文献类型：期刊文献

中文题名：黑逍遥散调节肠道屏障改善APP/PS1小鼠学习记忆能力的作用机制

英文题名：Mechanism of Hei Xiaoyaosan in Improving Learning and Memory Abilities of APP/PS1 Mice by Regulating Intestinal Barrier

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：181

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;肠道屏障;短链脂肪酸;tau蛋白

外文关键词：Hei Xiaoyaosan;Alzheimer's disease;intestinal barrier;short-chain fatty acid;tau protein

摘要：目的:探究黑逍遥散调节肠道屏障改善阿尔茨海默病(AD)模型小鼠学习记忆能力的作用机制。方法:将60只4月龄雄性APP/PS1双转基因小鼠随机分为模型组,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,同月龄雄性C57BL/6J小鼠10只作为空白组,连续给药90 d。Morris水迷宫试验进行行为学检测,苏木素-伊红(HE)染色分析小鼠结肠形态学变化,透射电镜(TEM)观察结肠黏膜屏障超微结构变化,液相色谱-质谱联用(LC-MS/MS)检测小鼠粪便中短链脂肪酸(SCFAs)的含量,蛋白免疫印迹法(Western blot)检测海马组织tau、磷酸化(p)-tau(Thr231)、p-tau(Ser262)、p-tau(Ser396)、p-tau(Thr181)及结肠组织密封蛋白-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,模型组小鼠逃避潜伏期显著延长(P<0.01),目标象限滞留时间百分比显著缩短(P<0.01),p-tau(Thr231)、p-tau(Ser262)、p-tau(Ser396)、p-tau(Thr181)蛋白表达显著提高(P<0.01),肠道黏膜层腺体皱缩,腺体数量明显减少,杯状细胞数量减少,固有层重度水肿,炎性细胞浸润,血管扩张,上皮细胞水样变性;微绒毛排列稀疏、溶解及断裂较为严重,少量细胞内容物渗出肠腔,胞质内高程度空泡数量较多,紧密连接之间的膜及连接蛋白溶解,桥粒连接两侧张力丝减少,丙酸、乙酸、丁酸、异戊酸含量明显下降(P<0.05,P<0.01),Claudin-1、Occludin和ZO-1蛋白表达显著降低(P<0.01);与模型组比较,黑逍遥散高剂量组逃避潜伏期显著缩短(P<0.01),目标象限滞留时间百分比显著延长(P<0.01),p-tau(Thr231)、p-tau(Ser262)、p-tau(Ser396)、p-tau(Thr181)蛋白表达水平显著下降(P<0.01),肠道组织中的固有层水肿逐渐减轻,腺体数量形态恢复,杯状细胞数量逐渐增多,微绒毛数量尚可,细胞间紧密连接、桥粒连接破坏较轻,丙酸、乙酸、丁酸、异丁酸含量明显提高(P<0.05,P<0.01),戊酸、异戊酸含量有所增高,但差异无统计学意义;Claudin-1、Occludin和ZO-1蛋白表达显著提高(P<0.01)。结论:黑逍遥散可改善AD小鼠学习认知能力,其机制可能与通过降低肠道屏障通透性,提高SCFAs含量,从而降低tau蛋白的过度磷酸化相关。
Objective:To explore the mechanism of action of Hei Xiaoyaosan in regulating the intestinal barrier and improving the learning and memory abilities of Alzheimer's disease(AD)model mice.Methods:Sixty four-month-old male amyloid precursor protein/presenilin 1(APP/PS1)double transgenic mice were randomly divided into a model group,low,medium,and high dose groups of Hei Xiaoyaosan(5.53,11.05,and 22.10 g·kg^(-1)),a group treated with bifidobacterium tetravaccine tablets(0.585 g·kg^(-1)),and a group treated with donepezil hydrochloride(6.5 mg·kg^(-1)),with 10 mice in each group.Additionally,10 male C57BL/6J mice of the same age were used as a blank control group.The treatment was administered continuously for 90 days.The Morris water maze test was performed for behavioral detection.Hematoxylin-eosin(HE)staining was employed to analyze the morphological changes of the colon in mice.Transmission electron microscopy(TEM)was utilized to observe the ultrastructural changes of the colonic mucosal barrier.Liquid chromatography-tandem mass spectrometry(LC-MS/MS)was used to detect the content of short-chain fatty acids(SCFAs)in the feces of mice.Western blot was carried out to detect the expression of related proteins,including tau,phosphorylation(p)-tau(Thr231),p-tau(Ser262),p-tau(Ser396),and p-tau(Thr181)in the hippocampal tissue,as well as Claudin-1,Occludin,and zonula occludens-1(ZO-1)in the colonic tissue.Results:Compared with those of the blank group,in the model group,the escape latency of mice was significantly prolonged(P<0.01),and the percentage of residence time in the target quadrant was significantly shortened(P<0.01).The expression of p-tau(Thr231),p-tau(Ser262),p-tau(Ser396),and p-tau(Thr181)proteins was significantly increased(P<0.01).In the intestine,the glands in the mucosal layer shrank,and the number of glands decreased severely.The number of goblet cells decreased.The lamina propria was severely edematous,and inflammatory cells infiltrated.The blood vessels dilated,and epithelial cells showed hydropic degeneration.The microvilli were arranged sparsely,and there was severe dissolution and breakage.A small amount of cellular contents exuded into the intestinal lumen,and there were a large number of highly vacuolated structures in the cytoplasm.The membranes between the tight junctions and the junction proteins dissolved,and the tonofilaments on both sides of the desmosome junctions decreased.The content of propionic acid,acetic acid,butyric acid,and isovaleric acid was significantly decreased(P<0.05,P<0.01),and the expression of Claudin-1,Occludin,and ZO-1 proteins was significantly reduced(P<0.01).Compared with those of the model group,in the high-dose group of Hei Xiaoyaosan,the escape latency was significantly shortened(P<0.01),and the percentage of residence time in the target quadrant was significantly prolonged(P<0.01).The expression of p-tau(Thr231),p-tau(Ser262),p-tau(Ser396),and p-tau(Thr181)proteins decreased significantly(P<0.01).In the intestinal tissue,the edema of the lamina propria gradually alleviated,and the number and morphology of the glands were restored.The number of goblet cells gradually increased,and the number of microvilli was acceptable.The damage to the tight junctions and desmosome junctions between cells was relatively mild.The content of propionic acid,acetic acid,butyric acid,and isobutyric acid was significantly increased(P<0.05,P<0.01).The content of valeric acid and isovaleric acid was increased,but there was no statistical significance.The expression of Claudin-1,Occludin,and ZO-1 proteins was significantly increased(P<0.01).Conclusion:Hei Xiaoyaosan can improve the learning and cognitive abilities of AD mice,possibly through reducing intestinal barrier permeability and increasing the content of SCFAs,thereby reducing the excessive phosphorylation of tau proteins.