【摘要】：背景與目的：多種病毒可以感染卵巢,影響卵巢的功能。一些病毒可通過感染卵細(xì)胞傳播給子代。然而針對病毒感染的卵巢天然免疫反應(yīng)尚未見報道。本論文旨在研究模式識別受體介導(dǎo)的卵巢天然抗病毒反應(yīng)及其對卵巢功能的影響。材料與方法：利用TLR3-/-和TNF-a-/-小鼠模型、結(jié)合RNA干擾技術(shù)研究相關(guān)基因的功能。使用免疫組化和Western blot方法分析蛋白的定位與定量表達。Real-time RT-PCR定量分析基因mRNA的水平。ELISA用來測定細(xì)胞因子及雌二醇的濃度。利用poly(I:C)處理細(xì)胞與小鼠模擬病毒感染,分析卵巢的天然抗病毒反應(yīng)。結(jié)果：病毒RNA模式識別受體TLR3、MDA5和RIG-I在卵巢顆粒細(xì)胞和基質(zhì)細(xì)胞中表達。其共同配體poly(I:C)可以誘導(dǎo)卵巢細(xì)胞的天然抗病毒反應(yīng),包括誘導(dǎo)免疫調(diào)節(jié)因子TNF-α, IL-6、IFN-α和IFN-β以及抗病毒蛋白OAS1、ISG15和MX1的表達。Poly(I:C)誘導(dǎo)的卵巢天然抗病毒反應(yīng)需要TLR3、MDA5、RIG-I受體介導(dǎo)的核轉(zhuǎn)錄因子κB(NF-κB)與干擾素調(diào)節(jié)因子3(IRF3)的活化。Poly(I:C)顯著抑制雌激素的合成,并誘導(dǎo)有腔卵泡的顆粒細(xì)胞凋亡,從而干擾卵泡成熟。然而TLR3-/-與TNF-α-/-小鼠的卵巢功能不受poly(I:C)影響,說明poly(I:C)主要通過TLR3介導(dǎo)產(chǎn)生的TNF-α損傷卵巢的功能。結(jié)論：卵巢顆粒細(xì)胞和基質(zhì)細(xì)胞表達模式識別受體TLR3、MDA5和RIG-I,并可介導(dǎo)天然抗病毒反應(yīng)。卵巢的天然抗病毒反應(yīng)抑制雌激素合成、誘導(dǎo)顆粒細(xì)胞凋亡、干擾卵泡成熟。TLR3介導(dǎo)產(chǎn)生的TNF-a是影響卵巢功能的關(guān)鍵因子。揭示了小鼠卵巢細(xì)胞天然抗病毒反應(yīng)及其干擾卵巢功能的機理。
[Abstract]:Background & objective: multiple viruses can infect ovary and affect ovarian function. Some viruses can be transmitted to offspring by infecting eggs. However, ovarian innate immune response to viral infection has not been reported. The aim of this study was to study the natural antiviral response of ovary mediated by pattern recognition receptor and its effect on ovarian function. Materials and methods: TLR3-/- and TNF-a-/- mouse models were used to study the function of related genes with RNA interference technique. Immunohistochemical and Western blot methods were used to analyze the localization and quantitative expression of protein. Real-time RT-PCR was used to quantitatively analyze the level of gene mRNA. ELISA was used to determine the concentration of cytokines and estradiol. Poly (I: C) was used to treat cells and mice to simulate virus infection and to analyze the natural antiviral response of ovary. Results: TLR3,MDA5 and RIG-I were expressed in granulosa cells and stromal cells of ovary. Its co-ligand poly (I: C) can induce natural antiviral responses in ovarian cells, including the induction of immunomodulatory factors TNF- 偽, IL-6,IFN- 偽 and IFN- 尾 and the antiviral protein OAS1,. Expression of ISG15 and MX1. Poly (I: C) induced natural ovarian antiviral response requires TLR3,MDA5, RIG-I receptor mediated nuclear transcription factor 魏 B (NF- 魏 B) and activation of interferon regulatory factor 3 (IRF3). Poly (I: C significantly inhibited estrogen synthesis and induced apoptosis of granulosa cells with antral follicles, which interfered with follicular maturation. However, the ovarian function of TLR3-/- and TNF- 偽-/-mice is not affected by poly (I: C), which indicates that poly (I: C) mainly damages ovarian function by TNF- 偽 mediated by TLR3. Conclusion: the expression pattern recognition receptors TLR3,MDA5 and RIG-I, of granulosa cells and stromal cells of ovary can mediate natural antiviral response. The natural antiviral response of ovary inhibits estrogen synthesis, induces granulosa cell apoptosis and interferes follicular maturation. TNF-a mediated by TLR3 is the key factor affecting ovarian function. The natural antiviral response of mouse ovarian cells and its mechanism of interfering with ovarian function were revealed.
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：R392

【參考文獻】

相關(guān)期刊論文 前1條

1 ;An update on primary ovarian insufficiency[J];Science China(Life Sciences);2012年08期

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本文編號：2409902