目的 通过持续光照4周复制睡眠-觉醒昼夜节律障碍（circadian rhythm sleep-wake disorder，CRSWD）模型，探讨炒酸枣仁对CRSWD小鼠的作用及作用机制。方法 采用雄性C57BL/6J小鼠，持续光照4周复制CRSWD模型，给予炒酸枣仁进行干预，期间测定小鼠的体质量和摄食量，实验结束后分4个时间点（ZT0、ZT6、ZT12、ZT18）取材，利用qRT-PCR检测小鼠下丘脑时钟基因（circadian locomotor output cycles kaput，Clock）、脑和肌肉ARNT样蛋白1基因（brain-muscle arnt-like protein 1，Bmal1）、周期基因1（period 1，Per1）、Per2、隐花色素基因1（cryptochrom 1，Cry1）、视黄酸受体相关孤儿受体基因（retinoic acid receptor-related orphan，ROrα）、孤束核基因（reverse erythroblastosis virus，Rev-erbα）的表达；利用16S rRNA测序和GC-MS技术研究小鼠肠道菌群及短链脂肪酸含量的变化；采用UHPLC-Q-TOF-MS/MS技术研究模型小鼠大脑差异代谢物的变化。结果 炒酸枣仁显著调控模型小鼠下丘脑中Clock、Bmal1、Per1、Per2、Cry1、ROrα、Rev-erbα 7个生物钟基因的表达（P＜0.05、0.01、0.001），使其恢复到与空白组昼夜节律振荡趋势一致。肠道菌群16S rRNA测序与短链脂肪酸含量测定结果表明，炒酸枣仁可能通过升高梭状芽孢杆菌属Clostridium、乳酸杆菌属Lactobacillus、罗氏菌属Roseburia相对丰度，从而增加丁酸含量。脑代谢组学结果表明，ZT6和ZT18时共有的代谢通路有三羧酸循环、半胱氨酸和蛋氨酸代谢、烟酸和烟酰胺代谢。炒酸枣仁可以通过调节梭状芽孢杆菌属、乳酸杆菌属及其代谢产物丁酸的含量，进一步影响体内氨基酸代谢通路，从而恢复CRSWD小鼠模型睡眠-觉醒昼夜节律。结论 从节律生物钟基因水平、肠道菌群、脑代谢组学等多角度，揭示了炒酸枣仁治疗CRSWD小鼠模型的作用。

Objective To investigate the effect and mechanism of fried Suanzaoren (Ziziphi Spinosae Semen) on circadian rhythm sleep-wake disorder (CRSWD) mice by replicating CRSWD model with continuous light exposure for four weeks. Methods C57BL/6J male mice were exposed to continuous light for four weeks to replicate CRSWD model, fried Ziziphi Spinosae Semen was given for intervention, during which the body weight and food intake of mice were measured. After the experiment, samples were collected at four time points (ZT0, ZT6, ZT12, ZT18). Expressions of circadian locomotor output cycles kaput (Clock), brain-muscle arnt-like protein 1 (Bmal1), period 1 (Per1), Per2, cryptochrom 1 (Cry1), retinoic acid receptor-related orphan (ROrα) and reverse erythroblastosis virus (Rev-erbα) genes in hypothalamus of mice were detected by qRT-PCR, 16S rRNA sequencing and GC-MS were used to study the changes in gut microbiota and contents of short-chain fatty acids in mice, UHPLC-Q-TOF-MS/MS was used to study the changes of brain metabolites in model mice. Results Fried Ziziphi Spinosae Semen significantly regulated the expressions of seven circadian clock genes such as Clock, Bmal1, Per1, Per2, Cry1, ROrα and Rev-erbα in hypothalamus of model mice (P < 0.05, 0.01, 0.001), restored their circadian rhythm oscillation trend to be consistent with that of the blank group. Results of 16S rRNA sequencing of gut microbiota and determination of short chain fatty acid content indicated that fried Ziziphi Spinosae Semen may increase the content of butyric acid by increasing the relative abundance of Clostridium, Lactobacillus and Roseburia. The results of brain metabolomics indicated that the common metabolic pathways of ZT6 and ZT18 were tricarboxylic acid cycle, L-cysteine and methionine metabolism, niacin and nicotinamide metabolism. Fried Ziziphi Spinosae Semen could further affect the amino acid metabolism pathway in the body by regulating the abundance of Clostridium, Lactobacillus and their metabolite butyric acid content, thereby restoring the sleep wake circadian rhythm of CRSWD mice. Conclusion The therapeutic effect of fried Ziziphi Spinosae Semen on CRSWD mice model was revealed from multiple perspectives, such as the level of circadian clock genes, gut microbiota and brain metabolomics.

R285.5

山西省科学技术厅山西省科技创新人才团队项目（202304051001020）;山西省科学技术厅中央引导地方科技发展资金项目（YDZJSX2021CO25）;山西省科学技术厅自然科学研究面上项目（20210302123237）;山西省中医药管理局十大晋药资源评价与生态种植团队（zyytd2024026）;山西省中医药重点研究室建设项目（zyyyjs2024024）;山西省科技成果转化引导专项计划项目（202204021301063）;山西省基础研究计划自由探索类青年项目（20210302124279）