模式识别受体启动的卵巢天然抗病毒反应及其对卵巢功能的影响
发布时间：2019-01-17 09:31
【摘要】：背景与目的：多种病毒可以感染卵巢,影响卵巢的功能。一些病毒可通过感染卵细胞传播给子代。然而针对病毒感染的卵巢天然免疫反应尚未见报道。本论文旨在研究模式识别受体介导的卵巢天然抗病毒反应及其对卵巢功能的影响。材料与方法：利用TLR3-/-和TNF-a-/-小鼠模型、结合RNA干扰技术研究相关基因的功能。使用免疫组化和Western blot方法分析蛋白的定位与定量表达。Real-time RT-PCR定量分析基因mRNA的水平。ELISA用来测定细胞因子及雌二醇的浓度。利用poly(I:C)处理细胞与小鼠模拟病毒感染,分析卵巢的天然抗病毒反应。结果：病毒RNA模式识别受体TLR3、MDA5和RIG-I在卵巢颗粒细胞和基质细胞中表达。其共同配体poly(I:C)可以诱导卵巢细胞的天然抗病毒反应,包括诱导免疫调节因子TNF-α, IL-6、IFN-α和IFN-β以及抗病毒蛋白OAS1、ISG15和MX1的表达。Poly(I:C)诱导的卵巢天然抗病毒反应需要TLR3、MDA5、RIG-I受体介导的核转录因子κB(NF-κB)与干扰素调节因子3(IRF3)的活化。Poly(I:C)显著抑制雌激素的合成,并诱导有腔卵泡的颗粒细胞凋亡,从而干扰卵泡成熟。然而TLR3-/-与TNF-α-/-小鼠的卵巢功能不受poly(I:C)影响,说明poly(I:C)主要通过TLR3介导产生的TNF-α损伤卵巢的功能。结论：卵巢颗粒细胞和基质细胞表达模式识别受体TLR3、MDA5和RIG-I,并可介导天然抗病毒反应。卵巢的天然抗病毒反应抑制雌激素合成、诱导颗粒细胞凋亡、干扰卵泡成熟。TLR3介导产生的TNF-a是影响卵巢功能的关键因子。揭示了小鼠卵巢细胞天然抗病毒反应及其干扰卵巢功能的机理。
[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- 伪, IL-6,IFN- 伪 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- 伪-/-mice is not affected by poly (I: C), which indicates that poly (I: C) mainly damages ovarian function by TNF- 伪 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.
【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R392
本文编号：2409902
[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- 伪, IL-6,IFN- 伪 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- 伪-/-mice is not affected by poly (I: C), which indicates that poly (I: C) mainly damages ovarian function by TNF- 伪 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.
【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R392
【参考文献】
相关期刊论文 前1条
1 ;An update on primary ovarian insufficiency[J];Science China(Life Sciences);2012年08期
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