammatory factors in testis. Serum autoantibodies were analyzed by Western blot. The structural integrity of testicular BTB was analyzed by biotin tracer. Results: homologous homologous testis homogenate was immunized. Axl and Mer double gene knockout ax1-r-Mer-r-mice could induce severe EAO. After 50 days of immunization, they produced obvious EAO characteristics and enlarged lymph nodes. To produce obvious systemic inflammatory reaction; Testicular atrophy, weight reduction, spermatogenic epithelium appeared serious damage: clear interstitial macrophages and lymphocyte infiltration; The expression of proinflammatory factor was up-regulated in testis. The blood-testis barrier was destroyed; Autoantibodies against spermatogenic cells are produced in the serum. After one induction, axl-r-r-and Mer-r-mice produce relatively weak EAOs, whereas in WT mice, EAOs are not produced. WT mice can produce severe EAO, typical EAO phenotype. TLR2 and TLR4 single gene knockout TLR2-r- and TLR4-r-) mice can induce relatively weak EAO. And TLR2 and TLR4 double gene knockout TLR2-r-TLR4-r-). Mice did not produce EAO. and no autoantibodies were found in the serum of TLR2-r-r- and TLR2-r-TLR4-r- induced mice. It is suggested that TLR2 plays a key role in mediating the production of germ cell autoantibodies. Conclusion: TLR2 and TLR4 act together to mediate the production of EAO, while Axl and Mer play a synergistic role. Inhibition of EAO production may be achieved by inhibiting TLR2 and TLR4 signaling pathways. The results further reveal the mechanism of systemic immune response in regulating testicular immune immunity.
【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)】：R392

【共引文獻(xiàn)】

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