文献类型：期刊文献

中文题名：黑逍遥散对APP/PS1小鼠认知障碍及其肠道菌群的调节作用

英文题名：Regulatory Effect of Hei Xiaoyaosan on Cognitive Impairment and Gut Microbiota in APP/PS1 Mice

作者：陈怡琴[1];杨娇[1];裴文丽[1];韩玉梅[1];王虎平[1,2,3]

第一作者：陈怡琴

机构：[1]甘肃中医药大学,兰州730000;[2]甘肃省中医方药挖掘与创新转化重点实验室,兰州730000;[3]甘肃省中药新产品创制工程实验室,兰州730000

第一机构：甘肃中医药大学

年份：2025

卷号：31

期号：19

起止页码：191

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：;北大核心:【北大核心2023】；

基金：国家自然科学基金项目(82160862);第五批全国中医临床优秀人才研修项目(国中医药人教函[2022]239号);首批陇原青年英才项目(中共甘肃省委人才工作领导小组[2022]5号);甘肃省高校教师创新基金项目(2024A-083)。

语种：中文

中文关键词：黑逍遥散;阿尔茨海默病;肠道菌群;血脑屏障

外文关键词：Hei Xiaoyaosan;Alzheimer's disease;gut microbiota;blood-brain barrier

摘要：目的:观察黑逍遥散对阿尔茨海默病(AD)模型小鼠学习记忆能力的改善作用,探究黑逍遥散通过调节肠道菌群发挥抗AD的作用机制。方法:选取4月龄雄性C57BL/6J小鼠10只作空白组,同月龄雄性APP/PS1双转基因小鼠60只,随机分为模型组,黑逍遥散低、中、高剂量组(5.53、11.05、22.10 g·kg^(-1)),双歧杆菌四联活菌片组(0.585 g·kg^(-1))及盐酸多奈哌齐组(6.5 mg·kg^(-1)),每组10只,对应药物连续干预90 d。新物体识别实验观察各组小鼠学习记忆能力,苏木素-伊红(HE)染色分析小鼠海马CAI区形态学病理变化,16S rRNA测序技术检测小鼠粪便中的肠道菌群变化,透射电镜(TEM)观察血脑屏障超微结构变化,蛋白免疫印迹法(Western blot)检测密封蛋白-1(Claudin-1)、闭合蛋白(Occludin)、闭锁小带蛋白-1(ZO-1)相关蛋白的表达。结果:与空白组比较,模型组小鼠的新物体识别指数显著降低(P<0.01);神经元细胞数量减少,排列紊乱,胞核固缩,细胞结构不同程度的受损;血管内皮细胞膜广泛溶解,紧密连接间隙增大、膜溶解,连接蛋白减少,血管基膜少部分溶解、不连续,星胶足板基质水肿,线粒体水肿;Claudin-1、Occludin、ZO-1蛋白表达显著降低(P<0.01)。与模型组比较,黑逍遥散干预后,新物体识别指数明显提高(P<0.05,P<0.01);神经元细胞数量增多,排列较整齐,胞核固缩减少,胞体清晰,结构也比较完整;血管内皮细胞膜部分溶解,紧密连接间隙轻微增大,连接蛋白较多,血管基膜溶解减轻,星胶足板、线粒体水肿减轻;Claudin-1、Occludin和ZO-1蛋白表达明显提高(P<0.05,P<0.01)。肠道菌群分析结果显示,黑逍遥散可调节APP/PS1小鼠肠道菌群的结构和组成,与空白组比较,模型组厚壁菌门(Firmicutes)、乳杆菌属(Lactobacillus)、双歧杆菌属(Bifidobacterium)丰度明显降低(P<0.05,P<0.01),拟杆菌门(Bacteroidetes)、拟杆菌目S24-7属(unidentified_S24-7)丰度显著提高(P<0.01)。与模型组比较,黑逍遥散剂量组厚壁菌门(Firmicutes)、放线菌门(Actinobacteriota)、乳杆菌属(Lactobacillus)和双歧杆菌属(Bifidobacterium)丰度明显提高(P<0.05,P<0.01),拟杆菌门(Bacteroidetes)、拟杆菌目S24-7属(unidentified_S24-7)丰度明显降低(P<0.05,P<0.01)。结论:黑逍遥散可改善AD小鼠认知功能障碍和病理损伤,其机制可能与调整肠道菌群的丰度和结构,降低血脑屏障通透性,从而改善海马神经元损伤相关。
Objective:To observe the effect of Hei Xiaoyaosan on improving the learning and memory abilities of Alzheimer's disease(AD)model mice and explore the anti-AD mechanism of Hei Xiaoyaosan by regulating gut microbiota.Methods:Ten four-month-old male C57BL/6J mice were selected as a blank group,and 60 male amyloid precursor protein/presenilin 1(APP/PS1)double transgenic mice of the same age were randomly divided into a model group,low-dose,mediumdose,and high-dose groups of Hei Xiaoyaosan(5.53,11.05,22.10 g·kg^(-1)),a group of Bifidobacterium bifidum tetrapartum tablets(0.585 g·kg^(-1)),and a group of donepezil hydrochloride(6.5 mg·kg^(-1)),with 10 mice in each group.The continuous drug intervention was performed for 90 d.New object recognition experiments were performed to observe the learning and memory abilities of each group.Morphological pathological changes in the Cornu Ammonis 1(CA1)area of the hippocampus were analysed by hematoxylin-eosin(HE)staining,and the changes in gut microbiota in the feces of the mice were detected by 16S rRNA sequencing.Ultrastructural changes in the blood-brain barrier were observed by electron transmission endomicroscopy(TEM),and Western blot was employed to detect the expression of related proteins such as Claudin-1,Occludin,and zonula occludens-1(ZO-1).Results:Compared with that in the blank group,the new object recognition index was significantly decreased in the model group(P<0.01).The number of neuronal cells was reduced,and the arrangement was disordered.The cell nuclei were pyknosed.The cell structures were damaged to varying degrees.The vascular endothelial cell membranes were extensively dissolved.The tight junction gaps were enlarged,with the membranes dissolved,and the junction proteins were reduced.A small part of the vascular basement membrane was dissolved and discontinuous.The astrocytic endfeet matrix and mitochondria were edematous.The expression of Claudin-1,Occludin,and ZO-1 was significantly decreased(P<0.01).Compared with that in the model group,after the intervention with Hei Xiaoyaosan,the new object recognition index was significantly increased(P<0.05,P<0.01).The number of neuronal cells increased.The arrangement was neater.The pyknosis of cell nuclei was reduced.The cell bodies were clear,and the structures were relatively complete.The endothelial cell membranes were partially dissolved.The tight junction gaps were slightly enlarged.The junction proteins were relatively abundant.The dissolution of the vascular basement membranes was alleviated,and the swelling of the astrocytic endfeet and mitochondria was reduced.The expression of Claudin-1,Occludin,and ZO-1 was significantly increased(P<0.05,P<0.01).The results of the gut microbiota analysis showed that Hei Xiaoyaosan could regulate the structure and composition of the gut microbiota in APP/PS1 mice.Compared with the blank control group,the model group showed a significant decrease in the abundance of Firmicutes,Lactobacillus,and Bifidobacterium(P<0.05,P<0.01)and a significant increase in the abundance of Bacteroidetes and unidentified_S24-7(P<0.01).Compared with the model group,the Hei Xiaoyaosan groups showed a significant increase in the abundance of Firmicutes,Actinobacteriota,Lactobacillus,and Bifidobacterium(P<0.05,P<0.01)and a significant decrease in the abundance of Bacteroidetes and unidentified_S24-7(P<0.05,P<0.01).Conclusion:Hei Xiaoyaosan can improve cognitive dysfunction and pathological damage in AD mice.Its mechanism may be related to adjusting the abundance and structure of the gut microbiota,reducing the permeability of the blood-brain barrier,and thus improving hippocampal neuronal damage.