基于细胞凋亡和B细胞稳态研究活血解毒方对干燥综合征的干预机制

Sjogren's syndrome（SS）could be triggered by some potential stimulation factors，who activating Caspase-1 by inflammasome，and produce mature IL-1βand IL-18. Exocrine glands local continuous state of micro inflammation promote lymphocyte proliferation，which may induce excessive B cell activation and target organ cell apoptosis increase，then exocrine gland atrophy and lead to the onset of SS. Cell apoptosis and the excessive activation of B cells are the important intermediate process. Preliminary work showed SS exhibit dynamic pathological changes: the dryness lead to blood stasis- excessive dryness turn to toxin- conglutination of dryness, blood stasis and toxin in SS patients. Huoxue Jiedu Formula could relieve the inflammation of salivary gland in NOD mice, improve the saliva secretion, reduce damage of target tissue and maintain B cells homeostasis. Activating blood circulation and removing blood stasis intervenes apoptosis. In this study, these works will be done as followed：（1）duplicating induced SS mouse model and treated with Huoxue Jiedu Formula，tissue morphology and molecular biology technique will be used to study the intervention mechanism of apoptosis, state of micro inflammation and regulation of B cells homeostasis.（2）Human submandibular gland cells will be used in vitro after induced apoptosis to study the mechanism of apoptosis intervened by the drug-containing serum. Purpose to study the intervention mechanism of Huoxue Jiedu Formula to Sjögren's syndrome on the basis of apoptosis and B cells homeostasis.

干燥综合征（SS）可能因某种潜在的刺激因素，通过炎症小体激活Caspase-1，产生成熟的IL-1β和IL-18，外分泌腺局部持续的微炎症状态导致淋巴细胞浸润，B细胞过度活化及靶器官细胞凋亡增加，腺体萎缩，导致SS发病。细胞凋亡和B细胞的过度活化是其中的重要中间过程。前期研究SS存在因燥致瘀-燥盛成毒-燥毒瘀胶着内伏的病变过程，活血解毒方可减轻NOD小鼠颌下腺炎症反应，改善唾液分泌量，减轻功能受损，维持B细胞稳态。活血化瘀具有调控细胞凋亡的作用。基于前期工作开展以下研究：（1）在体研究：复制诱导性SS小鼠模型，以活血解毒方为干预手段，从组织形态学、分子生物学等研究其调控细胞凋亡、微炎症状态，维持B细胞稳态的机制；（2）体外研究：以含药血清干预诱导凋亡的人下颌下腺细胞，从细胞层面研究活血解毒方对细胞凋亡调控机制。目的：从细胞凋亡和维持B细胞稳态的角度，研究活血解毒方对SS的干预机制。

干燥综合征（Sjögren's syndrome，SS）是一种以唾液腺和泪腺等外分泌腺体炎症反应和功能紊乱为特征，以口干、眼干等为主要表现，逐渐累及肌肉骨骼及消化、呼吸等多系统的慢性系统性自身免疫性疾病。近年研究表明，外分泌腺细胞的凋亡是本病早期病理变化之一。外分泌腺局部持续的微炎症状态导致淋巴细胞浸润，B细胞过度活化及靶器官细胞凋亡增加，腺体萎缩。干预靶器官的细胞凋亡、抑制B细胞的过度活化或许能为SS的治疗提供新思路。基于前期工作，从动物实验和细胞实验，以活血解毒方为干预手段，从组织形态学、分子生物学等研究其调控细胞凋亡、微炎症状态，维持B细胞稳态的机制。主要研究结果表明：1.固有免疫参与的细胞凋亡（内源性凋亡因子、Bcl-2基因家族、外源性途径凋亡因子）、获得性免疫参与的淋巴细胞浸润、B细胞的过度活化以及相关信号通路在SS中存在着动态变化；2.B细胞过度活化可以通过IFN/BAFF/APRIL轴、IL-6/IL-21轴进行调控；3.活血解毒方可能是通过调控干燥综合征细胞凋亡（内源性凋亡因子、Bcl-2基因家族、外源性途径凋亡因子）以及微炎症状态，维持B细胞稳态的生物学机制来实现的，完善了该方治疗SS的作用机制，进一步揭示了活血解毒、滋阴润燥的科学性。