干眼是常见高发的免疫性疾病，T细胞分化异常具有重要作用。研究显示细胞代谢重塑（EMR）调控T细胞分化，间充质干细胞（MSCs）调节T细胞EMR，胸腺素β4对MSCs有重要调控效应。前期工作原创性发现干眼存在眼表组织代谢重塑，重组人胸腺素β4（rhTβ4）联合MSCs显著提高干眼治疗效果。本项目创新性提出rhTβ4增强MSCs调节T细胞EMR治疗干眼的假说。采用高分辨率质谱组织成像、能量代谢分析等技术，利用干眼动物模型确证rhTβ4或/和MSCs的治疗效果，研究其纠正T细胞糖酵解代谢偏移、抑制T细胞过度分化及其诱导炎症反应的效应；体外实验解析rhTβ4影响MSCs增殖、迁移募集和调节免疫炎症的机制；共培养技术明确T细胞EMR对rhTβ4增强MSCs调控免疫炎症的关键作用；阐明T细胞EMR调控亚群分化和炎症因子的机制。丰富MSCs治疗干眼的理论内涵，提供rhTβ4联合MSCs治疗新方案。

Dry eye is a common immune inflammatory disease, in which the abnormality of T cell differentiation plays an important role. Studies have shown that the energy metabolism reprogramming (EMR) regulate T cell differentiation. Mesenchymal stem cells (MSCs) regulate the EMR of T cell and thymosin β4 have an important regulatory effect on MSCs. Our previous works found that there was metabolism reprogramming within ocular surface in dry eye and recombinant thymosin β4 (rhTβ4) combined with MSCs significantly improved the therapeutic effect of dry eye. This project innovatively proposes that rhTβ4 enhancing MSCs' regulation of T-cell EMR is the key mechanism of dry eye’s treatment. High resolution MALDI-TOF/TOF mass spectrometry imaging and energy metabolism analysis will be used to confirm the therapeutic effect of rhTβ4 or/and MSCs by using the animal model of dry eye; to study the effects of correction on T-cell glycolysis, the inhibition on T-cell over-differentiation and induced inflammatory response. We also use in vitro experiment to analyze the mechanism of rhTβ4 enhancing the proliferation, migration, recruitment and regulation of immune inflammation of MSCs. The key role of T cell EMR which rhTβ4 enhancing the regulation of immune inflammation of MSCs will be clarified by co-culture technology. The mechanism of T cell EMR regulating subpopulation differentiation and inflammatory factors will be elucidated. Our project will enrich the theoretical plan of MSCs into the treatment of dry eyes and provide a new clinical scheme of rhTβ4 combined with MSCs.