Data Sheet 1_Multi-omics reveal the neuroprotective mechanisms of Xinshubao tablet against scopolamine-induced cognitive dysfunction in mice.zip

IntroductionAlzheimer’s disease (AD) is a progressive neurodegenerative disorder with limited treatments. Xinshubao tablet (XSB), a traditional Chinese medicine, contains several bioactive compounds with notable neuroprotective effects. Our previous studies have demonstrated that XSB can alleviate cognitive deficits in vascular dementia (VaD) models, suggesting its potential as a therapeutic candidate for AD.
MethodsIn this study, scopolamine-induced AD-like mice were orally administered with varying doses of XSB (0.13 g/kg, 0.26 g/kg and 0.52 g/kg) for 28 days. Behavior tests, H&E, Nissl, immunofluorescence staining, and Western blot assays were performed to evaluate the neuroprotection of XSB on AD-like mice. Then, fecal 16S rDNA sequencing, serum metabolomics, and hippocampal mRNA sequencing (mRNA-seq) analysis were performed to investigate the underlying mechanisms.
Results and discussionThe results revealed that oral administration of XSB improved cognitive function, mitigated neuropathological damage, and alleviated dysfunction in the cholinergic system in AD-like mice. XSB treatment also enhanced gut microbiota diversity, increased the abundance of Enterococcus, Actinobacteriota, Coriobacteriales, and Eggerthellaceae, but reduced the abundance of Helicobacter rodentium and Lachnospiraceae. Integrating mRNA-seq and metabolomics data highlighted key regulatory pathways including the biosynthesis of unsaturated fatty acids, tyrosine metabolism, and glycerophospholipid metabolism. Furthermore, XSB treatment reduced the expression of TNF-α, IL-1β, MPO, enhanced SOD, GSH activities, reduced malondialdehyde (MDA) levels, upregulated the expression of BDNF, SYN, PSD95, and improved synaptic density. Transformation of XSB derived fecal microbiota (XSB-FM) effectively alleviated cognitive dysfunction and intestinal barrier injures. In conclusion, XSB may exert its neuroprotective effects via the microbiota-metabolite-brain axis, thereby improving neuroinflammation, neurotransmission, and synaptic integrity. These findings support the potential of XSB as a multifactorial therapeutic strategy for cognitive deficits in AD.

阿尔茨海默病（Alzheimer’s disease, AD）是一种进展性神经退行性疾病，目前临床治疗手段有限。舒心宝片（Xinshubao tablet, XSB）作为一味中药，含有多种具备显著神经保护活性的生物活性成分。既往研究已证实，XSB可改善血管性痴呆（vascular dementia, VaD）模型小鼠的认知功能缺损，提示其有望成为AD治疗的潜在候选药物。
方法：本研究以东莨菪碱诱导的类AD小鼠为模型，连续28天灌胃给予不同剂量的XSB（0.13 g/kg、0.26 g/kg及0.52 g/kg）。通过行为学实验、苏木精-伊红（H&E）染色、尼氏染色、免疫荧光染色及蛋白质印迹实验，评估XSB对类AD小鼠的神经保护作用。此外，采用粪便16S rDNA测序、血清代谢组学及海马mRNA测序（mRNA-seq）分析，探究其潜在作用机制。
结果与讨论：结果显示，灌胃给予XSB可改善类AD小鼠的认知功能，减轻神经病理损伤，缓解胆碱能系统功能异常。XSB干预可提升肠道菌群多样性，增加肠球菌属（Enterococcus）、放线菌门（Actinobacteriota）、柯林斯菌目（Coriobacteriales）及埃格特菌科（Eggerthellaceae）的菌群丰度，同时降低啮齿类幽门螺杆菌（Helicobacter rodentium）与毛螺菌科（Lachnospiraceae）的丰度。整合mRNA-seq与代谢组学数据后，筛选得到关键调控通路，包括不饱和脂肪酸生物合成、酪氨酸代谢及甘油磷脂代谢。此外，XSB干预可降低肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）及髓过氧化物酶（MPO）的表达，提高超氧化物歧化酶（SOD）与谷胱甘肽（GSH）的活性，降低丙二醛（MDA）水平；同时上调脑源性神经营养因子（BDNF）、突触素（SYN）及突触后致密蛋白95（PSD95）的表达，改善突触密度。舒心宝片来源的粪便菌群移植（XSB-FM）可有效缓解认知功能障碍与肠屏障损伤。综上，XSB可能通过菌群-代谢物-脑轴发挥神经保护作用，进而改善神经炎症、神经传递与突触完整性。本研究结果支持XSB作为AD认知缺损的多靶点治疗策略的潜在应用价值。