基于B10细胞调控T细胞分化可塑性研究新风胶囊改善原发性干燥综合征 唾液分泌的机制

Primary Sjogren's syndrome (pSS) is an autoimmune disorder of salivary glands and other exocrine glands with associated lymphocytic infiltrates. The pathogenesis of pSS is considered to be related to the interaction of cellular immunity and humoral immunity caused excessive inflammation. Regulatory cells always play an important role to maintain the immune balance and homeostasis in body, and regulatory B cells (B10 cells) play a key role in immune regulatory for pSS. But like other autoimmune diseases, the specific regulation mechanism of pSS remains unclear. In our previous studies, we found that the elevated level of pro-inflammatory cytokine in supernatant of T cells culture, which remove B10 cells by MACS in pSS model rats. We speculate that B10 cells can negatively regulate T cells in pSS model rats. According to Traditional Chinese Medicine, using Chinese medicine compound Xinfeng capsule, "nourishing qi to invigorate spleen and dissipating dampness to dredge collaterals", which can effectively improve the salivary glands pathological damage and promoting salivation in pSS patients. Based on the previous clinical and experimental study, we propose the hypothesis that the Xinfeng capsule can intervene the process of B10 cells, which can regulate the differentiated plasticity of T cell subsets, improving the salivary glands pathological damage, restoring the Muc7, AQP5 expression and sensibility of M3R, and promoting salivation in pSS model rats. In vivo and in vitro, using MACS, CBA and cell adoptive transfer experiments, we study the mechanism that Xinfeng capsule regulate T cell differentiation plasticity through B10 cells, which promoting negative immune regulation, improving salivary glands pathological damage and eventually promoting salivation in pSS. Taken together, our research provides novel insights into the mechanism of pSS with Chinese medicine treatment.

原发性干燥综合征(pSS)是一种主要以淋巴细胞浸润累及唾液腺等外分泌腺的自身免疫性疾病。调节性细胞对免疫平衡和自身稳定的维持起重要作用，其中调节性B细胞（B10细胞）可负向调节T细胞亚群分化。前期研究发现，培养pSS模型大鼠去除B10细胞后的T细胞，培养上清促炎细胞因子水平显著升高。根据中医理论，采用“益气健脾通络”中药复方新风胶囊治疗pSS，能够有效改善患者唾液腺病理损伤，促进唾液分泌。综合前期实验和临床研究，课题组提出“采用新风胶囊干预B10细胞，调控T细胞亚群分化可塑性，降低唾液腺病理损害，恢复Muc7，AQP5表达及M3R敏感性，改善pSS唾液分泌”假说。采用体内外相结合的方法，运用磁珠分选、CBA流式液相芯片、细胞过继转移等技术，研究新风胶囊通过干预B10细胞，负性调控T细胞亚群分化和炎症反应，降低病理损害，最终改善pSS唾液分泌。为中医药改善pSS唾液分泌提供实验依据。

原发性干燥综合征(primary Sjogren's syndrome, pSS)是一种主要累及全身外分泌腺的慢性自身免疫性疾病，唾液腺是最常累及的腺体之一。pSS的发病机制尚不明确，可能与细胞免疫、体液免疫等因素的相互作用引起过度炎症反应及免疫损伤有关。调节性B细胞(B10细胞)在通过其分泌的IL-10参与自身免疫性疾病负性免疫调节过程中发挥重要的作用。. 新风胶囊是经安徽省食品药品监督管理局批准注册的院内制剂(皖药制字Z20050062)。临床研究发现新风胶囊能明显提高pSS患者唾液流率，改善患者口干症状。本课题以“脾虚致痹，从脾论治”理论为指导，通过建立pSS动物模型，B10细胞为主线，唾液分泌功能为靶点，从免疫调节角度探讨新风胶囊改善pSS唾液分泌机制。. 研究表明，新风胶囊可显著改善pSS模型小鼠唾液分泌。模型鼠颌下腺组织病理学观察与评分，相对于模型组，新风胶囊干预组小鼠颌下腺组织炎细胞浸润显著恢复，流式细胞学同时发现，颈部淋巴结B10细胞亚群比例显著提升，颌下腺水通道蛋白AQP5表达上调。通过对颌下腺组织测序及后续验证，课题组发现pSS致病过程中关键基因EGR1。通过唾液蛋白组学筛选发现，C3、CFH、SERPING1、FGA和FGG唾液蛋白水平与pSS疾病进展显著相关，可作为pSS诊疗潜在的生物标志物和干预靶点。. 本课题的顺利实施将有助于从免疫学、唾液蛋白组学角度阐明pSS的发病机制，明确新风胶囊改善pSS唾液分泌的作用机制，为pSS的诊疗提供新的思路，亦为中医药干预pSS进行了有益的探索。