岩藻多糖对酒精依赖小鼠肠黏膜屏障功能及神经行为学的作用  

Effects of fucoidan on intestinal mucosal barrier function and behaviors in alcohol-dependent mice

作　　者：张楠[1] 薛美兰[1] 高海琪 党凯 李捷锐 梁惠[2] ZHANG Nan;XUE Mei-lan;GAO Hai-qi;DANG Kai;LI Jie-rui;LIANG Hui(Basic Medical College,Qingdao University,Qingdao 266071,China;College of Public Health,Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学基础医学院,山东青岛266071 [2]青岛大学公共卫生学院,山东青岛266071

出　　处：《中国病理生理杂志》2022年第10期1790-1802,共13页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81872605);山东省自然科学基金资助项目(No.ZR2020MH215);山东省重大科技创新专项(No.2019JZZY010818)。

摘　　要：目的:探讨岩藻多糖对酒精依赖小鼠的肠黏膜屏障功能以及神经行为学的作用。方法:36只雄性C57BL/6J小鼠随机分为3组:对照组、模型组及岩藻多糖(300 mg/kg)干预组。模型组和岩藻多糖组动物于每周一至周五的上午8时至下午4时自由饮用新鲜自来水,然后自由饮用15%乙醇溶液至第2天上午8时。对照组的小鼠从周一到周五自由饮用自来水。所有小鼠周六切断一切水源,周日全天给予自来水自由饮用。实验持续10周。采用蔗糖偏好实验(sucrose preference test,SPT)、悬尾实验(tail suspension test,TST)、强迫游泳实验(forced swimming test,FST)、高架十字迷宫(elevated plus maze,EPM)实验和Y迷宫自由交替实验测定小鼠的行为和认知功能。采用ELISA法检测大脑皮质5-羟色胺(5-hydroxytryptamine,5-HT)、多巴胺(dopamine,DA)、神经肽Y(neuropeptide Y,NPY)、脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)和相关炎症因子水平。ELISA法检测血清内毒素、二胺氧化酶(diamine oxidase,DAO)、D-乳酸(D-lactic acid,D-LA)和脂肪酸结合蛋白2(fatty acid-binding protein 2,FABP2)水平。Western blot法和免疫荧光法测定紧密连接蛋白在回肠中的表达。用透射电镜观察小鼠回肠组织超微结构。用生物素示踪实验观察回肠组织通透性。结果:岩藻多糖能显著缓解酒精暴露小鼠的认知功能障碍和抑郁样行为。与对照组相比,模型组小鼠在SPT中显示对蔗糖溶液的偏好降低,Y迷宫实验有效进臂率降低,TST和FST的静止不动时间增加,EPM实验中开放臂进入时间减少(P<0.05)。与模型组比较,岩藻多糖治疗后小鼠对蔗糖溶液偏好度和开放臂进入时间增加,TST的固定时间减少(P<0.05)。岩藻多糖组小鼠脑BDNF和5-HT水平均高于模型组(P<0.05)。岩藻多糖可降低脑白细胞介素1β、肿瘤坏死因子α和单核细胞趋化因子1水平(P<0.05)。免疫荧光结果显示,岩藻多糖组小胶质细胞标AIM:To investigate the effects of fucoidan on intestinal mucosal barrier function and behaviors in alcohol-exposed mice.METHODS:Thirty-six male C57BL/6J mice were randomly divided into 3 groups:control group,model group and fucoidan(300 mg/kg)treatment group,with 12 mice in each group.The animals in the model group and fucoidan group were taken water freely from 8:00 AM to 4:00 PM,and switch to 15%alcohol solution from 4:00 PM to 8:00 AM for 10 weeks.The control mice were taken water freely.The behavioral and cognitive functions of mice were measured by sucrose preference test(SPT),tail suspension test(TST),forced swimming test(FST),elevated plus maze(EPM)test and Y-maze free alternate experiment.The levels of 5-hydroxytryptamine(5-HT),dopamine(DA),neuropeptide Y(NPY),brain-derived neurotrophic factor(BDNF)and inflammatory factors were detected by ELISA in cerebral cortex.The levels of serum endotoxin,diamine oxidase(DAO),D-lactic acid(D-LA)and fatty acid-binding protein2(FABP2)were measured by ELISA.The expression of tight junction proteins in the ileum was measured by Western blot and immunofluorescence.The ultrastructure of ileum tissues in mice was examined under transmission electron microscope.The permeability of ileum tissues was observed using tracer test.RESULTS:Fucoidan attenuated cognitive dysfunction and depression-like behaviors induced by alcohol in mice.Fucoidan increased the preference for sucrose solution in SPT and the time in open arm in EPM test,and reduced immobilization time in TST(P<0.05).Brain BDNF and 5-HT levels in fucoidan group were higher than those in model group(P<0.05).Fucoidan decreased the levels of brain interleukin-1β,tumor necrosis factor-αand monocyte chemokine protein-1(P<0.05).Immunofluorescence data showed that the expression levels of microglia marker CD68 and astrocyte marker glial fibrillary acidic protein(GFAP)were decreased in fucoidan group.Moreover,serum endotoxin,FABP2 and D-LA levels were decreased,the cell junction and permeability were improved,and the express

关 键 词：岩藻多糖 酒精中毒 抑郁 肠黏膜屏障 神经保护作用 

分 类 号：R363.1[医药卫生—病理学] R749[医药卫生—基础医学]