人胎小肠肥大细胞的发生与神经肽分布和发育的研究
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摘要
目的：①观察胎儿小肠(十二指肠、空肠和回肠)发育的组织形态变化。②研究胎儿小肠壁肥大细胞(mast cells MC)的发育规律和异质性。③研究人胎儿小肠壁P物质(SP)和降钙素基因相关肽(CGRP)-IR肽能神经的分布和发育规律。④探讨胎儿小肠MC与SP-IR、CGRP-IR肽能神经的关系。方法：①苏木精伊红(HE)染色；②甲苯胺蓝(TB)特殊染色；③免疫组织化学ABC法。结果：①小肠组织的分化是从头端向尾端进行。第12周时，十二指肠形成较发达的叶状肠绒毛，空肠指状绒毛较明显，回肠为低矮的锥体形突起；潘氏细胞显现时间十二指肠、空肠、回肠分别为15周、16周、17周。第15周时，可见十二指肠中的肠腺延伸入粘膜下层形成粘液性细胞组成的十二指肠腺。第21周时，粘膜肌层出现，小肠壁四层分界渐清楚。②幼稚的MC颗粒用TB染色呈淡紫色，分化成熟的MC颗粒渐多，染成深紫色。第16周时，在小肠粘膜下层、肌层间结缔组织中偶见结缔组织MC，而粘膜MC出现在18周，结缔组织MC和粘膜MC有异质性。两型MC中某些自22周开始呈活化状态，有些MC表面界限不清，有脱颗粒现象，胎儿MC与血管、神经节等密切接触，MC可能与调节小肠局部血流及感觉等功能密切相关。③第14周时，胎儿小肠壁粘膜下层、肌层间结缔组织中偶见SP能、CGRP能神经纤维及神经元免疫反应产物，反应渐强，神经元从浅棕色到深棕色；神经纤维呈串珠状或点线状。从SP、CGRP免疫组织化学相邻切片上看，两者部分存在共存现象。④从小肠壁TB特殊染色MC和SP、CGRP免疫组织化学相邻切片上看，部分MC内存在SP、CGRP免疫反应阳性物质。结论：①在胎儿发育过程中，小肠组织形态出现明显的变化，其分化由十二指肠向空肠、回肠依次进行。②胎儿小肠MC有粘膜MC和结缔组织MC，两者存在异质性。③人胎小肠壁存在SP-IR、CGRP-IR肽能神经，粘膜下和肌间神经丛存在
安徽医科大学硕士学位论文
SP．IR、CGRP．皿肽能神经元和神经纤维，它们在发生、分布有差异。④胎儿小
肠某些MC内存在SPIR、CGny－IR肽能活性物质，MC可能与局部血液循环和
感觉等有关
Objectives(1)To observe the histomorphologic development of human fetus small intestine (duodenum-, jejunum and ileum). (2) To study the developmental regularities and heterogeneity of mast cells in fetus small intestine. (3) To study the distribution and developmental regularities of Substance P (SP) and Calcitonin Gene-Related Peptide (CGRP) - IR peptidergic nerves in fetus small intestine. ?To approach the relationship of the mast cells(MC) and SP- IR and CGRP- IR peptidergic nerves in fetus small intestine Methods (1)Hematoxylin-Eosin (HE) staining. (2)Toluidine blue (TB) staining. (3) Immunohistochemical ABC method. Results. (1) The histological differentiations in small intestine were proceed from the beginning to the end .At 12nd week, the fairly developed lobe-shape intestinal villus were formed hi duodenum, Digital-shape villus had also seen in jejunum, but lower-conus body configur appeared in ileum. At 15th week, intestinal gland in duodenum extend into submucosa to form mucous cell's duodenal gland. At 21st week, mucosa muscularis appeared
gradually ,there were four layers in the wall of small intestine. (2)The granular of the immaure MC were pale violet and the granular of the mature MC were strong violet in color by TB staining. At 16 th week, connective tissue MC (CTMC) appeared occasionally in connective tissue of submucosa and muscular layer in small intestine. But the time of MC in mucosa (MMC) appeared at 18th week, So MC has heterogeneity. At 22nd week, the two types of MC were both active. They were distributed in the surrounding blood vessels and ganglions, etc. The verge of some MC were unclear, they were degranular phenomena. MC may play an important part in regulating blood circulation and sensation. (3)At 14th week, The SP-IR and CGRP-IR nerve fibers and cells appeared occasionally in the myenteric and submucous plexuses in small intestine, and the responds were gradually strong. Neuron showed from light to deep brown, and nerve fibers were presented as varicose and liner profiles. On the corresponding site of serial sections of
SP-IR , CGRP-IR immunohistochemistry, some of them were coexisted in one nerve fiber and cells. (4)On the corresponding site of serial sections of TB special staining and SP- DR^ CGRP- IR immunohistochemistry, some of MC showed SP-IR, CGRP-IR immunoreactivity. Conclusion (1)The histologic form of small intestine appeared obviousely changing in developing fetus, Its differentiation proceeds successively from duodenum to jejunum and ileum. (2)There were two kinds of MC--mucosa MC and connective tissue MC, which showed heterogeneity.(3)The SP-IR and CGRP-IR peptidergic nerves were seen in fetus small intestine. The SP-IR and CGRP-IR nerve fibers and cells appeared in the myenteric and submucous plexuses, but their development and distributions were different. (4) Some of MC in fetus small intestine showed SP^ CGRP- immunoreactivity. MC may play an important part in regulating blood circulation and sensation.
安徽医科大学硕士学位论文
SP．IR、CGRP．皿肽能神经元和神经纤维，它们在发生、分布有差异。④胎儿小
肠某些MC内存在SPIR、CGny－IR肽能活性物质，MC可能与局部血液循环和
感觉等有关
Objectives(1)To observe the histomorphologic development of human fetus small intestine (duodenum-, jejunum and ileum). (2) To study the developmental regularities and heterogeneity of mast cells in fetus small intestine. (3) To study the distribution and developmental regularities of Substance P (SP) and Calcitonin Gene-Related Peptide (CGRP) - IR peptidergic nerves in fetus small intestine. ?To approach the relationship of the mast cells(MC) and SP- IR and CGRP- IR peptidergic nerves in fetus small intestine Methods (1)Hematoxylin-Eosin (HE) staining. (2)Toluidine blue (TB) staining. (3) Immunohistochemical ABC method. Results. (1) The histological differentiations in small intestine were proceed from the beginning to the end .At 12nd week, the fairly developed lobe-shape intestinal villus were formed hi duodenum, Digital-shape villus had also seen in jejunum, but lower-conus body configur appeared in ileum. At 15th week, intestinal gland in duodenum extend into submucosa to form mucous cell's duodenal gland. At 21st week, mucosa muscularis appeared
gradually ,there were four layers in the wall of small intestine. (2)The granular of the immaure MC were pale violet and the granular of the mature MC were strong violet in color by TB staining. At 16 th week, connective tissue MC (CTMC) appeared occasionally in connective tissue of submucosa and muscular layer in small intestine. But the time of MC in mucosa (MMC) appeared at 18th week, So MC has heterogeneity. At 22nd week, the two types of MC were both active. They were distributed in the surrounding blood vessels and ganglions, etc. The verge of some MC were unclear, they were degranular phenomena. MC may play an important part in regulating blood circulation and sensation. (3)At 14th week, The SP-IR and CGRP-IR nerve fibers and cells appeared occasionally in the myenteric and submucous plexuses in small intestine, and the responds were gradually strong. Neuron showed from light to deep brown, and nerve fibers were presented as varicose and liner profiles. On the corresponding site of serial sections of
SP-IR , CGRP-IR immunohistochemistry, some of them were coexisted in one nerve fiber and cells. (4)On the corresponding site of serial sections of TB special staining and SP- DR^ CGRP- IR immunohistochemistry, some of MC showed SP-IR, CGRP-IR immunoreactivity. Conclusion (1)The histologic form of small intestine appeared obviousely changing in developing fetus, Its differentiation proceeds successively from duodenum to jejunum and ileum. (2)There were two kinds of MC--mucosa MC and connective tissue MC, which showed heterogeneity.(3)The SP-IR and CGRP-IR peptidergic nerves were seen in fetus small intestine. The SP-IR and CGRP-IR nerve fibers and cells appeared in the myenteric and submucous plexuses, but their development and distributions were different. (4) Some of MC in fetus small intestine showed SP^ CGRP- immunoreactivity. MC may play an important part in regulating blood circulation and sensation.
引文
1 Enerbaek L. Mast cells in rat gastrointestinal mucosa, Acta Path. Microbiol. Scandinav, 1966, 66:303～306
2 Irani A, Schechter M, Craig S, et al. Two types of human mast cells that have distinct neutral protease composition. Proc Natl Acad Sci USA, 1986, 83: 4464～4468
3 董增军．肥大细胞亚群．国外医学免疫学分册，1990，13：185-186
4 Kitamura Y. Development of mast cell from grafted bone marrow cells in irradiated mice. Nature, 1977, 268:442～446
5 Sonda BT. Prolifetation of peritoneal mast cells in the skin of W/W~v mice that genetically lack mast cells. J Exp. Med. 1984, 160:138～140
6 Steven RL, Rothemberg ME, Levi-Schaffer F, et al. Ontogeny of in vitro differentiated mouse mast cells. Fed Proc, 1987, 46:1915～1918
7 Sonoda S, Sonoda T, Nakano T, et al. Development of mucosa mast cells after injection of a single connective tissue-type mast cell in the stomach mucosa of genetically mast cell deficient W/W~v mice. J Immunol, 1986, 137(4): 1319～1322
8 杨美林，高崇，乔从进，等，肥大细胞与成纤维细胞功能关系的光镜、电镜研究．解剖学杂志，1992，15(3)：289～291
9 Marks RM, Roche WR, Czerniecki M, et al. Mast cell granules cause proliferation of human microvascular endothelial cells. Lab Invest, 1986, 55:289～291
10 Von Euler US, Gaddum JH. An unidentified depressor substance in certain tissue extracts. J Physiol. 1931, 72: 74～78
11 邹仲之，陈东，尹昕．消化管．见：成令忠主编．组织学(第二版)．北京，人民卫生出版社．1993，1126～1139
12 朱文玉主编：神经和胃肠道肽类的关系．生理学进展．1980，11：259～261
13 李在琉．胃肠运动的神经控制．见：周吕主编．胃肠生理学——基础与临床．北京，科学出版社．1998，3(2)，363～379
14 Stead RH. Mast cells are closely apposed to nerves in the human gastrointestinal mucosa. Gastroenterology, 1989, 97:575～578
15 王俊明，张得真．P物质和血管活性肠肽免疫反应在人皮肤内经络线上肥大细胞颗粒内的定位研究．解剖学杂志．1986，9(增刊)：372～376
16 贾雪梅、贾友苏，齐威琴，等．P物质、血管活性肠肽和神经肽Y在大鼠舌肥大细胞的定位研究，解剖学杂志，1998，21(3)：242～246
17 Blennerhassett MG. Mast cells are closely apposed to nerves in the mice gastrointestinal mucosa. Cell Tissue Res, 1991, 265(1): 121～128
18 周金黄．神经免疫调节的进展．生理科学进展．1987，18(3)：119～122
19 Wingren U. Mucosal mast cells of the rat intestine: a rervaluation of fixation and staining properties with special reference to protein blocking and solubility of the granular glycosaminoglycan. Histochemical, 1983, 15: 571～572
20 何素云．消化和呼吸系统的发生．见：刘斌，高英茂主编．人体胚胎学(第二版)．北京：人民卫生出版社，1996：253～255
21 Lev R, Siegel HI, Bantman J. Histochemical studies of developing human fetal small intestine. Histochemistry, 1972, 29(2): 103～119
22 Combs JW. Differentiation and proliferation of embryonic mast cells of the rat. J cell Biol. 1965, 25:577～580
23 翟力平，杨美林．发育中人胎儿肥大细胞的组织化学研究．中国组织化学与细胞化学杂志，1995，4(2)，139～142
24 周开渠，彭庆廉．固有结缔组织．见：成令忠主编．组织学，第二版，北京：人民卫生出版社，1993，234～243
25 Craig S, Schechter M, Schwants B, Ultrastructural analysis of maturing human T and TC mast cells identified by immunoelectron. Lab Invest, 1988, 58: 682～688
26 翟力平，安巍，杨美林，等．胎儿消化器官发育中的肥大细胞超微结构特点．解剖学杂志，1997，20(1)，57～60
27 Dralce-Lee AB, Price J. Ultrastructure of mast cells in normal subjects and patients with perennial allergic rhinitis. J Laryngol otology, 1991, 105:1006～1008
28 Gary G, Geoffrey B. A novel cell-to-cell interaction between mast cells and other cell types. Exp Cell Res, 1983, 147:1～3
29 Thorp MD. The interaction between mast cells and endothelial cells, J Invest Dermatol, 1989, 93(2): 1073～1075
30 Marks RM, Roche WR. Mast cell granules cause proliferation of human mocrovascilar endothelial cells. Lab Invest, 1986, 55, 289～290
31 Cochard P, Douarin LM. Development of Instrinsic Inhervation of the Gut: Structure of the Gut. London, Glaxo Group Research Limited, 1982, 179:192～193
32 Kamagata S, Donahoe PK. The effect of fibronection on cholinergic differentiation of the fetal colon. J Pediatr Sung, 1985, 20(13): 307～308
33 Chayville JA, Paulin C, Descos F, et al. Ontogeny of vasoactive intestinal peptide in the human fetal digestive tract. Regul peptides, 1983, 5(2): 245～247
34 Paulin C, Chernay Y, Chayville JA, et al. Ontogeny of substance P in the digestive tract, spinal lord and hypothalamus of the human fetus. Regul Peptides, 1986, 14(1): 145～149
35 杨恬，蔡文琴．人胚胎期小肠六种肽能神经发生的研究．解剖学报，1994，125(1)：84～87
36 路长林．P物质．见：路长林主编，神经肽基础与临床．上海：第二军医大学出版社，2000，(1)：138～141
37 王雪琦．降钙素基因相关肽．见：路长林主编，神经肽基础与临床．上海：第二军医大学出版社，2000，(1)：195～196
38 Larsson LT. Ontogeny of peptide-containing neurons in human gut-An immunocytochemical study. Regul peptides, 1987, 17(4): 243～245
39 吴康，蔡文琴．小鼠小肠降钙素基因相关肽分布、来源和性质的研究．解剖学报，1989，20(增刊)：54～55
40 朱良如．肥大细胞与胃肠感觉和运动．国外医学消化系统疾病分册．2000，20(4)：207～208
41 Aldenbong F, Enerback L. The immunohistochemical demonstration of chymase and tryptase in human intestinal mast cells. Histochem J. 1994, 26:587～588
42 王俊明、张保真．P物质和血管活性肠肽免疫反应在人皮肤内经络线上肥大细胞颗粒内的定位研究．解剖学杂志，1986，9(增刊)：372～374
43 丰艳，吴景兰，王一菱．大鼠肥大细胞异质性免疫组织化学观察。解剖学报，1989，20(增刊)：90～94
44 贾雪梅、贾友苏、齐威琴，等．小鼠下颌下腺肥大细胞异质性的研究．1996，19(4)，344～347
45 翟力平，安巍，杨美林，等．胎儿消化器官发育中的MC超微结构特点．解剖学杂志，1997，20(1)：57～60
46 马辑研．神经肽和内啡肽对肥大细胞组胺释放的调节．国外医学免疫学分册，1992，4：171～173
2 Irani A, Schechter M, Craig S, et al. Two types of human mast cells that have distinct neutral protease composition. Proc Natl Acad Sci USA, 1986, 83: 4464～4468
3 董增军．肥大细胞亚群．国外医学免疫学分册，1990，13：185-186
4 Kitamura Y. Development of mast cell from grafted bone marrow cells in irradiated mice. Nature, 1977, 268:442～446
5 Sonda BT. Prolifetation of peritoneal mast cells in the skin of W/W~v mice that genetically lack mast cells. J Exp. Med. 1984, 160:138～140
6 Steven RL, Rothemberg ME, Levi-Schaffer F, et al. Ontogeny of in vitro differentiated mouse mast cells. Fed Proc, 1987, 46:1915～1918
7 Sonoda S, Sonoda T, Nakano T, et al. Development of mucosa mast cells after injection of a single connective tissue-type mast cell in the stomach mucosa of genetically mast cell deficient W/W~v mice. J Immunol, 1986, 137(4): 1319～1322
8 杨美林，高崇，乔从进，等，肥大细胞与成纤维细胞功能关系的光镜、电镜研究．解剖学杂志，1992，15(3)：289～291
9 Marks RM, Roche WR, Czerniecki M, et al. Mast cell granules cause proliferation of human microvascular endothelial cells. Lab Invest, 1986, 55:289～291
10 Von Euler US, Gaddum JH. An unidentified depressor substance in certain tissue extracts. J Physiol. 1931, 72: 74～78
11 邹仲之，陈东，尹昕．消化管．见：成令忠主编．组织学(第二版)．北京，人民卫生出版社．1993，1126～1139
12 朱文玉主编：神经和胃肠道肽类的关系．生理学进展．1980，11：259～261
13 李在琉．胃肠运动的神经控制．见：周吕主编．胃肠生理学——基础与临床．北京，科学出版社．1998，3(2)，363～379
14 Stead RH. Mast cells are closely apposed to nerves in the human gastrointestinal mucosa. Gastroenterology, 1989, 97:575～578
15 王俊明，张得真．P物质和血管活性肠肽免疫反应在人皮肤内经络线上肥大细胞颗粒内的定位研究．解剖学杂志．1986，9(增刊)：372～376
16 贾雪梅、贾友苏，齐威琴，等．P物质、血管活性肠肽和神经肽Y在大鼠舌肥大细胞的定位研究，解剖学杂志，1998，21(3)：242～246
17 Blennerhassett MG. Mast cells are closely apposed to nerves in the mice gastrointestinal mucosa. Cell Tissue Res, 1991, 265(1): 121～128
18 周金黄．神经免疫调节的进展．生理科学进展．1987，18(3)：119～122
19 Wingren U. Mucosal mast cells of the rat intestine: a rervaluation of fixation and staining properties with special reference to protein blocking and solubility of the granular glycosaminoglycan. Histochemical, 1983, 15: 571～572
20 何素云．消化和呼吸系统的发生．见：刘斌，高英茂主编．人体胚胎学(第二版)．北京：人民卫生出版社，1996：253～255
21 Lev R, Siegel HI, Bantman J. Histochemical studies of developing human fetal small intestine. Histochemistry, 1972, 29(2): 103～119
22 Combs JW. Differentiation and proliferation of embryonic mast cells of the rat. J cell Biol. 1965, 25:577～580
23 翟力平，杨美林．发育中人胎儿肥大细胞的组织化学研究．中国组织化学与细胞化学杂志，1995，4(2)，139～142
24 周开渠，彭庆廉．固有结缔组织．见：成令忠主编．组织学，第二版，北京：人民卫生出版社，1993，234～243
25 Craig S, Schechter M, Schwants B, Ultrastructural analysis of maturing human T and TC mast cells identified by immunoelectron. Lab Invest, 1988, 58: 682～688
26 翟力平，安巍，杨美林，等．胎儿消化器官发育中的肥大细胞超微结构特点．解剖学杂志，1997，20(1)，57～60
27 Dralce-Lee AB, Price J. Ultrastructure of mast cells in normal subjects and patients with perennial allergic rhinitis. J Laryngol otology, 1991, 105:1006～1008
28 Gary G, Geoffrey B. A novel cell-to-cell interaction between mast cells and other cell types. Exp Cell Res, 1983, 147:1～3
29 Thorp MD. The interaction between mast cells and endothelial cells, J Invest Dermatol, 1989, 93(2): 1073～1075
30 Marks RM, Roche WR. Mast cell granules cause proliferation of human mocrovascilar endothelial cells. Lab Invest, 1986, 55, 289～290
31 Cochard P, Douarin LM. Development of Instrinsic Inhervation of the Gut: Structure of the Gut. London, Glaxo Group Research Limited, 1982, 179:192～193
32 Kamagata S, Donahoe PK. The effect of fibronection on cholinergic differentiation of the fetal colon. J Pediatr Sung, 1985, 20(13): 307～308
33 Chayville JA, Paulin C, Descos F, et al. Ontogeny of vasoactive intestinal peptide in the human fetal digestive tract. Regul peptides, 1983, 5(2): 245～247
34 Paulin C, Chernay Y, Chayville JA, et al. Ontogeny of substance P in the digestive tract, spinal lord and hypothalamus of the human fetus. Regul Peptides, 1986, 14(1): 145～149
35 杨恬，蔡文琴．人胚胎期小肠六种肽能神经发生的研究．解剖学报，1994，125(1)：84～87
36 路长林．P物质．见：路长林主编，神经肽基础与临床．上海：第二军医大学出版社，2000，(1)：138～141
37 王雪琦．降钙素基因相关肽．见：路长林主编，神经肽基础与临床．上海：第二军医大学出版社，2000，(1)：195～196
38 Larsson LT. Ontogeny of peptide-containing neurons in human gut-An immunocytochemical study. Regul peptides, 1987, 17(4): 243～245
39 吴康，蔡文琴．小鼠小肠降钙素基因相关肽分布、来源和性质的研究．解剖学报，1989，20(增刊)：54～55
40 朱良如．肥大细胞与胃肠感觉和运动．国外医学消化系统疾病分册．2000，20(4)：207～208
41 Aldenbong F, Enerback L. The immunohistochemical demonstration of chymase and tryptase in human intestinal mast cells. Histochem J. 1994, 26:587～588
42 王俊明、张保真．P物质和血管活性肠肽免疫反应在人皮肤内经络线上肥大细胞颗粒内的定位研究．解剖学杂志，1986，9(增刊)：372～374
43 丰艳，吴景兰，王一菱．大鼠肥大细胞异质性免疫组织化学观察。解剖学报，1989，20(增刊)：90～94
44 贾雪梅、贾友苏、齐威琴，等．小鼠下颌下腺肥大细胞异质性的研究．1996，19(4)，344～347
45 翟力平，安巍，杨美林，等．胎儿消化器官发育中的MC超微结构特点．解剖学杂志，1997，20(1)：57～60
46 马辑研．神经肽和内啡肽对肥大细胞组胺释放的调节．国外医学免疫学分册，1992，4：171～173