Department of Gastroenterology, The First Affiliated Hospital of Xiamen University, School of Medicine, Xiamen University, Xiamen, China.
Department of Gastroenterology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
J Biochem Mol Toxicol. 2023 Feb;37(2):e23242. doi: 10.1002/jbt.23242. Epub 2022 Oct 13.
The role of reactive oxygen species (ROS) is crucial for the pathogenesis of acute pancreatitis (AP). Proanthocyanidins (PAs) have been confirmed to exert antioxidant activity. Our study aimed to determine whether PAs alleviated SAP via reducing ROS, suppressing NLRP3 inflammasome, and inhibiting M1 macrophage polarization. Our study investigated the protective effects of PAs on pancreatic histopathological injury using SAP mice. The effects of PAs on macrophages were investigated in inflammatory RAW 264.7 cells or mouse bone marrow-derived macrophages (BMDMs) induced by lipopolysaccharide (LPS). Immunofluorescence staining and/or western blot assay were employed to evaluate NLRP3 inflammasome in macrophages and pancreatic tissue. Cell counting kit-8 (CCK-8) was used to access effects of PAs on cell viability and cytometry flow was used to determine the effects of the PAs on the ROS levels of the RAW 264.7 cells. Then, we evaluated M1 macrophage polarization using flow cytometry or real-time quantitative polymerase chain reaction (RT-qPCR). PAs administration alleviated pancreatic inflammation in SAP mice. The PAs depressed NLRP3 inflammasome and inhibited M1 macrophage polarization in pancreatic tissue. We also found that the PAs showed no cellular toxicity but decreased ROS levels in RAW 264.7 cells, downregulated the NLRP3 inflammasome in the macrophages, and inhibited cell M1 macrophage polarization. Our study indicates the anti-inflammatory properties of the PAs on SAP mice by decreasing ROS levels, suppressing NLRP3 inflammasome, and M1 macrophage polarization.
活性氧 (ROS) 的作用对于急性胰腺炎 (AP) 的发病机制至关重要。原花青素 (PAs) 已被证实具有抗氧化活性。我们的研究旨在确定 PAs 是否通过减少 ROS、抑制 NLRP3 炎性体和抑制 M1 巨噬细胞极化来缓解 SAP。我们使用 SAP 小鼠研究了 PAs 对胰腺组织病理损伤的保护作用。在炎症 RAW 264.7 细胞或由脂多糖 (LPS) 诱导的小鼠骨髓来源巨噬细胞 (BMDM) 中研究了 PAs 对巨噬细胞的作用。免疫荧光染色和/或 Western blot 分析用于评估巨噬细胞和胰腺组织中的 NLRP3 炎性体。细胞计数试剂盒-8 (CCK-8) 用于评估 PAs 对细胞活力的影响，流式细胞术用于确定 PAs 对 RAW 264.7 细胞 ROS 水平的影响。然后，我们使用流式细胞术或实时定量聚合酶链反应 (RT-qPCR) 评估 M1 巨噬细胞极化。PAs 给药可减轻 SAP 小鼠的胰腺炎症。PAs 抑制了胰腺组织中的 NLRP3 炎性体和 M1 巨噬细胞极化。我们还发现 PAs 没有细胞毒性，但可以降低 RAW 264.7 细胞中的 ROS 水平，下调巨噬细胞中的 NLRP3 炎性体，并抑制细胞 M1 巨噬细胞极化。我们的研究表明 PAs 通过降低 ROS 水平、抑制 NLRP3 炎性体和 M1 巨噬细胞极化来发挥对 SAP 小鼠的抗炎作用。
