腹泻型肠易激综合征大鼠肠道菌群和肠道屏障功能的变化及相关分子机制的研究
发布时间：2021-06-30 21:25
目的:腹泻型肠易激综合征（diarrhea irritable bowel syndrome,IBS-D）是一种临床常见的功能性肠病,这种疾病的高发病率严重影响患者的生活质量,对卫生医疗资源造成很大的压力。IBS-D的病理生理学机制较为复杂。目前,其发病机制尚不明确。IBS-D患者常存在明显的胃肠动力异常,表现出轻度或者严重的腹泻并伴随出现腹痛等症状。研究发现肠道屏障功能的破坏以及肠道微生态的改变是其发病的重要原因。本实验旨在探讨IBS-D大鼠肠道微生态的组成改变以及与肠道屏障相关的紧密连接蛋白的变化,为IBS-D发病机制提供实验数据和理论基础。实验方法:将Wistar雄性大鼠随机分为对照组和模型组,对大鼠进行胃肠电极埋置手术,分别于胃窦、十二指肠、结肠部位埋置电极,检测两组大鼠胃肠肌电活动的改变。除此之外,对胃肠动力检测还应用了胃排空肠推进和结肠排珠实验;模型评价还包括腹壁撤退反射评分（abdominal withdrawal reflex,AWR）实验、HE染色、大鼠粪便含水量的检测;采用ELISA技术检测血浆中D-乳酸的含量;通过旷场实验以及糖水偏嗜实验评价IBS-D大鼠的焦虑抑...
【文章来源】：兰州大学甘肃省 211工程院校 985工程院校 教育部直属院校
【文章页数】：57 页
【学位级别】：硕士
【部分图文】：
腹泻型肠易激综合征大鼠胃肠动力的改变(a)IntestinalpropulsionandgastricemptyinginIBS-Drats.(b)ColonicbeadinginIBS-Drats.
兰州大学硕士学位论文腹泻型肠易激综合征大鼠肠道菌群和肠道屏障功能的变化及相关分子机制的研究24和细胞质染色，模型组结肠AQP3蛋白表达水平明显低于对照组。Figure6.ChangesofaquaporinsincolonofIBS-Drats.图6.腹泻型肠易激综合征大鼠结肠部位水通道蛋白的改变(a)ChangesofAQP3expressionincolon.(b)ChangesofAQP8expressionincolon.(c)ImmunohistochemistryforAQP3proteininthecolon(magnified100×scarbars:100μm;200×scarbars:50μm;and400×scarbars:50μm).Comparedtocontrol:*P<0.05,**P<0.01,***P<0.001.
兰州大学硕士学位论文腹泻型肠易激综合征大鼠肠道菌群和肠道屏障功能的变化及相关分子机制的研究253.7IBS-D紧密连接蛋白的改变qPCR和WesternBlot实验结果显示，与对照组相比IBS-D大鼠结肠组织中occludinmRNA以及蛋白质表达明显降低（P<0.01）,claudin-4mRNA以及蛋白质表达明显升高（P<0.05），模型组ZO-1mRNA的表达也显著降低（P<0.05）（Figure7.a,b,c,d,e）。免疫荧光染色显示claudin-4分布于肠上皮（包括细胞膜顶部）紧密结合的细胞膜中，模型组结肠中claudin-4荧光强度更高（Figure7.f）。Figure7.ChangesofTJproteinsinIBS-Drats.图7.腹泻型肠易激综合征大鼠紧密连接蛋白的改变(a)ThelevelofoccludinmRNAinhomogenatesofthecolon.(b)Thelevelofoccludinproteininhomogenatesofthecolon.(c)Thelevelofclaudin-4mRNAinthecolon.(d)Thelevelofclaudin-4proteininthecolon.(e)ThelevelofZO-1mRNAinhomogenatesofthecolon.(f)Immunofluorescencestainingdemonstratingthelocalizationofclaudin-4(red)(100×scarbars:100μm,200×scarbars:50μm,400×scarbars:50μm).Statisticalsignificancewasdeterminedcomparedtothecontrolgroup,*P<0.05,**P<0.01.
【参考文献】：
期刊论文
[1]New therapeutic perspectives in irritable bowel syndrome: Targeting low-grade inflammation, immuno-neuroendocrine axis, motility, secretion and beyond[J]. Emanuele Sinagra,Gaetano Cristian Morreale,Ghazaleh Mohammadian,Giorgio Fusco,Valentina Guarnotta,Giovanni Tomasello,Francesco Cappello,Francesca Rossi,Georgios Amvrosiadis,Dario Raimondo. World Journal of Gastroenterology. 2017(36)
[2]Visceral hypersensitivity in inflammatory bowel diseases and irritable bowel syndrome: The role of proteases[J]. Hannah Ceuleers,Hanne Van Spaendonk,Nikita Hanning,Jelena Heirbaut,Anne-Marie Lambeir,Jurgen Joossens,Koen Augustyns,Joris G De Man,Ingrid De Meester,Benedicte Y De Winter. World Journal of Gastroenterology. 2016(47)
[3]Gut microbiota role in irritable bowel syndrome: New therapeutic strategies[J]. Eleonora Distrutti,Lorenzo Monaldi,Patrizia Ricci,Stefano Fiorucci. World Journal of Gastroenterology. 2016(07)
[4]Irritable bowel syndrome:A clinical review[J]. Rosa LS Soares. World Journal of Gastroenterology. 2014(34)
[5]Microbiota-host interactions in irritable bowel syndrome: Epithelial barrier, immune regulation and brain-gut interactions[J]. Niall P Hyland,Eamonn MM Quigley,Elizabeth Brint. World Journal of Gastroenterology. 2014(27)
[6]Intestinal microbiota in pathophysiology and management of irritable bowel syndrome[J]. Kang Nyeong Lee,Oh Young Lee. World Journal of Gastroenterology. 2014(27)
[7]Multispecies probiotic protects gut barrier function in experimental models[J]. Mylene Nébot-Vivinus,Cherryl Harkat,Hanene Bzioueche,Christel Cartier,Raffaella Plichon-Dainese,Lara Moussa,Helene Eutamene,Dorsa Pishvaie,Sophie Holowacz,Christian Seyrig,Thierry Piche,Vassilia Theodorou. World Journal of Gastroenterology. 2014(22)
[8]Psychosocial determinants of irritable bowel syndrome[J]. Teodora Surdea-Blaga,Adriana Bban,Dan L Dumitrascu. World Journal of Gastroenterology. 2012(07)
[9]Expression of aquaporin 8 in colonic epithelium with diarrhoeapredominant irritable bowel syndrome[J]. WANG Jun-ping HOU Xiao-hua Department of Gastroenterology,Union Hospital of Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022,China. Chinese Medical Journal. 2007(04)
[10]腹泻型肠易激综合征患者临床特征与结肠黏膜水通道蛋白8的表达[J]. 王俊平,侯晓华,马瑞军. 中华内科杂志. 2006(12)
本文编号：3258525
【文章来源】：兰州大学甘肃省 211工程院校 985工程院校 教育部直属院校
【文章页数】：57 页
【学位级别】：硕士
【部分图文】：
腹泻型肠易激综合征大鼠胃肠动力的改变(a)IntestinalpropulsionandgastricemptyinginIBS-Drats.(b)ColonicbeadinginIBS-Drats.
兰州大学硕士学位论文腹泻型肠易激综合征大鼠肠道菌群和肠道屏障功能的变化及相关分子机制的研究24和细胞质染色，模型组结肠AQP3蛋白表达水平明显低于对照组。Figure6.ChangesofaquaporinsincolonofIBS-Drats.图6.腹泻型肠易激综合征大鼠结肠部位水通道蛋白的改变(a)ChangesofAQP3expressionincolon.(b)ChangesofAQP8expressionincolon.(c)ImmunohistochemistryforAQP3proteininthecolon(magnified100×scarbars:100μm;200×scarbars:50μm;and400×scarbars:50μm).Comparedtocontrol:*P<0.05,**P<0.01,***P<0.001.
兰州大学硕士学位论文腹泻型肠易激综合征大鼠肠道菌群和肠道屏障功能的变化及相关分子机制的研究253.7IBS-D紧密连接蛋白的改变qPCR和WesternBlot实验结果显示，与对照组相比IBS-D大鼠结肠组织中occludinmRNA以及蛋白质表达明显降低（P<0.01）,claudin-4mRNA以及蛋白质表达明显升高（P<0.05），模型组ZO-1mRNA的表达也显著降低（P<0.05）（Figure7.a,b,c,d,e）。免疫荧光染色显示claudin-4分布于肠上皮（包括细胞膜顶部）紧密结合的细胞膜中，模型组结肠中claudin-4荧光强度更高（Figure7.f）。Figure7.ChangesofTJproteinsinIBS-Drats.图7.腹泻型肠易激综合征大鼠紧密连接蛋白的改变(a)ThelevelofoccludinmRNAinhomogenatesofthecolon.(b)Thelevelofoccludinproteininhomogenatesofthecolon.(c)Thelevelofclaudin-4mRNAinthecolon.(d)Thelevelofclaudin-4proteininthecolon.(e)ThelevelofZO-1mRNAinhomogenatesofthecolon.(f)Immunofluorescencestainingdemonstratingthelocalizationofclaudin-4(red)(100×scarbars:100μm,200×scarbars:50μm,400×scarbars:50μm).Statisticalsignificancewasdeterminedcomparedtothecontrolgroup,*P<0.05,**P<0.01.
【参考文献】：
期刊论文
[1]New therapeutic perspectives in irritable bowel syndrome: Targeting low-grade inflammation, immuno-neuroendocrine axis, motility, secretion and beyond[J]. Emanuele Sinagra,Gaetano Cristian Morreale,Ghazaleh Mohammadian,Giorgio Fusco,Valentina Guarnotta,Giovanni Tomasello,Francesco Cappello,Francesca Rossi,Georgios Amvrosiadis,Dario Raimondo. World Journal of Gastroenterology. 2017(36)
[2]Visceral hypersensitivity in inflammatory bowel diseases and irritable bowel syndrome: The role of proteases[J]. Hannah Ceuleers,Hanne Van Spaendonk,Nikita Hanning,Jelena Heirbaut,Anne-Marie Lambeir,Jurgen Joossens,Koen Augustyns,Joris G De Man,Ingrid De Meester,Benedicte Y De Winter. World Journal of Gastroenterology. 2016(47)
[3]Gut microbiota role in irritable bowel syndrome: New therapeutic strategies[J]. Eleonora Distrutti,Lorenzo Monaldi,Patrizia Ricci,Stefano Fiorucci. World Journal of Gastroenterology. 2016(07)
[4]Irritable bowel syndrome:A clinical review[J]. Rosa LS Soares. World Journal of Gastroenterology. 2014(34)
[5]Microbiota-host interactions in irritable bowel syndrome: Epithelial barrier, immune regulation and brain-gut interactions[J]. Niall P Hyland,Eamonn MM Quigley,Elizabeth Brint. World Journal of Gastroenterology. 2014(27)
[6]Intestinal microbiota in pathophysiology and management of irritable bowel syndrome[J]. Kang Nyeong Lee,Oh Young Lee. World Journal of Gastroenterology. 2014(27)
[7]Multispecies probiotic protects gut barrier function in experimental models[J]. Mylene Nébot-Vivinus,Cherryl Harkat,Hanene Bzioueche,Christel Cartier,Raffaella Plichon-Dainese,Lara Moussa,Helene Eutamene,Dorsa Pishvaie,Sophie Holowacz,Christian Seyrig,Thierry Piche,Vassilia Theodorou. World Journal of Gastroenterology. 2014(22)
[8]Psychosocial determinants of irritable bowel syndrome[J]. Teodora Surdea-Blaga,Adriana Bban,Dan L Dumitrascu. World Journal of Gastroenterology. 2012(07)
[9]Expression of aquaporin 8 in colonic epithelium with diarrhoeapredominant irritable bowel syndrome[J]. WANG Jun-ping HOU Xiao-hua Department of Gastroenterology,Union Hospital of Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022,China. Chinese Medical Journal. 2007(04)
[10]腹泻型肠易激综合征患者临床特征与结肠黏膜水通道蛋白8的表达[J]. 王俊平,侯晓华,马瑞军. 中华内科杂志. 2006(12)
本文编号：3258525
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