肠神经胶质细胞形态学研究方法进展
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摘要
肠神经胶质细胞(enteric glial cells, EGCs)是肠神经系统的重要组成部分,其数量约是肠神经元数量的4倍.在形态和功能上与中枢神经系统的星形胶质细胞相似.在营养、支持肠神经、维持胃肠道稳态、调节胃肠道功能等方面发挥着重要作用.由于其在肠道中发挥的作用复杂而多样,成为研究的焦点.对其形态学的观察和研究是非常重要的.EGCs发现至今,针对其发挥功能的作用机制已得到深入研究,相应的研究方法也在不断的进步与改进中.本文主要介绍EGCs在胃肠道生理及病理状态下形态学特点及目前形态学研究方法,以期对未来研究EGCs提供实验方法的借鉴及参考意义,推进这一领域的发展.
As an important part of the intestinal nervous system, enteric glial cells are about four times as many as intestinal neurons.Furthermore, a large population of astrocyte-like glial cells populate gut muscle layers and the intestinal mucosa, and mounting new evidence points toward enteric glial cells as an active participant in gut pathology.They are similar in morphology and function to the astrocytes of the central nervous system and play an important role in nutrition, supporting gastrointestinal nerve, maintaining gastrointestinal homeostasis, and regulating gastrointestinal function.Because of their complex and diverse roles in the intestinal tract, they have become the focus of research.As the study of their functional mechanism has been extensively deepened, the research methods for intestinal glial cells are also on constant progress and improvement, especially in studying their morphology.This paper mainly introduces the morphological characteristics of enteric glial cells under the conditions of gastrointestinal physiology and pathology, so as to provide a reference for the future study of enteric glial cells and promote the development of this field.
As an important part of the intestinal nervous system, enteric glial cells are about four times as many as intestinal neurons.Furthermore, a large population of astrocyte-like glial cells populate gut muscle layers and the intestinal mucosa, and mounting new evidence points toward enteric glial cells as an active participant in gut pathology.They are similar in morphology and function to the astrocytes of the central nervous system and play an important role in nutrition, supporting gastrointestinal nerve, maintaining gastrointestinal homeostasis, and regulating gastrointestinal function.Because of their complex and diverse roles in the intestinal tract, they have become the focus of research.As the study of their functional mechanism has been extensively deepened, the research methods for intestinal glial cells are also on constant progress and improvement, especially in studying their morphology.This paper mainly introduces the morphological characteristics of enteric glial cells under the conditions of gastrointestinal physiology and pathology, so as to provide a reference for the future study of enteric glial cells and promote the development of this field.
引文
1 Gulbransen BD,Sharkey KA.Novel functional roles for enteric glia in the gastrointestinal tract.Nat Rev Gastroenterol Hepatol 2012;9:625-632[PMID:22890111 DOI:10.1038/nrgastro.2012.138]
2 Gabella G.Fine structure of the myenteric plexus in the guineapig ileum.J Anat 1972;111:69-97[PMID:4335909]
3 Rühl A.Glial cells in the gut.Neurogastroenterol Motil 2005;17:777-790[PMID:16336493DOI:10.1111/j.1365-2982.2005.00687.x]
4 Hanani M,Reichenbach A.Morphology of horseradish peroxidase(HRP)-injected glial cells in the myenteric plexus of the guinea-pig.Cell Tissue Res 1994;278:153-160[PMID:7954696 DOI:10.1007/bf00305787]
5 Bohórquez DV,Samsa LA,Roholt A,Medicetty S,Chandra R,Liddle RA.An enteroendocrine cell-enteric glia connection revealed by 3D electron microscopy.PLo S One 2014;9:e89881[PMID:24587096 DOI:10.1371/journal.pone.0089881]
6 De Giorgio R,Giancola F,Boschetti E,Abdo H,Lardeux B,Neunlist M.Enteric glia and neuroprotection:basic and clinical aspects.Am J Physiol Gastrointest Liver Physiol 2012;303:G887-G893[PMID:22878122 DOI:10.1152/ajpgi.00096.2012]
7 Coelho-Aguiar Jde M,Bon-Frauches AC,Gomes AL,Veríssimo CP,Aguiar DP,Matias D,Thomasi BB,Gomes AS,Brito GA,Moura-Neto V.The enteric glia:identity and functions.Glia2015;63:921-935[PMID:25703790 DOI:10.1002/glia.22795]
8 Liu YA,Chung YC,Pan ST,Shen MY,Hou YC,Peng SJ,Pasricha PJ,Tang SC.3-D imaging,illustration,and quantitation of enteric glial network in transparent human colon mucosa.Neurogastroenterol Motil 2013;25:e324-e338[PMID:23495930 DOI:10.1111/nmo.12115]
9 Savidge TC,Sofroniew MV,Neunlist M.Starring roles for astroglia in barrier pathologies of gut and brain.Lab Invest 2007;87:731-736[PMID:17607301 DOI:10.1038/labinvest.3700600]
10 Neunlist M,Van Landeghem L,MahéMM,Derkinderen P,des Varannes SB,Rolli-Derkinderen M.The digestive neuronalglial-epithelial unit:a new actor in gut health and disease.Nat Rev Gastroenterol Hepatol 2013;10:90-100[PMID:23165236 DOI:10.1038/nrgastro.2012.221]
11 Boesmans W,Lasrado R,Vanden Berghe P,Pachnis V.Heterogeneity and phenotypic plasticity of glial cells in the mammalian enteric nervous system.Glia 2015;63:229-241[PMID:25161129 DOI:10.1002/glia.22746]
12 Sharkey KA.Emerging roles for enteric glia in gastrointestinal disorders.J Clin Invest 2015;125:918-925[PMID:25689252DOI:10.1172/JCI76303]
13 Fujikawa Y,Tominaga K,Tanaka F,Tanigawa T,Watanabe T,Fujiwara Y,Arakawa T.Enteric glial cells are associated with stress-induced colonic hyper-contraction in maternally separated rats.Neurogastroenterol Motil 2015;27:1010-1023[PMID:25960044 DOI:10.1111/nmo.12577]
14 Grubi?i?V,Gulbransen BD.Enteric glia:the most alimentary of all glia.J Physiol 2017;595:557-570[PMID:27106597 DOI:10.1113/JP271021]
15赵增凯.电针足三里对失血性休克大鼠肠屏障和肠胶质细胞保护作用的研究.2015;15:136-139[DOI:10.3969/j.issn.1672-8521.2014.03.002]
16 Hu S,Zhao ZK,Liu R,Wang HB,Gu CY,Luo HM,Wang H,Du MH,Lv Y,Shi X.Electroacupuncture activates enteric glial cells and protects the gut barrier in hemorrhaged rats.World J Gastroenterol 2015;21:1468-1478[PMID:25663766 DOI:10.3748/wjg.v21.i5.1468]
17王鹏.肠易激综合征肠上皮-肠胶质细胞网络调控异常的分子机制研究.山东大学,2015
18刘丽莎,张微,赵敏,毛廷丽,李思宇,李瑛,钟振东.针刺大肠俞募穴对慢传输便秘小鼠结肠组织形态学的影响.中华中医药杂志2016;31:255-258
19 Mendes CE,Palombit K,Vieira C,Silva I,Correia-de-SáP,Castelucci P.The Effect of Ischemia and Reperfusion on Enteric Glial Cells and Contractile Activity in the Ileum.Dig Dis Sci 2015;60:2677-2689[PMID:25917048 DOI:10.1007/s10620-015-3663-3]
20 Bassotti G,Villanacci V,Antonelli E,Morelli A,Salerni B.Enteric glial cells:new players in gastrointestinal motility.Lab Invest 2007;87:628-632[PMID:17483847 DOI:10.1038/labinvest.3700564]
21 Coons AH.The development of immunohistochemistry.Ann N Y Acad Sci 1971;177:5-9[PMID:44005566 DOI:10.1111/j.17496632.1971.tb35025.x]
22 Rossi S,Laurino L,Furlanetto A,Chinellato S,Orvieto E,Canal F,Facchetti F,Dei Tos AP.Rabbit monoclonal antibodies:a comparative study between a novel category of immunoreagents and the corresponding mouse monoclonal antibodies.Am J Clin Pathol 2005;124:295-302[PMID:16040303DOI:10.1309/NR8H-N08G-DPVE-MU08]
23 Ward JM,Rehg JE.Rodent immunohistochemistry:pitfalls and troubleshooting.Vet Pathol 2014;51:88-101[PMID:24078006DOI:10.1177/0300985813503571]
24 Cirillo C,Sarnelli G,Esposito G,Turco F,Steardo L,Cuomo R.S100B protein in the gut:the evidence for enteroglialsustained intestinal inflammation.World J Gastroenterol 2011;17:1261-1266[PMID:21455324 DOI:10.3748/wjg.v17.i10.1261]
25 Hoff S,Zeller F,von Weyhern CW,Wegner M,Schemann M,Michel K,Rühl A.Quantitative assessment of glial cells in the human and guinea pig enteric nervous system with an antiSox8/9/10 antibody.J Comp Neurol 2008;509:356-371[PMID:18512230 DOI:10.1002/cne.21769]
26 Bassotti G,Villanacci V,Maurer CA,Fisogni S,Di Fabio F,Cadei M,Morelli A,Panagiotis T,Cathomas G,Salerni B.The role of glial cells and apoptosis of enteric neurones in the neuropathology of intractable slow transit constipation.Gut2006;55:41-46[PMID:16041063 DOI:10.1136/gut.2005.073197]
27杨敏,田志华,马博,贾凯英,张鸿丽,富晶,綦雯雯,赵威.多重间接免疫荧光技术的建立及其在肝癌样本多种抗原检测中的应用.细胞与分子免疫学杂志2018;34:97-104[DOI:10.13423/j.cnki.cjcmi.008522]
28 Le Berre-Scoul C,Chevalier J,Oleynikova E,Cossais F,Talon S,Neunlist M,Boudin H.A novel enteric neuron-glia coculture system reveals the role of glia in neuronal development.JPhysiol 2017;595:583-598[PMID:27436013 DOI:10.1113/JP271989]
29 Cirillo C,Bessissow T,Desmet AS,Vanheel H,Tack J,Vanden Berghe P.Evidence for neuronal and structural changes in submucous ganglia of patients with functional dyspepsia.Am J Gastroenterol 2015;110:1205-1215[PMID:26077177 DOI:10.1038/ajg.2015.158]
30白艳华,张军.大鼠胃肠道肌间神经丛铺片的制备和染色法的研究.第四军医大学学报2009;30:1950-1952
31 Pearson GT.Structural organization and neuropeptide distributions in the equine enteric nervous system:an immunohistochemical study using whole-mount preparations from the small intestine.Cell Tissue Res 1994;276:523-534[PMID:7520362 DOI:10.1007/bf00343949]
32纪托,薛红,朱进霞.显示肠肌间神经丛的全层铺片技术.首都医科大学学报2009;30:256-258[DOI:10.3969/j.issn.1006-7795.2009.02.031]
33张国权,孙海梅,夏白娟,李小双,康倩,江涛.全层铺片技术研究消化道神经丛和Cajal间质细胞.解剖学研2013;35:76-78[DOI:10.3969/j.issn.167-0770.2013.01.022]
34周德山.电镜技术在医学生物学领域应用体会及历史回顾.中国解剖学会.解剖学杂志-中国解剖学会2002年年会文摘汇编.中国解剖学会2002;2
35 Mitchison HM,Shoemark A.Motile cilia defects in diseases other than primary ciliary dyskinesia:The contemporary diagnostic and research role for transmission electron microscopy.Ultrastruct Pathol 2017;41:415-427[PMID:28925789 DOI:10.1080/01913123.2017.1370050]
36王萌,李婷,董红燕.透射电镜制样过程中漂洗时间对神经组织超微结构的影响.神经解剖学杂志2018;34:515-518[DOI:10.16557/j.cnki.1000-7547.2018.04.017]
2 Gabella G.Fine structure of the myenteric plexus in the guineapig ileum.J Anat 1972;111:69-97[PMID:4335909]
3 Rühl A.Glial cells in the gut.Neurogastroenterol Motil 2005;17:777-790[PMID:16336493DOI:10.1111/j.1365-2982.2005.00687.x]
4 Hanani M,Reichenbach A.Morphology of horseradish peroxidase(HRP)-injected glial cells in the myenteric plexus of the guinea-pig.Cell Tissue Res 1994;278:153-160[PMID:7954696 DOI:10.1007/bf00305787]
5 Bohórquez DV,Samsa LA,Roholt A,Medicetty S,Chandra R,Liddle RA.An enteroendocrine cell-enteric glia connection revealed by 3D electron microscopy.PLo S One 2014;9:e89881[PMID:24587096 DOI:10.1371/journal.pone.0089881]
6 De Giorgio R,Giancola F,Boschetti E,Abdo H,Lardeux B,Neunlist M.Enteric glia and neuroprotection:basic and clinical aspects.Am J Physiol Gastrointest Liver Physiol 2012;303:G887-G893[PMID:22878122 DOI:10.1152/ajpgi.00096.2012]
7 Coelho-Aguiar Jde M,Bon-Frauches AC,Gomes AL,Veríssimo CP,Aguiar DP,Matias D,Thomasi BB,Gomes AS,Brito GA,Moura-Neto V.The enteric glia:identity and functions.Glia2015;63:921-935[PMID:25703790 DOI:10.1002/glia.22795]
8 Liu YA,Chung YC,Pan ST,Shen MY,Hou YC,Peng SJ,Pasricha PJ,Tang SC.3-D imaging,illustration,and quantitation of enteric glial network in transparent human colon mucosa.Neurogastroenterol Motil 2013;25:e324-e338[PMID:23495930 DOI:10.1111/nmo.12115]
9 Savidge TC,Sofroniew MV,Neunlist M.Starring roles for astroglia in barrier pathologies of gut and brain.Lab Invest 2007;87:731-736[PMID:17607301 DOI:10.1038/labinvest.3700600]
10 Neunlist M,Van Landeghem L,MahéMM,Derkinderen P,des Varannes SB,Rolli-Derkinderen M.The digestive neuronalglial-epithelial unit:a new actor in gut health and disease.Nat Rev Gastroenterol Hepatol 2013;10:90-100[PMID:23165236 DOI:10.1038/nrgastro.2012.221]
11 Boesmans W,Lasrado R,Vanden Berghe P,Pachnis V.Heterogeneity and phenotypic plasticity of glial cells in the mammalian enteric nervous system.Glia 2015;63:229-241[PMID:25161129 DOI:10.1002/glia.22746]
12 Sharkey KA.Emerging roles for enteric glia in gastrointestinal disorders.J Clin Invest 2015;125:918-925[PMID:25689252DOI:10.1172/JCI76303]
13 Fujikawa Y,Tominaga K,Tanaka F,Tanigawa T,Watanabe T,Fujiwara Y,Arakawa T.Enteric glial cells are associated with stress-induced colonic hyper-contraction in maternally separated rats.Neurogastroenterol Motil 2015;27:1010-1023[PMID:25960044 DOI:10.1111/nmo.12577]
14 Grubi?i?V,Gulbransen BD.Enteric glia:the most alimentary of all glia.J Physiol 2017;595:557-570[PMID:27106597 DOI:10.1113/JP271021]
15赵增凯.电针足三里对失血性休克大鼠肠屏障和肠胶质细胞保护作用的研究.2015;15:136-139[DOI:10.3969/j.issn.1672-8521.2014.03.002]
16 Hu S,Zhao ZK,Liu R,Wang HB,Gu CY,Luo HM,Wang H,Du MH,Lv Y,Shi X.Electroacupuncture activates enteric glial cells and protects the gut barrier in hemorrhaged rats.World J Gastroenterol 2015;21:1468-1478[PMID:25663766 DOI:10.3748/wjg.v21.i5.1468]
17王鹏.肠易激综合征肠上皮-肠胶质细胞网络调控异常的分子机制研究.山东大学,2015
18刘丽莎,张微,赵敏,毛廷丽,李思宇,李瑛,钟振东.针刺大肠俞募穴对慢传输便秘小鼠结肠组织形态学的影响.中华中医药杂志2016;31:255-258
19 Mendes CE,Palombit K,Vieira C,Silva I,Correia-de-SáP,Castelucci P.The Effect of Ischemia and Reperfusion on Enteric Glial Cells and Contractile Activity in the Ileum.Dig Dis Sci 2015;60:2677-2689[PMID:25917048 DOI:10.1007/s10620-015-3663-3]
20 Bassotti G,Villanacci V,Antonelli E,Morelli A,Salerni B.Enteric glial cells:new players in gastrointestinal motility.Lab Invest 2007;87:628-632[PMID:17483847 DOI:10.1038/labinvest.3700564]
21 Coons AH.The development of immunohistochemistry.Ann N Y Acad Sci 1971;177:5-9[PMID:44005566 DOI:10.1111/j.17496632.1971.tb35025.x]
22 Rossi S,Laurino L,Furlanetto A,Chinellato S,Orvieto E,Canal F,Facchetti F,Dei Tos AP.Rabbit monoclonal antibodies:a comparative study between a novel category of immunoreagents and the corresponding mouse monoclonal antibodies.Am J Clin Pathol 2005;124:295-302[PMID:16040303DOI:10.1309/NR8H-N08G-DPVE-MU08]
23 Ward JM,Rehg JE.Rodent immunohistochemistry:pitfalls and troubleshooting.Vet Pathol 2014;51:88-101[PMID:24078006DOI:10.1177/0300985813503571]
24 Cirillo C,Sarnelli G,Esposito G,Turco F,Steardo L,Cuomo R.S100B protein in the gut:the evidence for enteroglialsustained intestinal inflammation.World J Gastroenterol 2011;17:1261-1266[PMID:21455324 DOI:10.3748/wjg.v17.i10.1261]
25 Hoff S,Zeller F,von Weyhern CW,Wegner M,Schemann M,Michel K,Rühl A.Quantitative assessment of glial cells in the human and guinea pig enteric nervous system with an antiSox8/9/10 antibody.J Comp Neurol 2008;509:356-371[PMID:18512230 DOI:10.1002/cne.21769]
26 Bassotti G,Villanacci V,Maurer CA,Fisogni S,Di Fabio F,Cadei M,Morelli A,Panagiotis T,Cathomas G,Salerni B.The role of glial cells and apoptosis of enteric neurones in the neuropathology of intractable slow transit constipation.Gut2006;55:41-46[PMID:16041063 DOI:10.1136/gut.2005.073197]
27杨敏,田志华,马博,贾凯英,张鸿丽,富晶,綦雯雯,赵威.多重间接免疫荧光技术的建立及其在肝癌样本多种抗原检测中的应用.细胞与分子免疫学杂志2018;34:97-104[DOI:10.13423/j.cnki.cjcmi.008522]
28 Le Berre-Scoul C,Chevalier J,Oleynikova E,Cossais F,Talon S,Neunlist M,Boudin H.A novel enteric neuron-glia coculture system reveals the role of glia in neuronal development.JPhysiol 2017;595:583-598[PMID:27436013 DOI:10.1113/JP271989]
29 Cirillo C,Bessissow T,Desmet AS,Vanheel H,Tack J,Vanden Berghe P.Evidence for neuronal and structural changes in submucous ganglia of patients with functional dyspepsia.Am J Gastroenterol 2015;110:1205-1215[PMID:26077177 DOI:10.1038/ajg.2015.158]
30白艳华,张军.大鼠胃肠道肌间神经丛铺片的制备和染色法的研究.第四军医大学学报2009;30:1950-1952
31 Pearson GT.Structural organization and neuropeptide distributions in the equine enteric nervous system:an immunohistochemical study using whole-mount preparations from the small intestine.Cell Tissue Res 1994;276:523-534[PMID:7520362 DOI:10.1007/bf00343949]
32纪托,薛红,朱进霞.显示肠肌间神经丛的全层铺片技术.首都医科大学学报2009;30:256-258[DOI:10.3969/j.issn.1006-7795.2009.02.031]
33张国权,孙海梅,夏白娟,李小双,康倩,江涛.全层铺片技术研究消化道神经丛和Cajal间质细胞.解剖学研2013;35:76-78[DOI:10.3969/j.issn.167-0770.2013.01.022]
34周德山.电镜技术在医学生物学领域应用体会及历史回顾.中国解剖学会.解剖学杂志-中国解剖学会2002年年会文摘汇编.中国解剖学会2002;2
35 Mitchison HM,Shoemark A.Motile cilia defects in diseases other than primary ciliary dyskinesia:The contemporary diagnostic and research role for transmission electron microscopy.Ultrastruct Pathol 2017;41:415-427[PMID:28925789 DOI:10.1080/01913123.2017.1370050]
36王萌,李婷,董红燕.透射电镜制样过程中漂洗时间对神经组织超微结构的影响.神经解剖学杂志2018;34:515-518[DOI:10.16557/j.cnki.1000-7547.2018.04.017]