解毒活血法调控LC3相关吞噬（LAP）抑制狼疮鼠自身炎症反应的作用和机制

Defects in apoptotic cell clearance have been linked with auto-inflammatory response in systemic lupus erythematosus (SLE). New findings show that deficiencies in a form of noncanonical autophagy pathway, known as LC3-associated phagocytosis (LAP), result in defective digestion of engulfed dying cells, which is sufficient to break immune tolerance, trigger inflammatory cytokine secretion and exacerbate renal injury in SLE. Our previous studies showed that Jiedu Huoxue Treatment Method, represented by Lupus Recipe, can effectively ameliorate the inflammatory conditions and renal injury in SLE with noxious heat and blood stasis syndrome. Proper autophagy could protect cells from apoptosis. However, whether Jiedu Huoxue Treatment Method inhibits the inflammatory response in SLE by re-elicit the noncanonical autophagy pathway remains unclear. Here, we hypothesize that this therapy may have an anti-inflammatory effect on SLE by modulation of LAP pathway to engulf and digest dying cells, thus it attenuate the auto-inflammatory response, autoantibody production, and target organ injuries. In this study, we are going to investigate the role of Jiedu Huoxue Treatment in regulating macrophage LAP and its mechanism for attenuating the auto-inflammation and renal injury in SLE with spontaneous lupus mice model, gene knockout, immunopathology method, etc. Furthermore, the dynamics between Jiedu Huoxue Treatment and corticosteroids will also be studied based on clinical model. This study will reveal the modern scientific connotation of Jiedu Huoxue Treatment and provide novel experimental evidence for its clinical application in SLE therapy.

凋亡细胞清除障碍与系统性红斑狼疮（SLE）自身炎症反应相关。吞噬细胞非经典自噬通路—LC3相关吞噬（LAP）缺陷不能有效降解吞噬的濒死细胞，导致SLE免疫耐受破坏、炎症细胞因子分泌及肾组织损伤。我们以往研究发现以狼疮方为代表的解毒活血法可有效改善SLE热毒血瘀证炎症状态和肾损伤；适度自噬可保护细胞免于凋亡。然而解毒活血法是否通过调控非经典自噬抑制SLE自身炎症反应尚不清楚。我们假设解毒活血法的抗炎作用可能通过调控巨噬细胞LAP通路吞噬和降解濒死细胞，抑制SLE自身炎症反应、自身抗体产生及靶器官损伤。本研究拟利用自发性狼疮鼠模型、基因敲除及免疫病理等技术，探讨解毒活血法对狼疮鼠巨噬细胞LAP通路的调控作用及其改善SLE炎症状态和肾损伤的机制；并基于临床实践模式探讨解毒活血法与糖皮质激素联合应用对LAP通路的协同调节作用，为SLE中医解毒活血法现代科学内涵的揭示及其临床应用提供新的实验依据。

本项目已完成研究。我们的研究结果发现：① 狼疮方能抑制MRL/LPR狼疮模型小鼠的自身免疫炎症反应，抑制肾脏细胞凋亡，改善狼疮小鼠肾脏损伤。② 尾静脉注射DBA/2脾脏和胸腺混合细胞悬液可在B6D2F1小鼠中成功建立狼疮模型。狼疮方还可缓解cGVHD狼疮模型小鼠的病变进展，保护肾功能，减轻肾脏损伤。③ 狼疮方可抑制cGVHD狼疮模型小鼠肾功能下降，减轻肾脏损伤。狼疮方可提高cGVHD狼疮模型小鼠非经典自噬蛋白的表达水平，而对经典自噬蛋白的影响差异不显著。狼疮方与激素联合用药能更显著地提高狼疮小鼠肾脏非经典自噬蛋白的表达水平。狼疮方与激素可能协同作用于非经典自噬通路抑制cGVHD狼疮模型小鼠的肾脏损伤。综上所述，本研究解毒活血法调控LC3相关吞噬（LAP）抑制狼疮鼠自身炎症反应的作用和机制，揭示了中药复方狼疮方通过调控非经典自噬通路抑制狼疮小鼠自身免疫炎症反应以及肾脏损伤的可能机制。