本课题组以前期申请的项目（81271759）为依托，发现SLE患者血清HIF-1α表达升高，且表达水平与SLE 疾病活动度和血清IL-17 水平呈正相关，提示异常表达的HIF-1α参与了SLE的发病。值得注意的是，已有证据表明ROS可诱导HIF-1α 的表达，提示ROS 可经过HIF-1α 影响Th17/Treg 平衡，进而导致炎性因子的表达紊乱。而课题组前期与英国萨里大学Wu Changhao教授合作研究发现SLE患者T 细胞内ROS 表达显著升高。因此，上述结果为探讨机体氧化应激状态在SLE 病理机制中的作用提供了重要依据。鉴于此，本项目拟在前期基金研究基础上，借助Wu教授课题组在疾病发病中的生理及病理机制研究经验（特别是ROS 的测量、siRNA 技术及基因敲除小鼠应用领域），以及最近Wu教授ROS 课题获得NIH 五年资助的有利条件，进一步研究HIF-1α 上游调控因子ROS 在SLE发病中的初步功能，以增加原课题的深度。同时，通过与英方合作与交流，深入开展依托项目计划的基于狼疮小鼠的动物实验，初步阐明HIF-1α 在SLE 发病中的分子机制，为发现新的治疗靶点提供科学依据。

With the support of the National Natural Science Foundation of China (81271759), our research group had finding that serum levels of HIF-1α in patients with systemic lupus erythematosus (SLE) were significantly increased and positively associated with disease activity index and serum levels of IL-17. The findings indicated that abnormal expression of HIF-1α participates in the pathogenesis of SLE. Notably, available evidence showed that ROS can enhance HIF-1α expression, which suggesting that ROS can result in disordered proinflammatory cytokines expression by influencing imbalance of Th17/Tregs through HIF-1α. Furthermore, our group together with Professor Wu Changhao in University of Surrey had also demonstrated that ROS expression in T cell was increased in SLE patients than in controls. The above results thus suggest that oxidative stress could also plays an important role in pathogenesis of SLE. Therefore, with the support of experience from Professor Wu’s research group in the physiological and pathological mechanisms (especially ROS detection, siRNA, gene knockout mice) and recently advantageous condition that Professor Wu’s ROS research obtaining five-years fund from NIH, we intend to study the initial function of upstream regulatory factor (ROS) in SLE so as to increase the profundity of the previous research. Meanwhile, by the cooperation and exchange with Wu’s group, we hope to make greater progress on previous foundation program (animal experiment in lupus mice). The final findings will help us to clarify the molecular mechanisms of HIF-1α deficiency in the development of SLE and thus provide a scientific basis for new therapeutic targets.

本课题组以前期申请的国家自然基金面上项目（81271759）为依托，发现系统性红斑狼疮（SLE）患者血清低氧诱导因子1α（HIF-1α）表达异常，且表达水平与SLE 疾病活动度评分和血清IL-17 水平呈正相关，提示异常表达的HIF-1α参与了SLE的发病。而课题组前期与英国萨里大学Wu Changhao教授课题组合作研究发现SLE患者T 细胞内ROS 表达显著高于正常对照。值得注意的是，已有证据表明ROS可诱导HIF-1α 的表达，提示ROS 可经过HIF-1α 影响Th17/Treg 平衡，进而导致炎性细胞因子的表达紊乱。鉴于此，本项目拟在前期国家基金的研究基础上，借助Wu教授课题组在疾病发病中的生理及病理机制研究经验（特别是Ros检测、siRNA 技术及基因敲除小鼠应用等领域），深入开展依托基金项目计划的基于狼疮小鼠的动物实验，从整体上观察ROS下游因子HIF-1α 缺陷对狼疮小鼠疾病发展的影响，初步阐明其在SLE 发病中的分子机制，为发现新的治疗靶点提供科学依据。通过互访交流，我们完成了动物实验，结果发现：与对照小鼠相比，干扰后2周时HIF1α-shRNA狼疮小鼠血清IL-17表达下降，差异具有统计学意义；与对照组相比，干扰后4周时血清ANA下降，差异具有统计学意义； HIF1α-shRNA组干扰后24 小时尿蛋白与干扰前相比下降，干扰前后差值与对照组相比，差异有统计学意义；与对照组相比，HIF1α-shRNA组三个时间点病理损伤均有所减轻，但仅4 w时达到统计学差异；与对照组相比，各时间点HIF1α-shRNA组IgG、C3沉积均有下降。综上，我们的动物实验结果进一步揭示了HIF1α缺陷导致SLE发生发展的分子机制，并为采取新的治疗措施提供了科学依据。