miR-142-3p/5p靶向CLIC4调控弓形虫感染过程中宿主细胞凋亡的机制研究

弓形虫严重威胁人类和动物健康，目前尚无理想的治疗药物。我们的前期研究发现宿主miR-142-3p/5p在弓形虫感染中显著上调表达，而CLIC4 mRNA和蛋白下调表达。miR-142-3p/5p和CLIC4广泛参与了细胞凋亡。经生物信息学分析，CLIC4是miR-142-3p/5p的潜在靶基因。因此我们推测miR-142-3p/5p靶向CLIC4调控弓形虫感染过程中宿主细胞凋亡。为此，我们拟首先利用双荧光素酶报告系统等对miR-142-3p/5p与CLIC4的靶向关系进行验证；接下来过表达或敲低miR-142-3p/5p或CLIC4，检测感染弓形虫后宿主细胞凋亡率及凋亡相关蛋白;最后利用基因敲除小鼠模型研究miR-142基因是否影响宿主对弓形虫的易感性。本研究将深入解析弓形虫感染的致病机制，并为找到防治弓形虫病的新靶点奠定科学依据。

Toxoplasma gondii is a serious hazard to both human and animal health. However, ideal drugs are not yet available.Our previous studies found that miR-142-3p/5p were up-regulated during T. gondii infection. Meanwhile, the expression levels of chloride intracellular channel 4 (CLIC4) mRNA and protein were decreased.Using literature search and online bioinformatics software, we found CLIC4 was a potential target gene regulated by both miR-142-3p and miR-142-5p. Therefore, we postulated that host apoptosis was regulated by the manipulation of host miR-142-3p/5p/CLIC4 axis during T. gondii infection. To verify the hypothesis, we will firstly confirm whether miR-142-3p/5p targets CLIC4 by using dual-luciferase reporter gene assay. Molecular that are involved in apoptosis and apoptosis rate of host cells will be detected following over/down-expression of miR-142-3p/5p or CLIC4 with T. gondii infection. Finally, to explore whether miR-142 influences host resistance during infection, the survival rate of miR-142 knock-out mice will be recorded after virulent or avirulent parasite infection. This study will not only shed lights for further studies of the pathogenesis of T. gondii infection, but also provide new theoretical supports for the exploration of novel therapeutic targets against toxoplasmosis.