miR-30调控Th17分化在多发性硬化发病中的作用和机制

The excessive differentiation of T helper cell 17 (Th17) can cause multiple sclerosis (MS). MicroRNA (miR) is able to regulate Th17 differentiation by silencing target gene. Our previous data showed that the expression of miR-30 decreased gradually during the differentiation of naive T cells (Th0) to Th17; while the percentage of Th17 increased, the miR-30 level was significantly reduced in the lesion area of MS mice. Thus, we speculate that miR-30 could block Th17 differentiation through silence matching target gene to inhibit the pathogenesis of MS. To screen the target gene of miR-30 and confirm our hypothesis, firstly, we will detect the rate of Th17 differentiation by using miR-30 over-expressed Th0. Secondly, in the miR-30 over-expressed mouse model, we will study the role of miR-30 on Th17 differentiation and the pathogenesis of MS in vivo. Thirdly, we will screen the target gene of miR-30 and investigate the mechanism underlying the regulatory effect of miR-30 on Th17 differentiation using bioinformatics software, gene expression chip, dual luciferase reporting system, and RNA interference technology. Finally, we will investigate the effect of miR-30 on the Th17 differentiation and pathogenesis of MS in target gene defect mice to further confirm the regulatory mechanism of miR-30 in vivo. This work investigating the role and mechanisms of miR-30 regulated Th17 cell differentiation in the pathogenesis of MS will provide important theoretical basis for the research and treatment of MS.

辅助性T细胞17（Th17）的过度分化可诱发多发性硬化（MS）。微小RNA（miR）能够介导相应靶基因的沉默而调控Th17分化。我们前期研究发现：幼稚T细胞（Th0）分化为Th17过程中，miR-30表达逐渐下降；随MS小鼠病变区Th17比例上升，miR-30含量明显减少。故推测：miR-30可介导相应靶基因的沉默而阻断Th17分化，进而抑制MS发病。为筛选miR-30的靶基因并证实我们的推测，本项目拟在Th0中过表达miR-30，检测Th17的分化；小鼠过表达miR-30，观察其对Th17分化及MS发病的影响；用生物信息学、基因芯片、双荧光素酶系统和RNA干扰技术筛选miR-30调节Th17分化的靶基因；靶基因缺失小鼠过表达miR-30，观察Th17分化及MS发病情况。本项目可揭示miR-30调控Th17分化在MS发病中的作用和机制，为MS的基础研究和临床治疗提供理论依据。

辅助性T细胞17（Th17）的过度分化可诱发多发性硬化（MS）。微小RNA（miR）能够介导相应靶基因的沉默而调控Th17分化。我们前期研究发现：幼稚T细胞（Th0）分化为Th17过程中，miR-30表达逐渐下降；随MS小鼠病变区Th17比例上升，miR-30含量明显减少。故推测：miR-30可介导相应靶基因的沉默而阻断Th17分化，进而抑制MS发病。为筛选miR-30的靶基因并证实我们的推测，本项目在Th0中过表达miR-30，检测Th17的分化；小鼠过表达miR-30，观察其对Th17分化及MS发病的影响；用生物信息学、基因芯片、双荧光素酶系统和RNA干扰技术筛选miR-30调节Th17分化的靶基因。研究结果显示在Th0中过表达miR-30a后抑制Th17生成，降低RORγt、IL-17A和IL-17F的表达。EAE小鼠过表达miR-30a后症状明显减轻，表现为kono评分降低，脱髓鞘程度较轻，髓鞘结构相对完好，MBP的表达升高，炎性细胞浸润较少，淋巴结、脾脏及外周血中CD4+IL-17+T细胞比例下降，同时IL-17水平降低。miR-30a的种子序列可以与Th17分化相关的IL-21R的3’UTR互补配对，且双荧光素酶报告系统显示其是miR-30a的直接靶基因。此外，临床MS患者体内的miR-30a表达与Th17的分化呈现一定的负相关，也进一步证实了miR-30a对Th17的抑制作用。本项目揭示miR-30调控Th17分化在MS发病中的作用和机制，为MS的基础研究和临床治疗提供理论依据。