【摘要】 目的:观察限时进食法干预高盐血瘀证大鼠的疗效，分析其对肠道代谢产物的影响，深入探讨限时进食法改善高盐血瘀证的具体作用机制。探讨限时进食法干预高盐血瘀证的机制。方法:将30只(雌雄各半)Wistar大鼠随机分成正常对照组、模型组和限时进食组，每组10只。正常对照组:自由进食普通饲料、饮水。模型组与限时进食组:进食含8%氯化钠的特制高盐饲料，自由饮水。限时进食组:在8 am-4 pm之间自由进食，其余时间不予进食，持续8周。处死大鼠，各组大鼠进行血液流变学分析、凝血四项检测、苏木精-伊红染色观察胸主动脉血管壁病变程度和结肠病理形态、采用16Sr DNA基因高通量测序检测肠道微生物的组成、利用UHPLC-QE-MS非靶向代谢组学方法，分析肠内容物的代谢物，以确定限时进食对肠道菌群代谢的影响。结果:和对照组相比，模型组全血黏度明显升高(P<0.001);血浆黏度明显升高(P<0.05);活化部分凝血活酶时间和凝血酶原时间显著缩短(P<0.05)，凝血酶时间和纤维蛋白原含量均显著延长(P<0.05);胸主动脉HE染色可见血管结构破坏，内皮脱落坏死，轻度炎细胞浸润;与模型组比较，限时进食组全血黏度显著降低(P<0.005);血浆黏度显著降低(P<0.01);活化部分凝血活酶时间和凝血酶原时间显著延长(P<0.05)，凝血酶时间和纤维蛋白原含量均显著缩短(P<0.05);与对照组比较，模型组厚壁菌门相对丰度显著降低(P<0.001)，拟杆菌菌门相对丰度显著升高(P<0.001)。与模型组比较，限时进食组厚壁菌门相对丰度显著升高(P<0.05)，拟杆菌菌门相对丰度显著降低(P<0.05)。与对照组相比较，模型组未培养拟杆菌目细菌、普雷沃菌科UCG＿001菌属/种、未分类丹毒丝菌科细菌、鼠杆菌属、苏黎世杆菌属、相对丰度显著增加(P<0.01)、未培养拟杆菌目细菌的相对丰度显著降低(P<0.01);限时进食法干预后高盐血瘀证大鼠粪便中光洁素A、1-羟基-5,5-二甲基吡咯烷-2-醇、4-乙烯基苯酚硫酸酯、落葵皂苷A、脱氧胸苷酸、甜菜碱、5-羟基吲哚乙酸、花生四烯酸表达显著上调(P<0.01); PA(8:0/18:1(12Z)-2OH9,10)表达显著下调(P<0.01)。结论:限时进食通过重塑肠道菌群结构(提升厚壁菌门/拟杆菌门比值)，促进保护性代谢物生成(如花生四烯酸抗炎、甜菜碱维持渗透压)，从而改善高盐诱导的血瘀状态，为限时进食法干预盐敏感性心血管疾病提供微生物-代谢物轴机制依据。更多还原

【Abstract】 Objective: To investigate the efficacy of time-restricted feeding( TRF) in ameliorating high-salt-induced blood stasis syndrome in rats,analyze its effects on intestinal metabolites,and explore the underlying mechanisms of TRF intervention. Methods:Thirty Wistar rats( 15 males and 15 females) were randomly divided into three groups( n = 10 per group) : normal control group,model group,and TRF group. The normal control group received a standard diet and water ad libitum. The model and TRF groups were fed a high-salt diet( containing 8% sodium chloride) with free access to water. The TRF group was allowed free access to food only between 8: 00 AM and 4: 00 PM for 8 weeks. After sacrifice,blood rheology,coagulation parameters( APTT,PT,TT,FIB),and pathological changes in the thoracic aorta and colon were evaluated using hematoxylin-eosin( H&E) staining. Gut microbiota composition was assessed via 16S r DNA high-throughput sequencing,and fecal metabolites were profiled using UHPLC-QE-MS-based untargeted metabolomics to determine the effects of TRF on gut microbiota metabolism. Results: Compared with the control group,the model group exhibited significantly increased whole blood viscosity( P < 0. 001) and plasma viscosity( P < 0. 05); significantly shortened APTT and PT( P < 0. 05); and significantly prolonged TT and increased FIB( P < 0. 05). H&E staining of the thoracic aorta revealed vascular structural damage,endothelial necrosis,and mild inflammatory cell infiltration in the model group. Compared to the model group,the TRF group showed significantly decreased whole blood viscosity( P < 0. 005) and plasma viscosity( P < 0. 01);significantly prolonged APTT and PT( P < 0. 05); and significantly shortened TT and decreased FIB( P < 0. 05). Microbiota analysis revealed a significant decrease in the relative abundance of Firmicutes( P < 0. 001) and a significant increase in Bacteroidetes( P <0. 001) in the model groupversus controls. TRF significantly increased the relative abundance of Firmicutes( P < 0. 05) and decreased Bacteroidetes( P < 0. 05) compared to the model group. Compared with the control group,the model groupshowed significantly increased relative abundances of " uncultured＿Bacteroidales＿bacterium",prevotellaceae＿UCG＿001,unclassified ＿Erysipelotrichaceae,Muribaculum,and Turicibacter( P < 0. 01),while the relative abundance of " uncultured＿Bacteroidales＿bacterium" was significantly decreased( P < 0. 01). TRF intervention significantly upregulated fecal metabolites in high-salt blood stasis model rats,including Glabrin A,1-hydroxy-5,5-dimethylpyrrolidin-2-ol,4-vinylphenol sulfate,Basellasaponin A,deoxythymidylic acid,betaine,5-hydroxyindoleacetic acid,and arachidonic acid( P < 0. 01),and significantly downregulated PA( 8: 0/18: 1( 12Z)-2OH9,10)( P < 0. 01).Conclusion: Time-restricted feeding ameliorates high-salt-induced blood stasis by remodeling gut microbiota structure( increasing the Firmicutes/Bacteroidetes ratio) and promoting the generation of protective metabolites( e. g. arachidonic acid for anti-inflammation,betaine for osmoregulation). This provides evidence for a gut microbiota-metabolite axis mechanism underlying TRF intervention in saltsensitive cardiovascular diseases.更多还原