背景:肠脑轴(GBA)在脑缺血再灌注损伤(CIRI)中起着重要作用。大黄,以其泻药特性而闻名,已证明对CIRI具有保护作用。然而,目前尚不清楚这种保护作用是否通过GBA的调节来实现.
目的:本研究旨在探讨大黄提取物通过调节GBA途径改善CIRI的机制。
方法:我们使用LC-MS/MS鉴定了大黄提取物的活性成分。建立大脑中动脉闭塞(MCAO)模型,评价大黄提取物的作用。我们进行了16SrDNA测序和非靶向代谢组学来分析肠道内容物。此外,我们用HE染色,TUNEL染色,westernblot,和ELISA评估肠屏障完整性。我们通过ELISA测量血清中炎性细胞因子的水平。我们还使用伊文思蓝(EB)渗透检查了血脑屏障(BBB)的完整性,透射电子显微镜(TEM),westernblot,和ELISA。神经功能评分和TTC染色用于评估神经学结果。
结果:我们确定了大黄中的26种活性成分。大黄提取物增强了α-多样性,降低了肠杆菌科细菌的丰度,和部分纠正CIRI大鼠的代谢紊乱。它还改善了病理变化,增加了ZO-1、Occludin、和结肠中的Claudin1,并降低血清中LPS和d-lac的水平。此外,它降低了IL-1β的水平,血清中IL-6、IL-10、IL-17和TNF-α。大黄提取物减轻BBB功能障碍,EB渗透减少和海马微结构改善证明了这一点。它上调了ZO-1、Occludin、Claudin1,同时下调TLR4,MyD88和NF-κB的表达。同样,大黄提取物降低IL-1β水平,海马中的IL-6和TNF-α。最终,它降低了神经功能评分和脑梗死体积。
结论:大黄可有效治疗CIRI,可能通过抑制有害细菌,纠正代谢紊乱,修复肠屏障功能,缓解BBB功能障碍,并最终改善神经系统的结果。
BACKGROUND: The gut-brain axis (GBA) plays a central role in cerebral ischaemia-reperfusion injury (CIRI). Rhubarb, known for its purgative properties, has demonstrated protective effects against CIRI. However, it remains unclear whether this protective effect is achieved through the regulation of the GBA.
OBJECTIVE: This study aims to investigate the mechanism by which rhubarb extract improves CIRI by modulating the GBA pathway.
METHODS: We identified the active components of rhubarb extract using LC-MS/MS. The model of middle cerebral artery occlusion (MCAO) was established to evaluate the effect of rhubarb extract. We conducted 16S rDNA sequencing and untargeted metabolomics to analyze intestinal contents. Additionally, we employed HE staining, TUNEL staining, western blot, and ELISA to assess intestinal barrier integrity. We measured the levels of inflammatory cytokines in serum via ELISA. We also examined blood-brain barrier (BBB) integrity using Evans blue (EB) penetration, transmission electron microscopy (TEM), western blot, and ELISA. Neurological function scores and TTC staining were utilized to evaluate neurological outcomes.
RESULTS: We identified twenty-six active components in rhubarb. Rhubarb extract enhanced α-diversity, reduced the abundance of Enterobacteriaceae, and partially rectified metabolic disorders in CIRI rats. It also ameliorated pathological changes, increased the expressions of ZO-1, Occludin, and Claudin 1 in the colon, and reduced levels of LPS and d-lac in serum. Furthermore, it lowered the levels of IL-1β, IL-6, IL-10, IL-17, and TNF-α in serum. Rhubarb extract mitigated BBB dysfunction, as evidenced by reduced EB penetration and improved hippocampal microstructure. It upregulated the expressions of ZO-1, Occludin, Claudin 1, while downregulating the expressions of TLR4, MyD88, and NF-κB. Similarly, rhubarb extract decreased the levels of IL-1β, IL-6, and TNF-α in the hippocampus. Ultimately, it reduced neurological function scores and cerebral infarct volume.
CONCLUSIONS: Rhubarb effectively treats CIRI, potentially by inhibiting harmful bacteria, correcting metabolic disorders, repairing intestinal barrier function, alleviating BBB dysfunction, and ultimately improving neurological outcomes.