肠易激综合征模型大鼠结肠肌间神经丛钙视网膜蛋白阳性神经元增加
|
摘要
目的检测肠易激综合征腹泻型(IBS-D)和便秘型(IBS-C)两种模型大鼠远端回肠、结肠黏膜下神经丛(SMP)和肌间神经丛(MP)钙视网膜蛋白(CALR)阳性神经元的变化,探讨其在IBS发病中的作用。方法分别采用慢急性联合应激、冰水灌胃法建立IBS-D和IBS-C大鼠模型,制作回肠和结肠SMP、MP全层铺片标本,免疫组织荧光双染法检测CALR阳性神经元的改变。结果与对照组相比,IBS-D和IBS-C模型大鼠回肠SMP、MP和结肠SMP中CALR阳性神经元比例的差异无统计学意义(P>0.05),结肠MP中CALR阳性神经元的比例显著增加,分别为(17.2±3.2)%vs(13.4±2.9)%、(19.1±5.9)%vs(14.9±3.4)%,差异均有统计学意义(P<0.05)。结论 IBS-D、IBS-C模型大鼠结肠MP中CALR阳性神经元增加可能参与IBS肠道敏感性增高。
Objective To investigate the changes of calretinin-immunoreactive(CALR-IR) enteric neurons of submucosal plexus(SMP) and myenteric plexus(MP) in distal ileum and colon in rat models of irritable bowel syndrome with predominant diarrhea(IBS-D) and with predominant constipation(IBS-C). Methods Rat models of IBS-D were established with unpredictable mild stress and acute wrap restrain stress(CAS) and IBS-C were induced by administering cold water. Double immunohistochemical fluorescence staining with whole-mount preparations of SMP and MP was used to detect the changes in neurons with positive CALR-IR and their co-localization with the pan neuronal marker, anti-Hu. Results There was no statistically difference in the proportions of CALR-IR neurons in ileal SMP and MP in IBS-D and IBS-C model rats compared with corresponding control group(P>0.05). The similar results were found in colonic SMP in both IBS groups. While in colonic MP, the proportions of CALR-IR positive neurons in IBS-D model rats and IBS-C model rats were significantly increased compared with corresponding control group [(17.2±3.2)% vs(13.4±2.9)%,(19.1±5.9)% vs(14.9±3.4)%, P<0.05)]. Conclusion The increases of CALR-IR sensory neurons in MP of the IBS modeling rats may be involved in the visceral hypersensitivity of the IBS.
Objective To investigate the changes of calretinin-immunoreactive(CALR-IR) enteric neurons of submucosal plexus(SMP) and myenteric plexus(MP) in distal ileum and colon in rat models of irritable bowel syndrome with predominant diarrhea(IBS-D) and with predominant constipation(IBS-C). Methods Rat models of IBS-D were established with unpredictable mild stress and acute wrap restrain stress(CAS) and IBS-C were induced by administering cold water. Double immunohistochemical fluorescence staining with whole-mount preparations of SMP and MP was used to detect the changes in neurons with positive CALR-IR and their co-localization with the pan neuronal marker, anti-Hu. Results There was no statistically difference in the proportions of CALR-IR neurons in ileal SMP and MP in IBS-D and IBS-C model rats compared with corresponding control group(P>0.05). The similar results were found in colonic SMP in both IBS groups. While in colonic MP, the proportions of CALR-IR positive neurons in IBS-D model rats and IBS-C model rats were significantly increased compared with corresponding control group [(17.2±3.2)% vs(13.4±2.9)%,(19.1±5.9)% vs(14.9±3.4)%, P<0.05)]. Conclusion The increases of CALR-IR sensory neurons in MP of the IBS modeling rats may be involved in the visceral hypersensitivity of the IBS.
引文
[1]LI S,FEI G,FANG X,et al.Changes in enteric neurons of small intestine in a rat model of irritable bowel syndrome with diarrhea[J].J Neurogastroenterol Motil,2016,22(2):310-320.DOI:10.5056/jnm15082.
[2]陈玲玲,文平,费贵军,等.腹泻型肠易激综合征患者血清抗肠神经元抗体免疫反应特点及其临床意义[J].基础医学与临床,2017,37(2):156-161.DOI:10.3969/j.issn.1001-6325.2017.02.007.CHEN L L,WEN P,FEI G J,et al.Characteristics and clinical significance of sera anti-enteric neuronal antibodies in patients with irritable bowel syndrome with diarrhea[J].Basic&Clinical Medicine,2017,37(2):156-161.DOI:10.3969/j.issn.1001-6325.2017.02.007.
[3]李珊,费贵军,方秀才,等.腹泻型肠易激综合征模型大鼠结肠黏膜下神经丛神经元的改变[J].中华消化杂志,2010,30(4):250-253.DOI:10.3760/cma.j.issn.0254-1432.2010.04.008.LI S,FEI G J,FANG X C,et al.The morphological changes of colonic submucosal plexus in rats of irritable bowel syndrome with diarrhea[J].Chin J Dig,2010,30(4):250-253.DOI:10.3760/cma.j.issn.0254-1432.2010.04.008.
[4]MANN P T,SOUTHWELL B R,DING Y Q,et al.Localisation of neurokinin 3(NK3)receptor immunoreactivity in the rat gastrointestinal tract[J].Cell Tissue Res,1997,289(1):1-9.
[5]吕红,钱家鸣,金光亮,等.肠易激综合征慢急性联合应激动物模型的建立及其感觉、动力和心理行为的评价[J].中华内科杂志,2009,48(12):1035-1039.DOI:10.3760/cma.j.issn.0578-1426.2009.12.015.LYU H,QIAN J M,JIN G L,et al.The establishment of an animal model of gut-brain interaction in irritable bowel syndrome for the evaluation of visceral sensation,motility and psychological behavior[J].Chin J Intern Med,2009,48(12):1035-1039.DOI:10.3760/cma.j.issn.0578-1426.2009.12.015.
[6]彭丽华,杨云生,孙刚,等.便秘型肠易激综合征新概念模型的建立[J].世界华人消化杂志,2004,12(1):112-116.DOI:10.3969/j.issn.1009-3079.2004.01.029.PENG L H,YANG Y S,SUN G,et al.A new model of constipation-predominant irritable bowel syndrome in rats[J].World Chin J Digestol,2004,12(1):112-116.DOI:10.3969/j.issn.1009-3079.2004.01.029.
[7]杨希林,方秀才,刘晓红,等.便秘型肠易激综合征大鼠胃排空及胃内肌间神经丛神经元的变化[J].基础医学与临床,2011,31(12):1301-1304.YANG X L,FANG X C,LIU X H,et al.The changes of gastric emptying and gastric myenteric neurons in rats with IBS complicated with constipation[J].Basic&Clinical Medicine,2011,31(12):1301-1304.
[8]孙巧丽,袁建业,谢建群.肠易激综合征动物模型的研究现状和展望[J].中华消化杂志,2012,32(2):140-142.DOI:10.3760/cma.j.issn.0254-1432.2012.02.020.
[9]SCHWALLER B.Calretinin:from a“simple”Ca(2+)buffer to a multifunctional protein implicated in many biological processes[J].Front Neuroanat,2014,8:3.DOI:10.3389/fnana.2014.00003.
[10]ORD EZ N G.Value of calretinin immunostaining in diagnostic pathology:a review and update[J].Appl Immunohistochem Mol Morphol,2014,22(6):401-415.DOI:10.1097/PAI.0b013e31829b6fbd.
[11]GAO N,LUO J,URAY K,et al.CaMKⅡis essential for the function of the enteric nervous system[J].PLo S One,2012,7(8):e44426.DOI:10.1371/journal.pone.0044426.
[12]MITSUI R.Immunohistochemical characteristics of submucosal Dogiel typeⅡneurons in rat colon[J].Cell Tissue Res,2010,340(2):257-265.DOI:10.1007/s00441-010-0954-z.
[13]BEUSCHER N,JABARI S,STREHL J,et al.What neurons hide behind calretinin immunoreactivity in the human gut?[J].Histochem Cell Biol,2014,141(4):393-405.DOI:10.1007/s00418-013-1163-0.
[14]ZETZMANN K,STREHL J,GEPPERT C,et al.Calbindin D28kimmunoreactivity in human enteric neurons[J].Int J Mol Sci,2018,19(1):pii:E194.DOI:10.3390/ijms19010194.
[15]JAHNG J,KIM Y S.Irritable bowel syndrome:is it really a functional disorder?A new perspective on alteration of enteric nervous system[J].J Neurogastroenterol Motil,2016,22(2):163-165.DOI:10.5056/jnm16043.
[16]LTT A,MICHEL K,KRGER D,et al.High prevalence and functional effects of serum antineuronal antibodies in patients with gastrointestinal disorders[J].Neurogastroenterol Motil,2018,30(6):e13292.DOI:10.1111/nmo.13292.
[17]FAN W,FEI G,LI X,et al.Sera with anti-enteric neuronal antibodies from patients with irritable bowel syndrome promote apoptosis in myenteric neurons of guinea pigs and human SH-Sy5Y cells[J].Neurogastroenterol Motil,2018,30(12):e13457.DOI:10.1111/nmo.13457.
[18]OHLSSON B,GUSTAFSSON R,SWAHN F,et al.Endoscopic fullthickness biopsy,a novel method in the work up of complicated abdominal symptoms[J].Therap Adv Gastroenterol,2017,11:1756283X17730747.DOI:10.1177/1756283X17730747.
[19]OHLSSON B,GUSTAFSSON R J,TOTH E,et al.Endoscopic versus laparoscopic full-thickness biopsy in the pathological evaluation of the enteric nervous system[J].Case Rep Gastroenterol,2018,12(1):32-40.DOI:10.1159/000486390.
[2]陈玲玲,文平,费贵军,等.腹泻型肠易激综合征患者血清抗肠神经元抗体免疫反应特点及其临床意义[J].基础医学与临床,2017,37(2):156-161.DOI:10.3969/j.issn.1001-6325.2017.02.007.CHEN L L,WEN P,FEI G J,et al.Characteristics and clinical significance of sera anti-enteric neuronal antibodies in patients with irritable bowel syndrome with diarrhea[J].Basic&Clinical Medicine,2017,37(2):156-161.DOI:10.3969/j.issn.1001-6325.2017.02.007.
[3]李珊,费贵军,方秀才,等.腹泻型肠易激综合征模型大鼠结肠黏膜下神经丛神经元的改变[J].中华消化杂志,2010,30(4):250-253.DOI:10.3760/cma.j.issn.0254-1432.2010.04.008.LI S,FEI G J,FANG X C,et al.The morphological changes of colonic submucosal plexus in rats of irritable bowel syndrome with diarrhea[J].Chin J Dig,2010,30(4):250-253.DOI:10.3760/cma.j.issn.0254-1432.2010.04.008.
[4]MANN P T,SOUTHWELL B R,DING Y Q,et al.Localisation of neurokinin 3(NK3)receptor immunoreactivity in the rat gastrointestinal tract[J].Cell Tissue Res,1997,289(1):1-9.
[5]吕红,钱家鸣,金光亮,等.肠易激综合征慢急性联合应激动物模型的建立及其感觉、动力和心理行为的评价[J].中华内科杂志,2009,48(12):1035-1039.DOI:10.3760/cma.j.issn.0578-1426.2009.12.015.LYU H,QIAN J M,JIN G L,et al.The establishment of an animal model of gut-brain interaction in irritable bowel syndrome for the evaluation of visceral sensation,motility and psychological behavior[J].Chin J Intern Med,2009,48(12):1035-1039.DOI:10.3760/cma.j.issn.0578-1426.2009.12.015.
[6]彭丽华,杨云生,孙刚,等.便秘型肠易激综合征新概念模型的建立[J].世界华人消化杂志,2004,12(1):112-116.DOI:10.3969/j.issn.1009-3079.2004.01.029.PENG L H,YANG Y S,SUN G,et al.A new model of constipation-predominant irritable bowel syndrome in rats[J].World Chin J Digestol,2004,12(1):112-116.DOI:10.3969/j.issn.1009-3079.2004.01.029.
[7]杨希林,方秀才,刘晓红,等.便秘型肠易激综合征大鼠胃排空及胃内肌间神经丛神经元的变化[J].基础医学与临床,2011,31(12):1301-1304.YANG X L,FANG X C,LIU X H,et al.The changes of gastric emptying and gastric myenteric neurons in rats with IBS complicated with constipation[J].Basic&Clinical Medicine,2011,31(12):1301-1304.
[8]孙巧丽,袁建业,谢建群.肠易激综合征动物模型的研究现状和展望[J].中华消化杂志,2012,32(2):140-142.DOI:10.3760/cma.j.issn.0254-1432.2012.02.020.
[9]SCHWALLER B.Calretinin:from a“simple”Ca(2+)buffer to a multifunctional protein implicated in many biological processes[J].Front Neuroanat,2014,8:3.DOI:10.3389/fnana.2014.00003.
[10]ORD EZ N G.Value of calretinin immunostaining in diagnostic pathology:a review and update[J].Appl Immunohistochem Mol Morphol,2014,22(6):401-415.DOI:10.1097/PAI.0b013e31829b6fbd.
[11]GAO N,LUO J,URAY K,et al.CaMKⅡis essential for the function of the enteric nervous system[J].PLo S One,2012,7(8):e44426.DOI:10.1371/journal.pone.0044426.
[12]MITSUI R.Immunohistochemical characteristics of submucosal Dogiel typeⅡneurons in rat colon[J].Cell Tissue Res,2010,340(2):257-265.DOI:10.1007/s00441-010-0954-z.
[13]BEUSCHER N,JABARI S,STREHL J,et al.What neurons hide behind calretinin immunoreactivity in the human gut?[J].Histochem Cell Biol,2014,141(4):393-405.DOI:10.1007/s00418-013-1163-0.
[14]ZETZMANN K,STREHL J,GEPPERT C,et al.Calbindin D28kimmunoreactivity in human enteric neurons[J].Int J Mol Sci,2018,19(1):pii:E194.DOI:10.3390/ijms19010194.
[15]JAHNG J,KIM Y S.Irritable bowel syndrome:is it really a functional disorder?A new perspective on alteration of enteric nervous system[J].J Neurogastroenterol Motil,2016,22(2):163-165.DOI:10.5056/jnm16043.
[16]LTT A,MICHEL K,KRGER D,et al.High prevalence and functional effects of serum antineuronal antibodies in patients with gastrointestinal disorders[J].Neurogastroenterol Motil,2018,30(6):e13292.DOI:10.1111/nmo.13292.
[17]FAN W,FEI G,LI X,et al.Sera with anti-enteric neuronal antibodies from patients with irritable bowel syndrome promote apoptosis in myenteric neurons of guinea pigs and human SH-Sy5Y cells[J].Neurogastroenterol Motil,2018,30(12):e13457.DOI:10.1111/nmo.13457.
[18]OHLSSON B,GUSTAFSSON R,SWAHN F,et al.Endoscopic fullthickness biopsy,a novel method in the work up of complicated abdominal symptoms[J].Therap Adv Gastroenterol,2017,11:1756283X17730747.DOI:10.1177/1756283X17730747.
[19]OHLSSON B,GUSTAFSSON R J,TOTH E,et al.Endoscopic versus laparoscopic full-thickness biopsy in the pathological evaluation of the enteric nervous system[J].Case Rep Gastroenterol,2018,12(1):32-40.DOI:10.1159/000486390.