Liu Xinqiang, Ding Hongguang, Chen Miner, Li Xusheng, Xiao Yan, Han Yongli, Zeng Hongke
Department of Intensive Care Medicine, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou, 510800, People's Republic of China.
Department of Emergency Medicine, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou, 510800, People's Republic of China.
J Inflamm Res. 2024 Oct 14;17:7295-7310. doi: 10.2147/JIR.S481171. eCollection 2024.
Shenfu injection (SF) has demonstrated its potential to enhance cellular immunity and induce clinical regression in patients suffering from sepsis or infectious shock. However, the therapeutic effect of SF on sepsis-induced cognitive dysfunction (SAE) and the mechanisms involved are still unclear. We aimed to investigate the mechanism of SF in mice with SAE.
Sepsis was constructed by caecal ligation and puncture. Mice were injected intraperitoneally with SF or NLRP3 inhibitor. The hippocampus injury of brain tissues was evaluated, and the levels of inflammatory cytokines (IL-1β, IL-18) and NLRP3 and Caspase 1 were measured. The active ingredients of SF were analyzed using network pharmacology, and molecular docking of the active ingredients of SF with NLRP3 and Caspase 1 was performed. BV-2 cells were treated with LPS or norcoclaurine. CCK-8 detected the cell viability, and the levels of inflammatory cytokines and NLRP3 and Caspase 1 were measured.
SF and NLRP3 inhibitor increased survival rate and the number of crossing the platform and decreased the escape latency time of sepsis mice. Moreover, SF and NLRP3 inhibitor improved neuronal damage and apoptosis in hippocampus of sepsis mice. In addition, SF and NLRP3 inhibitor reduced the levels of inflammatory cytokines, as well as inflammasomes in sepsis mice. There were 43 active ingredients in SF. Among them, 22 were Renshen and 21 were Fuzi. Renshen and Fuzi, the main active components of SF, form a complex regulatory network with NLRP3 and Caspase 1. (R)-norcoclaurine was most closely bound to NLRP3 with binding energy of -7.2 kJ·mol, ignavine was most closely bound to Caspase 1 with binding energy of -8.3 kJ·mol. Norcoclaurine increased the cell viability and decreased inflammation and pyroptosis.
SF regulated NLRP3/Caspase 1 through (R)-norcoclaurinee to prevent SAE.
参附注射液（SF）已显示出增强细胞免疫及促使脓毒症或感染性休克患者临床症状缓解的潜力。然而，SF对脓毒症诱导的认知功能障碍（SAE）的治疗效果及相关机制仍不清楚。我们旨在研究SF对SAE小鼠的作用机制。
通过盲肠结扎和穿刺构建脓毒症模型。小鼠腹腔注射SF或NLRP3抑制剂。评估脑组织的海马损伤情况，检测炎性细胞因子（IL-1β、IL-18）水平以及NLRP3和半胱天冬酶-1的水平。运用网络药理学分析SF的活性成分，并对SF的活性成分与NLRP3和半胱天冬酶-1进行分子对接。用脂多糖（LPS）或去甲乌药碱处理小胶质细胞系BV-2细胞。采用CCK-8法检测细胞活力，并检测炎性细胞因子水平以及NLRP3和半胱天冬酶-1的水平。
SF和NLRP3抑制剂提高了脓毒症小鼠的存活率和穿越平台的次数，并缩短了逃避潜伏期。此外，SF和NLRP3抑制剂改善了脓毒症小鼠海马区的神经元损伤和凋亡。另外，SF和NLRP3抑制剂降低了脓毒症小鼠的炎性细胞因子水平以及炎性小体水平。SF中有43种活性成分。其中，22种来自人参，21种来自附子。人参和附子作为SF的主要活性成分，与NLRP3和半胱天冬酶-1形成复杂的调控网络。（R）-去甲乌药碱与NLRP3结合最紧密，结合能为-7.2 kJ·mol，鹅掌楸碱与半胱天冬酶-1结合最紧密，结合能为-8.3 kJ·mol。去甲乌药碱提高了细胞活力，减轻了炎症和细胞焦亡。
SF通过（R）-去甲乌药碱调节NLRP3/半胱天冬酶-1以预防SAE。
