Molecular Medicine Research Center, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, PR China; School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, PR China.
Molecular Medicine Research Center, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, PR China.
Phytomedicine. 2022 Nov;106:154427. doi: 10.1016/j.phymed.2022.154427. Epub 2022 Sep 1.
Liver dysfunction and liver failure are serious complications of sepsis, directly leading to septic progression and death. Now, there is no specific therapeutics available for sepsis-related liver dysfunction. Prim-O-glucosylcimifugin (POG), a chromone richest in the roots of Saposhnikovia divaricata (Turcz.) Schischk, is usually used to treat headache, rheumatoid arthritis and tetanus. While, the underlying mechanisms of POG against sepsis-induced liver damage and dysfunction are still not clear.
To study the anti-sepsis effect of POG, and its pharmacological mechanism to protect liver injury by weakening the function of macrophages in septic livers through inhibiting NOD-like receptor protein 3 (NLRP3) inflammasome pathway.
In vivo experiments, septic mouse model was induced by cecal ligation and puncture (CLP), and then the mortality was detected, liver inflammatory damages and plasma biomarkers of liver injury were evaluated by histopathological staining and biochemical assays, respectively. In vitro experiments, mouse primary peritoneal macrophages were treated with lipopolysaccharide (LPS) and ATP, and then the activated-inflammasomes, macrophage migration and polarization were detected by ASC immunofluorescence staining, transwell and flow cytometry assays, respectively. NLRP3 inflammasome components NLRP3, caspase-1, IL-1β and IL-18 protein expressions were detected using western blot assays, and the contents of IL-1β and IL-18 were measured by ELISA assays.
POG treatment significantly decreased the mortality, liver inflammatory damages, hepatocyte apoptosis and plasma biomarkers of liver injury in CLP-challenged male WT mice, which were comparable to those in ibuprofen (a putative anti-inflammatory drug)-supplemented septic male WT mice and septic NLRP3 deficient-male mice. POG supplementation significantly suppressed NLRP3 inflammasome activation in septic liver tissues and cultured macrophages, by significantly reducing NLRP3, cleaved-caspase-1, IL-1β and IL-18 levels, the activated-inflammasome ASC specks, and macrophage infiltration and migration, as well as M1-like polarization, but significantly increasing M2-like polarization. These findings were similar to the pharmacological effects of ibuprofen, NLRP3 deficiency, and a special NLRP3 inhibitor, MCC950.
POG protected against sepsis by inhibiting NLRP3 inflammasome-mediated macrophage activation in septic liver and attenuating liver inflammatory injury, indicating that it may be a potential anti-sepsis drug candidate.
肝功能障碍和肝功能衰竭是脓毒症的严重并发症，直接导致脓毒症的进展和死亡。目前，尚无针对脓毒症相关肝功能障碍的特效疗法。原-0-葡萄糖基升麻素（POG）是独活根部最丰富的色酮，通常用于治疗头痛、类风湿关节炎和破伤风。然而，POG 对脓毒症引起的肝损伤和功能障碍的潜在机制尚不清楚。
通过抑制脓毒症肝脏中巨噬细胞 NLRP3 炎性小体途径的功能，研究 POG 的抗脓毒症作用及其保护肝损伤的药理机制。
体内实验，采用盲肠结扎穿孔（CLP）法诱导脓毒症小鼠模型，检测死亡率，通过组织病理学染色和生化检测分别评估肝炎症损伤和血浆肝损伤生物标志物。体外实验，用脂多糖（LPS）和 ATP 处理小鼠原代腹腔巨噬细胞，通过 ASC 免疫荧光染色、transwell 和流式细胞术分别检测激活的炎性小体、巨噬细胞迁移和极化。用 Western blot 检测 NLRP3 炎性小体相关蛋白 NLRP3、caspase-1、IL-1β和 IL-18 的表达，用 ELISA 检测 IL-1β和 IL-18 的含量。
POG 治疗可显著降低 CLP 攻击的雄性 WT 小鼠的死亡率、肝炎症损伤、肝细胞凋亡和血浆肝损伤生物标志物，其效果与布洛芬（一种假定的抗炎药）补充的脓毒症雄性 WT 小鼠和脓毒症 NLRP3 缺陷型雄性小鼠相当。POG 补充可显著抑制脓毒症肝脏组织和培养的巨噬细胞中 NLRP3 炎性小体的激活，显著降低 NLRP3、cleaved-caspase-1、IL-1β和 IL-18 水平、激活的炎性小体 ASC 斑点、巨噬细胞浸润和迁移以及 M1 样极化，但显著增加 M2 样极化。这些发现与布洛芬、NLRP3 缺陷和一种特殊的 NLRP3 抑制剂 MCC950 的药理作用相似。
POG 通过抑制脓毒症肝脏中 NLRP3 炎性小体介导的巨噬细胞激活并减轻肝炎症损伤，从而防止脓毒症，表明其可能是一种有潜力的抗脓毒症药物候选物。
