背景:缺乏耐甲氧西林金黄色葡萄球菌(MRSA)感染的有效治疗方法,这通常会导致严重的急性肺损伤(ALI),对人类生命构成严重威胁。苦参苷(SOP),一种富含于传统中药苦参果实中的异黄酮苷,对特应性皮炎有抗炎作用,过敏性炎症,和脂多糖诱导的ALI。然而,其对MRSA诱导的ALI的作用和潜在机制尚不清楚.
目的:本研究的目的是评估SOP在MRSA诱导的ALI中的保护作用,并阐明其潜在的分子机制。
方法:用野生型小鼠进行体内实验,建立MRSA诱导的ALI小鼠模型,通过苏木精-伊红染色评价SOP对ALI的影响,流式细胞术,定量实时聚合酶链反应,和几个生化指标。过继转移实验以及BTB和CNC同源性1敲除(Bach1-/-)小鼠也用于本研究。体外研究采用鼠巨噬细胞RAW264.7细胞,原代骨髓源性巨噬细胞(BMDMs),和原代肺巨噬细胞探讨潜在的分子机制。
结果:SOP的给药通过改善肺组织学损伤改善MRSA诱导的ALI,减少中性粒细胞浸润,抑制氧化应激水平,降低炎性细胞因子的表达。在ALI小鼠肺巨噬细胞的分离实验和巨噬细胞过继转移实验中,SOP阻止巨噬细胞活化,从而减少促炎细胞因子的产生。体外实验表明,SOP降低脂磷壁酸(LTA)刺激的RAW264.7细胞中炎症介质的表达,BMDM,和原发性肺巨噬细胞。此外,SOP抑制蛋白激酶B(Akt)磷酸化和用MK2206处理-Akt消除SOP抑制LTA刺激的巨噬细胞炎症能力的特异性抑制剂。此外,用LTA或MRSA刺激上调Bach1表达;然而,Bach1的缺失消除了SOP对p-Akt激活以及炎症和ALI发展的抑制作用。
结论:这项研究提供了第一个证据,即SOP通过抑制Bach1/Akt途径抑制巨噬细胞活化,从而有效减轻MRSA诱导的ALI。这些发现强调了SOP作为治疗MRSA诱导的ALI的新型治疗剂的潜力。
BACKGROUND: The lack of effective treatments for methicillin-resistant Staphylococcus aureus (MRSA) infection, which often leads to severe acute lung injury (ALI), poses a grave threat to human life. Sophoricoside (SOP), an isoflavone glycoside abundant in the fruit of traditional Chinese herbal Sophora japonica l., showed anti-inflammatory effects against atopic dermatitis, allergic inflammation, and lipopolysaccharide-induced ALI. However, its effect and underlying mechanism on MRSA-induced ALI remain unclear.
OBJECTIVE: The aim of this study is to assess the protective effect of SOP in MRSA-induced ALI and elucidate its underlying molecular mechanisms.
METHODS: In vivo experiments were conducted using wild-type mice to establish MRSA-induced ALI mouse model, and the effects of SOP on ALI were evaluated by hematoxylin-eosin staining, flow cytometry, quantitative real-time polymerase chain reaction, and several biochemical indicators. Adoptive transfer experiments and BTB and CNC homology 1 knockout (Bach1-/-) mice were also utilized in this study. In vitro studies employed murine macrophages RAW264.7 cells, primary bone marrow-derived macrophages (BMDMs), and primary lung macrophages to explore the underlying molecular mechanisms.
RESULTS: The administration of SOP ameliorated MRSA-induced ALI by improving pulmonary histological damages, reducing neutrophil infiltration, suppressing oxidative stress levels, and decreasing the expression of inflammatory cytokines. In isolation experiments with ALI mouse lung macrophages and macrophage adoptive transfer experiments, SOP prevented macrophage activation, thereby reducing the production of proinflammatory cytokines. In vitro experiments demonstrated that SOP decreased the expression of inflammatory mediators in lipoteichoic acid (LTA)-stimulated RAW264.7 cells, BMDMs, and primary lung macrophages. Additionally, SOP inhibited protein kinase B (Akt) phosphorylation and treatment with MK2206-a specific inhibitor of Akt-eliminated SOP\'s ability to suppress LTA-stimulated macrophage inflammation. Furthermore, stimulation with LTA or MRSA up-regulated Bach1 expression; however, deletion of Bach1 abolished the inhibitory effect of SOP on p-Akt activation as well as inflammation and ALI development.
CONCLUSIONS: This study provides the first evidence that SOP effectively mitigates MRSA-induced ALI via suppressing macrophage activation through the inhibition of Bach1/Akt pathway. These findings highlight the potential of SOP as a novel therapeutic agent for treating MRSA-induced ALI.