芪丹汤基于肥大细胞TNF-α调控腹膜间皮细胞NLRP3/IL-1β抗腹膜纤维化的机制研究

腹膜纤维化(PF)是腹透患者超滤衰竭的主要原因。肥大细胞参与PF，但机制不明。脂多糖(LPS)可使肥大细胞活化，后者释放的肿瘤坏死因子-α(TNF-α)等可激活腹膜间皮细胞(PMCs)NLRP3/IL-1β，介导炎症和转分化。预实验表明，以补气活血为主要治法的芪丹汤（黄芪、丹参）可抑制PMCs转分化。我们提出芪丹汤基于肥大细胞TNF-α调控PMCs NLRP3/IL-1β干预腹膜纤维化这一假说。本研究拟建立肥大细胞与PMCs共培养体系，观察LPS激活的肥大细胞TNF-α释放，及后者对PMCs NLRP3/IL-1β及转分化指标的影响。同时，构建腹透小鼠模型，观察芪丹汤、TNF-α抑制剂及肥大细胞稳定剂作用下PMCs NLRP3/IL-1β及转分化指标。本研究旨在探讨肥大细胞在PF中的作用，研究芪丹汤干预PF的机制，为中药治疗PF提供新的靶点，完善补气活血的中医治法理论在PF中的应用。

Peritoneal fibrosis (PF) is the leading cause of ultrafiltration failure and discontinuation of peritoneal dialysis. It is known that the number of mast cells is significantly increased in the fibrotic peritoneum. However, the precise mechanism of mast cells in PF is still unknown. Mast cell can be activated by Lipopolysaccharide (LPS). Tumor necrosis factor-α(TNF-α) can be released during mast cell degranulation. It is proved that TNF-α could activate NLRP3/IL-1β pathway, which has been demonstrated to contribute to peritoneal mesothelial cells (PMCs) mesothelial-mesenchymal transition (MMT). In our previous study, Qidan decoction, whose effect is to invigorating qi and activating blood circulation, is proved to ameliorate PF by inhibiting TGF-β. We speculate that Qidan decoction ameliorate PF by acting on NLRP3/IL-1β pathway in PMCs through TNF-α released by peritoneal mast cells. In this study, mast cells activated by LPS will be co-cultured with PMCs, and the Qidan decoction serum will be added to the culture dish. TNF-α, NLRP3/IL-1β, and markers of PMCs MMT will be detected. Meanwhile, intraperitoneal injection of peritoneal dialysis fluids (PDF) on mice will be done to establish PD mice model. Then, mice will be treated with Qidan decoction, mast cell stabilizer, or TNF-α inhibitor. Markers of NLRP3/IL-1β, and PMCs MMT will be detected. This study aims to explore the role of mast cells in PF, and mechanism of Qidan decoction on peritoneal mast cells. We aim to find a new target for the treatment of PF, and above all, to establish and complete the traditional Chinese medicine theory of invigorating qi and activating blood circulation.