铁皮石斛多糖通过调节肠菌代谢物对老年小鼠肠道黏膜屏障损伤的改善作用  

Ameliorating effect of Dendrobium officinale polysaccharides on intestinal mucosal barrier damage in elderly mice by regulating intestinal microbial metabolites

作　　者：李宏 王慧[2] 王隶书[3] 王超楠 徐晓浩 孙立伟[2] LI Hong;WANG Hui;WANG Lishu;WANG Chaonan;XU Xiaohao;SUN Liwei(School of Pharmacy,Changchun University of Chinese Medicine,Changchun 130117,China;Experimental Center,Affiliated Hospital,Changchun University of Chinese Medicine,Changchun 130021,China;New Drug Research and Development Center,Jilin Academy of Chinese Medicine,Changchun 130012,China)

机构地区：[1]长春中医药大学药学院,吉林长春130117 [2]长春中医药大学附属医院实验中心,吉林长春130021 [3]吉林省中医药科学院新药研究开发中心,吉林长春130012

出　　处：《吉林大学学报(医学版)》2025年第1期76-84,共9页Journal of Jilin University:Medicine Edition

基　　金：吉林省教育厅科学技术研究项目(JJKH20230984KJ);吉林省科技厅科技发展计划项目(YDZJ202301ZYTS446)。

摘　　要：目的:探讨铁皮石斛多糖(DOP)对老年小鼠肠道黏膜屏障损伤的保护作用,并阐明其可能的作用机制。方法:选择10只雌性5月龄C57BL/6小鼠作为年轻组,将20只雌性C57BL/6小鼠(15月龄)随机分为老年组和老年鼠DOP(200 mg·kg^(-1)·d^(-1))处理组(DOP组),每组10只。DOP组小鼠采用DOP灌胃,检测各组小鼠体质量、摄食量和悬挂时间;HE染色观察各组小鼠肠道和脾脏组织病理形态表现;免疫组织化学染色检测各组小鼠肠道组织中闭锁小带蛋白1(ZO-1)和黏蛋白2(MUC2)的表达情况。制备肠菌代谢物培养基(IBMM)干预秀丽线虫(C.elegans),将C.elegans随机分为年轻IBMM组(Young-IBMM)、老年IBMM组(Aged-IBMM)和DOP-IBMM组,采用免疫荧光法分析各组C.elegans第1和12天的肠道脂褐素累积水平;亮蓝染色检测各组C.elegans第1和12天的肠道渗漏情况。制备IBMM干预Caco-2细胞,将Caco-2细胞分为Young-IBMM组、Aged-IBMM组和DOP-IBMM组,采用Western blotting法检测各组Caco-2细胞中ZO-1、闭合蛋白(Occludin)、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、磷酸化肌球蛋白轻链(p-MLC)和肌球蛋白轻链激酶(MLCK)蛋白表达水平。结果:与年轻组比较,老年组小鼠体质量增加(P<0.05),摄食量降低(P<0.05),悬挂时间缩短(P<0.05);与老年组比较,DOP组小鼠体质量明显减轻(P<0.01),摄食量增加(P<0.05),悬挂时间明显延长(P<0.01)。HE染色,与年轻组比较,老年组小鼠肠黏膜厚度变薄,杯状细胞减少,肠绒毛长短不一且排列无序,脾脏表面可见大量铁血黄素存在,红髓内细胞成分减少,白髓内动脉周围淋巴鞘和淋巴小结残存或几乎消失;与老年组比较,DOP组小鼠肠黏膜厚度增加,杯状细胞增多,肠绒毛长度一致且排列整齐,脾脏红髓整体功能改善,白髓成分增加。免疫组织化学染色,与年轻组比较,老年组小鼠肠道组织中ZO-1和MUC2蛋白表达水平明显降低(P<0.05或P<0.001);与老年组比较,DOP组小鼠肠道组�Objective:To investigate the protective effect of Dendrobium officinale polysaccharides(DOP)on intestinal mucosal barrier damage,and to elucidate the possible mechanism.Methods:Ten female C57BL/6 mice,aged 5 months,were selected as young group;twenty femal C57BL/6 mice,aged 15 months,were randomly divided into aged group and DOP treatment group(200 mg·kg^(-1),DOP group),with 10 mice in each group.The mice in DOP group were administrated with DOP by gavage.The body mass,food intakes and hanging time of the mice in various groups were detected.HE staining was used to observe the pathomorphology of intestinal and spleen tissues of the mice in various groups.Immunohistochemical staining was used to detect the expressions of intestinal atresin 1(ZO-1)and Mucin 2(MUC2)in intestinal tissue of the mice in various groups.The intestinal baterial metabolite medium(IBMM)were prepared to intervene the Caenorhabditis elegans(C.elegans),and the C.elegans were randomly divided into Young-IBMM group,Aged-IBMM group,and DOP-IBMM group.Immunofluorescence method was used to analyze the intestinal lipofuscin accumulation levels on the 1st day and the 12th day of the C.elegans in various groups.Brilliant blue staining was used to assess the intestinal leakage on the 1st day and the 12th day of C.elegans in various groups.The Caco-2 cells were randomly divided into Young-IBMM,Aged-IBMM and DOP-IBMM groups,and Western blotting method was used to detect the expression levels of ZO-1,Occludin,tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),phosphorylated myosin light chain(p-MLC),myosin light chain kinase(MLCK)proteins in the Caco-2 cells in various groups.Results:Compared with young group,the body mass of the mice in aged group was increased(P<0.05),the amount of food intake was decreased(P<0.05),and the hanging time was decreased(P<0.05);compared with aged group,the body mass of the mice in DOP group was significantly decreased(P<0.01),the amount of food intake was increased(P<0.05),and the hanging time was significantly extended(

关 键 词：铁皮石斛多糖 自然衰老小鼠 肠道屏障 肠菌代谢物 

分 类 号：R965[医药卫生—药理学]