卵转铁蛋白增强巨噬细胞抗病毒天然免疫作用的分子机制

Ovotransferrin(OVT) is one of the major bioactive compounds in egg, and has good antiviral activities. Our previous research work demonstrated that OVT could significantly inhibit the vesicular stomatitis virus (VSV) replication on mouse peritoneal macrophages, but could not affect the binding of the virus on macrophages. Is the antiviral function of OVT achieved by the way of stimulating gene expression and then elevating the antiviral cytokine levels, or via any other mechanisms? All these questions need to be addressed. In this study, qRT-PCR, ELISA, Western Blot, and other comtemperary and conventional methodologies will be used to determine the effects of OVT on the secretion of type I interferon and related proteins expressions of macrophages induced by VSV in vitro. Meanwhile, the effects of OVT on the nuclear transfer factor activation in macrophages induced by TLRs and TfR will be studied to analyze the correlationship between the signal pathway mediated by TLR or RIG-I and INF-1 expression, and thus to elucidate the molecular mechanisms of the OVT enhancing macrophages antiviral innate immunity at the cellular and molecular levels. The expected results will provide a theory basis for OVT being used as a functional factor in health food producion, which will have profound impacts and academic values for the further utilization of the OVT.

卵转铁蛋白（OVT）是禽蛋中主要活性蛋白之一，具有较好的抗病毒作用。我们前期研究表明OVT不影响水泡口炎病毒（VSV）与巨噬细胞的结合，但能显著地抑制VSV在小鼠腹腔巨噬细胞内的复制。OVT这种抗病毒作用是通过何种机制来实现的？目前尚未见相关报道。本研究拟采用体外细胞培养模型，利用实时荧光定量RT-PCR、ELISA和Western Blot 等技术分析OVT对VSV诱导巨噬细胞干扰素基因转录、干扰素分泌及相关蛋白的表达的影响；研究OVT对巨噬细胞经TLRs、TfR诱导的核转入因子活化的影响，及活化的转入因子与TLRs 或RIG-I介导的INF-I表达的相关性。从细胞和分子水平阐明OVT增强巨噬细胞抗病毒天然免疫作用的分子机制。本研究结果可为卵转铁蛋白作为食品功能因子在保健食品中的应用提供理论依据，对禽蛋的深度开发利用具有重要的影响。

卵转铁蛋白（ovotransferrin，OVT）是禽蛋蛋清的主要组成蛋白。大量研究证明OVT具有多种免疫活性及抗病毒免疫活性，然而对OVT抗病毒的机理的研究并不是十分清楚。因此，本研究采用体外细胞培养的模型，从细胞和分子水平阐明OVT抗病毒天然免疫作用的分子机制。. 结果表明：OVT孵育的小鼠巨噬细胞，经RNA病毒感染后，显著抑制了病毒的复制；OVT通过抑制开放基因表达的H3K4和促进抑制基因表达的H3K27的三甲基化，从而抑制细胞内RNF125分子的表达，而RNF125负调控RIG-I分子，其表达下降促进RIG-I蛋白的分泌；OVT是通过通过RIG-I/IRF-3通路诱导I型干扰素分泌的上调，从而发挥抗病毒作用。. 该研究为卵转铁蛋白及其酶解物的应用提供理论依据，开发禽蛋深加工与利用的新方向，同时也为活性蛋白增强机体抗病毒天然免疫的研究提供研究基础。