益气活血化湿法干预IMN发病中Th17/Treg免疫失衡和足细胞免疫损伤的调控机制及拆方研究

The idiopathic membranous nephropathy (IMN) is the most common cause of nephrotic syndrome in adults. As the primary approach for this disease, immunodepression still remains controversy due to little effectiveness as well as the serious side effect. Contemporarily, it is believed that this condition is autoimmune kidney disease induced by antibody provoked by antigen which located on the surface of the podocyte. The activation of complement and impairment of podocyte induced by the in situ immune complex under the epithelium is the cause of the proteinuria..The immune inbalance of Th17/Treg plays an important role in the development of IMN. podocyte is not only the target of the antigen but also the main etiological factor during the pathogenesis of the disease..TCM therapy has been proved to be effective for this disease, and the mechanisms behind its biological activity will be revealed in the present study from a point of view of immunity regulation. Firstly, we intend to elucidate the immunity modulation of Th17/Treg immune imbalance of Chinese herbs. These herbs are categorized by the different treatment principles including qi nourishing, invigorating the circulation of blood as well as resolving dampness. Each of the above treatment principles will be observed separative and cooperatively on the rat MN model. Secondly, we will discover the mechanism of these herbs on prevention of podocyte immunologicmjury. Finally, we intend to investigate the value of the above treatment principles in maintaining the normal function of the kidney-immuno-microenvironment..We are striving to discover the immunity modulation and effector targets of the above treatment principles respectively, in order to optimizing the formula and so as to setting a solid foundation for further drug development.

特发性膜性肾病（IMN）是最为常见的导致成人肾病综合征的原发性肾小球疾病，免疫抑制治疗疗效不理想且毒副作用严重。目前认为IMN是由抗足细胞表面抗原成分的抗体介导的自身免疫性肾脏病，其上皮下原位免疫复合物形成导致的补体活化以及足细胞损伤是IMN大量蛋白尿产生的原因。Th17/Treg免疫失衡在IMN发病中具有重要作用；足细胞既是IMN中免疫损伤的靶细胞，同时又主动参与到免疫发病机制中。在前期取得明确临床疗效的基础上，本研究拟从IMN发病的免疫机制入手，一方面探讨"益气活血化湿"中药及其拆方对MN大鼠模型Th17/Treg免疫失衡的调控机制；一方面着眼于IMN中足细胞的免疫损伤机制，探讨本法中药的足细胞保护机制，探索该法改善肾脏局部免疫微环境、保护足细胞的作用机制。力图揭示"益气"、"活血"、"化湿"各法在免疫调控及足细胞保护中的作用靶点与机制，从而优化组方为进一步开发新药打下坚实基础。

特发性膜性肾病（IMN）是最为常见的导致成人肾病综合征的原发性肾小球疾病，免疫抑制治疗疗效不理想且毒副作用严重。目前认为IMN是由抗足细胞表面抗原成分的抗体介导的自身免疫性肾脏病，其上皮下原位免疫复合物形成导致的补体活化以及足细胞损伤是IMN大量蛋白尿产生的原因。Th17/Treg免疫失衡在IMN发病中具有重要作用；足细胞既是IMN中免疫损伤的靶细胞，同时又主动参与到免疫发病机制中。在前期取得明确临床疗效的基础上，本研究拟从IMN发病的免疫机制入手，一方面探讨“益气活血化湿”中药及其拆方对MN大鼠模型Th17/Treg免疫失衡的调控机制；一方面着眼于IMN中足细胞的免疫损伤机制。.　　本项目完成了稳定PHN大鼠模型的造模流程，为后续研究打下了基础。动物实验结果发现，造模第15天模型成功，环孢素组24小时尿蛋白定量较模型组显著下降，但中药组尚未表现出明显优势，中药组的作用体现在对血清总蛋白（TP）的提升及对肾功能的保护方面，这一现象与临床实际情况较为相符；采用流式细胞术动态观察PHN（被动Heymann肾炎）大鼠模型外周血中Treg细胞比例、表型与功能变化，研究结果发现，环孢素组在造模的第7、15及第22天均表现为CD4+、CD28+、Treg明显下降,有统计学差异（P<0.05)；而与环孢素组相比，在第1和15天中药各组CD4+、CD28+、Treg均明显升高，以全方组、益气组及活血组效果更为显著，提示中药各组对Th17/Treg免疫平衡具有显著的调解作用；各中药组对PHN大鼠肾组织及足细胞表面趋化因子CXCL16、CD2AP、FcRn等具有一定的调控作用。上述研究结果尚在进一步验证中。