TSH激活GRK2促进单核细胞迁移在动脉粥样硬化形成中的机制研究

单核细胞迁移是动脉粥样硬化形成的重要环节，而G蛋白偶联受体激酶2(GRK2)磷酸化在单核细胞迁移过程中发挥重要作用。我们前期研究表明促甲状腺激素（TSH）促进动脉粥样硬化的形成，同时发现TSH/TSHR与单核细胞的迁移有关。体外si-RNA敲减GRK2后TSH诱导的单核细胞迁移作用减弱。因此，本项目假设：TSH可能通过GRK2调控单核细胞迁移，促进动脉粥样硬化的形成。本项目在动物水平，利用单核细胞特异性敲除TSHR及GRK2的两种基因敲除小鼠模型，亚甲减及动脉粥样硬化两种疾病模型，应用单核细胞过继转移、荧光微球追踪等方法，论证课题假设。在细胞水平，利用原代单核细胞及细胞系，通过干预GRK2及其相关通路分子、筛选GRK2蛋白磷酸化位点等手段，明确TSH激活GRK2的信号分子及TSH活化GRK2磷酸化位点。本项目将揭示TSH在单核细胞迁移中的作用机制，为防治动脉粥样硬化提供新的思路和靶点。

Monocyte migration is an important part of the development of atherosclerosis, and G protein coupled receptor kinase 2 (GRK2) phosphorylation plays an important role in monocyte migration. Our previous studies have shown that thyroid stimulating hormone (TSH) promoted the formation and development of atherosclerosis, and TSH/TSHR was related to monocyte migration. After GRK2 was knockdown by si-RNA, the migration of monocytes induced by TSH was decreased obviously. Therefore, this project hypothesized that TSH activated GRK2 may regulate monocyte migration and promote the development of atherosclerosis. In this project, we introduced monocyte specific knockout TSHR or GRK2 mice models, and induced subclinical hypothyroidism or atherosclerosis disease models. In vivo, we used the methods of monocyte adoptive transfer and fluorescent microsphere tracking to demonstrate the hypothesis of the project. In vitro, we cultured primary monocytes and cell lines and identified the signal molecules and phosphorylation sites of GRK2, by intervening pathway molecules and screening GRK2 protein phosphorylation sites. This project will reveal the mechanism of TSH in monocyte migration and provide a new target for the prevention and treatment of atherosclerosis.