NUP98在病毒性心肌炎Th17细胞分化中的作用和机制研究

Although Th17 cells were recently reported to be involved in the development of viral myocarditis (VMC), the mechanism of Th17 cell differentiation induced by virus infection directly remains unknown. Nuclear pore complex proteins (Nups), the main components of the nuclear pore complex, mediate molecule transport between nucleus and cytoplasm, which regulates cell differentiations. Nup98, an important member of Nups, has been proved to be associated with virus replication and antiviral responses. Our group recently found that the levels of Nup98 were negatively correlated with RORγt that was the key transcription factor in Th17 cell differentiation in patients with VMC. Therefore, we hypothesized that Nup98 might be associated with virus-induced Th17 cell differentiation in VMC. In the present study, after establishing VMC mouse model by the common virus coxsackievirus B3 (CVB3), the expression of Nup98 would be regulated by transfection techniques, and the levels of Nup98 in Th cells, viral replication, and Th17 cell differentiation would be detected. Then the expressions of RORγt, STAT3, CBP/p300 and the binding ability among these transcription factors would be further explored. These new findings might clarify the role and mechanism of Nup98 in Th17 differentiation, which would reveal the new immunological pathogenesis of VMC.

病毒性心肌炎（VMC）的发生发展与Th17细胞密切相关，但是病毒感染后直接促进Th17细胞分化的机制至今尚未阐明。核孔蛋白（Nups）是细胞核孔复合体的主要组成部分，其参与调节细胞核与细胞质间分子的运输，从而介导细胞分化。Nup98是一种与病毒感染和机体抗病毒免疫密切相关的核孔蛋白。本课题组新近研究发现Nup98水平与VMC患者Th17细胞分化关键转录因子RORγt水平呈负相关。因此，我们推测Nup98可能参与VMC中病毒诱导的Th17细胞分化。本研究拟采用VMC中最常见的病原体柯萨奇病毒B3（CVB3）建立VMC小鼠模型，在体内外分别利用转染技术促进或抑制Nup98表达，检测Th细胞中Nup98水平、病毒复制、Th17细胞分化以及RORγt、STAT3、CBP/p300等转录因子的表达和结合能力等，探讨Nup98在VMC中Th17细胞分化的作用和机制，从而揭示VMC新的免疫学发病机制。

病毒性心肌炎（VMC）的发生发展与Th17细胞密切相关，核孔蛋白（Nups）是细胞核孔复合体的主要组成部分，其参与调节细胞核与细胞质间分子的运输。Nup98是一种与病毒感染和机体抗病毒免疫密切相关的核孔蛋白，本课题组以往研究发现Nup98水平与VMC患者Th17细胞分化关键转录因子RORγt水平呈负相关。因此，我们推测Nup98可能参与VMC中病毒诱导的Th17细胞分化。本研究采用VMC中最常见的病原体柯萨奇病毒B3（CVB3）建立VMC小鼠模型，在体内外分别利用转染技术促进或抑制Nup98表达，检测Th细胞中Nup98水平、病毒复制、Th17细胞分化以及RORγt、STAT3、CBP/p300等转录因子的表达和结合能力等。结果发现CVB3能够直接进入小鼠CD4+Th细胞，并能增加RORγt、IL-17合成，减少Nup98产生，从而促进CD4+Th细胞向Th17细胞分化。进一步研究中，我们在体内外分别利用pcDNA3.1-Nup98和siRNA-Nup98上调和下调Nup98产生，结果显示Nup98水平下降后，CBP/p300解离增多，STAT3乙酰化水平升高，RORγt合成增多，最终导致Th17细胞水平明显增加。Nup98水平升高后CBP/p300解离减少，STAT3乙酰化水平下降，RORγt合成减少，最终导致Th17细胞水平明显减低。我们的研究成果证实了Nup98在VMC中CVB3直接促进Th17细胞分化的作用和机制，从而揭示VMC新的免疫学发病机制。