阿加曲班改性滤器膜通过凝血酶介导的补体C5a/C5aR通路影响微炎症形成的机制研究

Hemodialysis is an important treatment for acute and chronic renal failure. However, the microinflammation caused by prolonged dialysis often leads to the formation of complications. Based on the recent development of complement C5a/C5aR pathway, thrombin and microinflammation, and the relationship of AG and thrombin, we proposed that " the modified hemodialysis filter membrane of agargyan can inhibit the thrombin to reduce the expression of CD88, and then the activation of neutrophil-activated cells are inhibited by inhibiting the C5a/CD88 pathway. On the other hand, it can inhibit the C5a/p38MAPK pathway to promote neutrophil autophagy and apoptosis to alleviate the state of micro-inflammation during dialysis ". On the basis of argatroban modified filter membrane (AG filter) constructed in the earlier stage, we will choose sheep and human as study object, thrombin (activating complement C5a), C5a blocker and other interventions were used, through the complement-neutrophil feedback mechanism, using ELISA, FCM, WB and other methods to explore the effects of ordinary dialysis membrane and AG membrane on microinflammatory markers (CRP, IL-6, TNF-α, etc.), and analyze C5a/C5aR pathway in the formation of micro-inflammatory effect. This project will clarify the formation of hemodialysis-related micro-inflammation and the anti-inflammatory mechanism of AG filter from a new perspective and provide the basis for promoting the clinical transformation of AG filter.

血液透析是急、慢性肾功能不全的重要治疗手段，但透析膜本身导致的微炎症状态会导致诸多并发症形成。基于补体C5a/C5aR（CD88）通路及凝血酶与微炎症形成的最新研究进展，以及阿加曲班与凝血酶的相互作用，我们提出以下假说：阿加曲班改性滤器膜（AG滤器膜）通过抑制凝血酶，下调C5a的表达，进而抑制C5a/CD88通路来抑制中性粒细胞激活；同时也通过抑制C5aR/p38MAPK通路促进中性粒细胞自噬和凋亡，从而改善透析过程中微炎症状态。本项目在前期构建的AG滤器膜基础上，以人体和绵羊为研究对象，采用凝血酶（激活补体C5a），C5a阻断剂等干预，通过补体-中性粒细胞反馈机制，运用ELISA、FCM、WB等方法，探索AG膜对微炎症指标的影响，解析凝血酶-C5a/C5aR通路在微炎症形成中的作用。本项目将从新的视角阐明血液透析微炎症形成及AG滤器抗炎症机制，并为促进AG滤器临床转化提供依据。

血液净化是急、慢性肾功能不全的重要治疗手段，同时在危重症患者如新冠肺炎、重症胰腺炎等所致全身炎症因子风暴的治疗中应用广泛。血液净化膜属于体外材料，与血液接触后会引起膜表面血栓形成和微炎症状态产生，引起相关并发症。优化血液净化膜材料是解决本身问题的最佳处理方法，然而目前却鲜有一款能同时具有优良抗栓和抗炎双效作用的国产透析器面世，因此进行新型抗凝抗炎改性透析膜/滤器的自主研发迫在眉睫，且具有广阔的临床需求和社会经济效益。.本课题组发现，目前对于透析膜的改性主要集中在如何通过抑制凝血因子达到抗凝目的，少有直接通过抑制血小板抗栓的改性膜研究；另一方面，膜材料本身所致微炎症状态的机制研究亦相对缺乏。鉴于上述原因，我们在前期制备阿加曲班改性聚砜膜（AG膜）的成功经验基础上，首次用血小板PAR1受体抑制剂沃拉帕沙改性聚砜膜，也尝试用血小板P2Y12受体抑制剂替格瑞洛改性聚醚砜膜，并评估了改性膜的理化性能和抗血小板性能。同时我们也证实了普通聚砜膜材料可以通过补体C5a/C5aR通路调控微炎症因子的表达，而AG膜可以通过抑制上述通路下调CD88的表达，从而减轻微炎症状态。.后续我们期待能通过抗凝（阿加曲班）和抗血小板（沃拉帕沙或替格瑞洛）两方面协同加强透析膜/血界面抗栓和抗炎性能，达到更高效的抗生物材料相关血栓及微炎症形成的目的，以期在危重症患者中开拓一种全新的血液净化理念与模式。