Department of Paediatrics, Zhujiang Hospital of Southern Medical University, Guangzhou, 510282 PR China.
Department of Paediatrics, Zhujiang Hospital of Southern Medical University, Guangzhou, 510282 PR China.
Int Immunopharmacol. 2023 Oct;123:110758. doi: 10.1016/j.intimp.2023.110758. Epub 2023 Aug 8.
Sepsis-associated encephalopathy (SAE) is characterised by long-term cognitive impairment and psychiatric illness in sepsis survivors, associated with increased morbidity and mortality. There is a lack of effective therapeutics for SAE. Molecular hydrogen (H2) plays multiple roles in septic diseases by regulating neuroinflammation, reducing oxidative stress parameters, regulating signalling pathways, improving mitochondrial dysfunction, and regulating astrocyte and microglia activation. Here we report the protective effect of hydrogen-rich saline in the juvenile SAE rat model and its possible underlying mechanisms. Rats were injected intraperitoneally with lipopolysaccharide at a dose of 5 mg/kg to induce sepsis; Hydrogen-rich saline (HRS) was administered 1 h after LPS induction at a dose of 5 ml/kg and nigericin at 1 mg/kg 1 h before LPS injection. H&E staining for neuronal damage, TUNEL assay for detection of apoptotic cells, immunofluorescence, ELISA protocol for inflammatory cytokines and 8-OHdG determination and western blot analysis to determine the effect of HRS in LPS-induced septic rats. Rats treated with HRS showed decreased TNF-α and IL-1β expression levels. HRS treatment enhanced the activities of antioxidant enzymes (SOD, CAT and GPX) and decreased MDA and MPO activities. The number of MMP-9 and NLRP3 positive immunoreactivity cells decreased in the HRS-treated group. Subsequently, GFAP, IBA-1 and CD86 immunoreactivity were reduced, and CD206 increased after HRS treatment. 8-OHdG expression was decreased in the HRS-treated rats. Western blot analysis showed decreased NLRP3, ASC, caspase-1, MMP-2/9, TLR4 and Bax protein levels after HRS treatment, while Bcl-2 expression increased after HRS treatment. These data demonstrated that HRS attenuated neuroinflammation, NLRP3 inflammasome activation, neuronal injury, and mitochondrial damage via NLRP3/Caspase-1/TLR4 signalling in the juvenile rat model, making it a potential therapeutic agent in the treatment of paediatric SAE.
脓毒症相关性脑病（SAE）的特征是脓毒症幸存者长期认知障碍和精神疾病，与发病率和死亡率增加有关。目前尚无有效的 SAE 治疗方法。分子氢（H2）通过调节神经炎症、降低氧化应激参数、调节信号通路、改善线粒体功能以及调节星形胶质细胞和小胶质细胞激活，在脓毒症疾病中发挥多种作用。在这里，我们报告了富氢盐水（HRS）在幼年脓毒症相关性脑病大鼠模型中的保护作用及其可能的潜在机制。大鼠腹腔注射 5mg/kg 脂多糖（LPS）诱导脓毒症；HRS 在 LPS 诱导后 1 小时以 5ml/kg 的剂量给药， Nigericin 在 LPS 注射前 1 小时以 1mg/kg 的剂量给药。用 H&E 染色进行神经元损伤，TUNEL 检测凋亡细胞，免疫荧光，ELISA 法检测炎症细胞因子和 8-OHdG 测定以及 Western blot 分析，以确定 HRS 对 LPS 诱导的脓毒症大鼠的影响。用 HRS 治疗的大鼠 TNF-α和 IL-1β表达水平降低。HRS 治疗增强了抗氧化酶（SOD、CAT 和 GPX）的活性，并降低了 MDA 和 MPO 的活性。HRS 治疗组 MMP-9 和 NLRP3 阳性免疫反应细胞的数量减少。随后，HRS 治疗后 GFAP、IBA-1 和 CD86 的免疫反应性降低，CD206 增加。HRS 治疗后大鼠 8-OHdG 表达减少。Western blot 分析显示，HRS 治疗后 NLRP3、ASC、caspase-1、MMP-2/9、TLR4 和 Bax 蛋白水平降低，而 Bcl-2 表达增加。这些数据表明，HRS 通过 NLRP3/Caspase-1/TLR4 信号通路减轻了幼年大鼠模型中的神经炎症、NLRP3 炎性小体激活、神经元损伤和线粒体损伤，使其成为治疗儿科 SAE 的潜在治疗剂。
