慢传输型便秘大鼠血浆及肠道胃动素变化的实验研究
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摘要
目的:本研究课题利用大黄灌胃建立慢传输型便秘大鼠模型,通过测定模型大鼠血浆及十二指肠、回肠、横结肠组织匀浆中胃动素(MTL)含量,观察模型大鼠肠道胃动素免疫反应性变化,探讨血浆及肠道MTL水平与慢传输型便秘的关系。
方法:1、动物分组:选取健康Wistar大鼠40只,雌雄各半,按照完全随机的方法进行分组,实验组20只,对照组20只,分组后各组间体重、性别无显著性差异。
2、动物处理:实验组采用大黄浸液灌胃,首日剂量为800mg/kg(生粉) ,此后每日剂量在前一日基础上增加200mg/kg(生粉),直至实验组出现半数大鼠粪便变稀,保持剂量至80%的稀便消失,继续加量给药,至又有半数大鼠粪便变稀,如此程序循环3次,待实验组第三次80%的稀便消失一周后停止建模,对照组以同等体积蒸馏水灌胃。
3、标本取材:实验组及对照组大鼠禁食8小时后乙醚麻醉,股动脉取抗凝血3ml,切取距幽门5cm十二指肠、距回盲瓣5cm回肠、横结肠中段组织各两块,每块长约1cm,其中一块组织多聚甲醛固定,包埋切片后进行HE染色、MTL免疫组织化学染色,观察免疫反应阳性细胞数量及形态变化。另一块组织称重,匀浆,应用荧光分光光度法测量血浆及组织匀浆中MTL含量。
4、半定量分析:根据MTL免疫反应阳性细胞的分布密度及活动性强弱,做相对半定量比较。所有视野未见阳性细胞为(-);有少数阳性细胞为(+),染色为淡黄色;阳性细胞较多见为(++),染色为棕黄色;阳性细胞多见为(+++),染色为棕褐色。
5、图像分析:在CMIAS系列多功能病理图像分析系统下,每组选5例染色良好的切片,在显微镜物镜放大10倍下观察,每张切片随机选取5个视野,共25个视野摄入计算机(分辨率为800×600),测量每个视野阳性染色区域的表面积密度,即面密度。最后计算每组的平均值。并用尼康光镜照相机拍摄典型视野。
6、统计分析:本实验计数资料数据采用X±s表示,应用SPSS10.0统计软件,选用成组t检验进行统计分析,检验水准α=0.05。半定量分析结果采用Excel编程统计,检验方法选用成组设计多个样本比较的秩和检验(Kruskal-Wallis法),检验水准α=0.05。
结果:1、肠道大体病理变化及HE染色:两组大鼠肠道外观无明显差别,结肠粘膜无色素沉着。对照组大鼠肠道切片HE染色未发现病变,实验组大鼠肠道切片HE染色可见粘膜慢性炎症改变。
2、MTL免疫组织化学结果:肠道的MTL免疫组织化学染色显示其阳性产物呈棕黄色沉淀。十二指肠、回肠粘膜及粘膜下层可见MTL免疫反应阳性细胞分布, MTL免疫反应阳性细胞呈圆形,着色淡。实验组及对照组十二指肠肌层及肌间神经丛、回肠肌层及肌间神经丛以及横结肠全层未见阳性细胞及阳性纤维。实验组十二指肠MTL免疫反应性较对照组明显降低(p<0.05);实验组回肠MTL免疫反应性较对照组明显降低(P<0.05)。
对MTL免疫组织化学图像分析的结果与半定量分析一致。
3、血浆中MTL含量:实验组大鼠血浆中MTL含量较对照组降低(p<0.05)。
4、肠道组织匀浆中MTL含量:实验组及对照组大鼠十二指肠组织匀浆中MTL含量均高于同组回肠组织匀浆中MTL含量(p<0.05);实验组大鼠十二指肠组织匀浆中MTL含量较对照组十二指肠组织匀浆中MTL含量降低(p<0.05),实验组大鼠回肠组织匀浆中MTL含量较对照组回肠组织匀浆中MTL含量降低(p<0.05);实验组及对照组大鼠横结肠组织匀浆中MTL均未测出。
结论:1、应用大黄灌胃制造STC大鼠模型更为精确,是探讨STC发病机制的一个较好模型。
2、大鼠MTL主要集中在近端小肠,在结肠中没有分布。慢传输型便秘大鼠血浆与肠道粘膜MTL含量及免疫反应性降低,血浆及肠道粘膜MTL水平与STC有一定相关性。
Objective: This experiment established a rat’s slow transit constipation model by rhubarb intragastric administration. When model constructed, we detected the content of motilin (MTL) in homogenate of duodenum, ileum and transverse colon and in blood plasma, observed the change of immunological reactivity of MTL in model rats’intestines, and investigated the relation-ship between slow transit constipation and the level of MTL in blood plasma and intestine.
Method:40 clean healthy adult Wister rats, half female and half male respectively, were chosen and divided randomly into control group and rhubarb group, 20 each group. Marked weight and sex difference did not exist after grouping.
In rhubarb groups, leachate of rhubarb was administered intragastricly. At the beginning, the dosage was 800mg/kg/day (raw material), afterwards the dosage increased 200mg/kg/day (raw material) every day until half rats’stool became watery, this dosage was maintained until the watery stool of 80 percent rats disappeared, then the dosage continually increased until 50 percent of rats’stool become watery. One week after the watery stool of 80 percent rats disappeared for the third time, the ex-periment was stopped. Distilled water was administered intra- gastricly in control group.
After model constructed, we anesthetized all rats with di-ethyl ether and drew 3ml anticoagulated blood from femoral artery. After the abdominal cavity opened quickly, we cut two 1cm blocks of tissues from duodenum 5cm to pylorus, ileum 5cm to ileocecal valve and midpiece of transverse colon, respec-tively. One tissue block was fixed with Polyoxymethylene, then was stained by means of HE stain and MTL immunohisto-chemical stain after embedded and cut to sections for observing the morphologic change and number of the positive cells. Be-hind the other block weighted and homogenized, we detected the content of MTL in homogenate and in blood plasma by spectrofluorimetry.
According to the distribution and activity of the cell, in which MTL expressed positively, the results were compared hemi-quantitatively. Staining extent was scored as“–”(none),“+”(few, amber),“++”(many, buffy),“+++”(lot, brown) ac-cording to the numbers of the positive cells in entire section.
Five well-stained sections, which were selected from each group, were observed under a light microscope at low magnifi-cation (10×), and five visual fields were selected randomly in each section. Then the twenty-five visual fields were input to computer (800×600) and density of surface area of posi-tive-stained region was measured in each section. At last, we calculated the average of each group and photographed typical visual field with Nikon optical camera.
This experiment’s measure data was expressed by X±s, adopting SPSS 10.0 statistics software, chose two-group t-test, test levelα=0.05. The result of hemi-quantitative analysis was analyzed statistically with Excle, chose Kruskal-Wallis H test, test levelα=0.05.
Result: In the rhubarb group, the appearance of gastroin-testinal tract altered lightly and the colonic mucosa had no pig-mentation. HE staining had no chronic inflammation in control group while reflected in rhubarb group.
MTL immunohistochemical stain displayed that positive products shew buffy. In the submucosa of duodenum and ileum, we could see MTL+ cells and positive fibers, and the MTL+ cells in mucous membrane were round and light colored. MTL hardly expressed in transverse colon in muscular layer and myenteric nerve plexus of duodenum and ileum in both of control group and rhubarb group.
Rhubarb group had much lower immunoreactivity of MTL in duodenum than control group (P<0.05); and the same result was shown in ileum (P<0.05). There had no difference between the result of immunohistochemical image analysis and hemi-quantitative analysis of MTL.
In rhubarb group, the content of MTL in blood plasma was lower than those in control group (P<0.05). In both of control group and rhubarb group, the content of MTL in duodenum homogenate was higher than in ileum homogenate (P<0.05), and rhubarb group had fewer MTL in homogenate of duodenum and ileum than control group (P<0.05). No MTL was detected in homogenate of transverse colon in both groups.
Conclusion: The slow transit constipation model induced by the long-term intragastric administration of rhubarb is ex-acter than by others means, and is a better model for investigat-ing the pathogenesis of slow transit constipation.
For normal rats, MTL expresses in proximal small bowel predominantly, and does not express in colon. The contents of MTL is low in blood plasma and mucous membrane of intestinal tract of rats, it shows that the level of MTL in blood plasma and mucous membrane of intestinal tract has some association with slow transit constipation.
方法:1、动物分组:选取健康Wistar大鼠40只,雌雄各半,按照完全随机的方法进行分组,实验组20只,对照组20只,分组后各组间体重、性别无显著性差异。
2、动物处理:实验组采用大黄浸液灌胃,首日剂量为800mg/kg(生粉) ,此后每日剂量在前一日基础上增加200mg/kg(生粉),直至实验组出现半数大鼠粪便变稀,保持剂量至80%的稀便消失,继续加量给药,至又有半数大鼠粪便变稀,如此程序循环3次,待实验组第三次80%的稀便消失一周后停止建模,对照组以同等体积蒸馏水灌胃。
3、标本取材:实验组及对照组大鼠禁食8小时后乙醚麻醉,股动脉取抗凝血3ml,切取距幽门5cm十二指肠、距回盲瓣5cm回肠、横结肠中段组织各两块,每块长约1cm,其中一块组织多聚甲醛固定,包埋切片后进行HE染色、MTL免疫组织化学染色,观察免疫反应阳性细胞数量及形态变化。另一块组织称重,匀浆,应用荧光分光光度法测量血浆及组织匀浆中MTL含量。
4、半定量分析:根据MTL免疫反应阳性细胞的分布密度及活动性强弱,做相对半定量比较。所有视野未见阳性细胞为(-);有少数阳性细胞为(+),染色为淡黄色;阳性细胞较多见为(++),染色为棕黄色;阳性细胞多见为(+++),染色为棕褐色。
5、图像分析:在CMIAS系列多功能病理图像分析系统下,每组选5例染色良好的切片,在显微镜物镜放大10倍下观察,每张切片随机选取5个视野,共25个视野摄入计算机(分辨率为800×600),测量每个视野阳性染色区域的表面积密度,即面密度。最后计算每组的平均值。并用尼康光镜照相机拍摄典型视野。
6、统计分析:本实验计数资料数据采用X±s表示,应用SPSS10.0统计软件,选用成组t检验进行统计分析,检验水准α=0.05。半定量分析结果采用Excel编程统计,检验方法选用成组设计多个样本比较的秩和检验(Kruskal-Wallis法),检验水准α=0.05。
结果:1、肠道大体病理变化及HE染色:两组大鼠肠道外观无明显差别,结肠粘膜无色素沉着。对照组大鼠肠道切片HE染色未发现病变,实验组大鼠肠道切片HE染色可见粘膜慢性炎症改变。
2、MTL免疫组织化学结果:肠道的MTL免疫组织化学染色显示其阳性产物呈棕黄色沉淀。十二指肠、回肠粘膜及粘膜下层可见MTL免疫反应阳性细胞分布, MTL免疫反应阳性细胞呈圆形,着色淡。实验组及对照组十二指肠肌层及肌间神经丛、回肠肌层及肌间神经丛以及横结肠全层未见阳性细胞及阳性纤维。实验组十二指肠MTL免疫反应性较对照组明显降低(p<0.05);实验组回肠MTL免疫反应性较对照组明显降低(P<0.05)。
对MTL免疫组织化学图像分析的结果与半定量分析一致。
3、血浆中MTL含量:实验组大鼠血浆中MTL含量较对照组降低(p<0.05)。
4、肠道组织匀浆中MTL含量:实验组及对照组大鼠十二指肠组织匀浆中MTL含量均高于同组回肠组织匀浆中MTL含量(p<0.05);实验组大鼠十二指肠组织匀浆中MTL含量较对照组十二指肠组织匀浆中MTL含量降低(p<0.05),实验组大鼠回肠组织匀浆中MTL含量较对照组回肠组织匀浆中MTL含量降低(p<0.05);实验组及对照组大鼠横结肠组织匀浆中MTL均未测出。
结论:1、应用大黄灌胃制造STC大鼠模型更为精确,是探讨STC发病机制的一个较好模型。
2、大鼠MTL主要集中在近端小肠,在结肠中没有分布。慢传输型便秘大鼠血浆与肠道粘膜MTL含量及免疫反应性降低,血浆及肠道粘膜MTL水平与STC有一定相关性。
Objective: This experiment established a rat’s slow transit constipation model by rhubarb intragastric administration. When model constructed, we detected the content of motilin (MTL) in homogenate of duodenum, ileum and transverse colon and in blood plasma, observed the change of immunological reactivity of MTL in model rats’intestines, and investigated the relation-ship between slow transit constipation and the level of MTL in blood plasma and intestine.
Method:40 clean healthy adult Wister rats, half female and half male respectively, were chosen and divided randomly into control group and rhubarb group, 20 each group. Marked weight and sex difference did not exist after grouping.
In rhubarb groups, leachate of rhubarb was administered intragastricly. At the beginning, the dosage was 800mg/kg/day (raw material), afterwards the dosage increased 200mg/kg/day (raw material) every day until half rats’stool became watery, this dosage was maintained until the watery stool of 80 percent rats disappeared, then the dosage continually increased until 50 percent of rats’stool become watery. One week after the watery stool of 80 percent rats disappeared for the third time, the ex-periment was stopped. Distilled water was administered intra- gastricly in control group.
After model constructed, we anesthetized all rats with di-ethyl ether and drew 3ml anticoagulated blood from femoral artery. After the abdominal cavity opened quickly, we cut two 1cm blocks of tissues from duodenum 5cm to pylorus, ileum 5cm to ileocecal valve and midpiece of transverse colon, respec-tively. One tissue block was fixed with Polyoxymethylene, then was stained by means of HE stain and MTL immunohisto-chemical stain after embedded and cut to sections for observing the morphologic change and number of the positive cells. Be-hind the other block weighted and homogenized, we detected the content of MTL in homogenate and in blood plasma by spectrofluorimetry.
According to the distribution and activity of the cell, in which MTL expressed positively, the results were compared hemi-quantitatively. Staining extent was scored as“–”(none),“+”(few, amber),“++”(many, buffy),“+++”(lot, brown) ac-cording to the numbers of the positive cells in entire section.
Five well-stained sections, which were selected from each group, were observed under a light microscope at low magnifi-cation (10×), and five visual fields were selected randomly in each section. Then the twenty-five visual fields were input to computer (800×600) and density of surface area of posi-tive-stained region was measured in each section. At last, we calculated the average of each group and photographed typical visual field with Nikon optical camera.
This experiment’s measure data was expressed by X±s, adopting SPSS 10.0 statistics software, chose two-group t-test, test levelα=0.05. The result of hemi-quantitative analysis was analyzed statistically with Excle, chose Kruskal-Wallis H test, test levelα=0.05.
Result: In the rhubarb group, the appearance of gastroin-testinal tract altered lightly and the colonic mucosa had no pig-mentation. HE staining had no chronic inflammation in control group while reflected in rhubarb group.
MTL immunohistochemical stain displayed that positive products shew buffy. In the submucosa of duodenum and ileum, we could see MTL+ cells and positive fibers, and the MTL+ cells in mucous membrane were round and light colored. MTL hardly expressed in transverse colon in muscular layer and myenteric nerve plexus of duodenum and ileum in both of control group and rhubarb group.
Rhubarb group had much lower immunoreactivity of MTL in duodenum than control group (P<0.05); and the same result was shown in ileum (P<0.05). There had no difference between the result of immunohistochemical image analysis and hemi-quantitative analysis of MTL.
In rhubarb group, the content of MTL in blood plasma was lower than those in control group (P<0.05). In both of control group and rhubarb group, the content of MTL in duodenum homogenate was higher than in ileum homogenate (P<0.05), and rhubarb group had fewer MTL in homogenate of duodenum and ileum than control group (P<0.05). No MTL was detected in homogenate of transverse colon in both groups.
Conclusion: The slow transit constipation model induced by the long-term intragastric administration of rhubarb is ex-acter than by others means, and is a better model for investigat-ing the pathogenesis of slow transit constipation.
For normal rats, MTL expresses in proximal small bowel predominantly, and does not express in colon. The contents of MTL is low in blood plasma and mucous membrane of intestinal tract of rats, it shows that the level of MTL in blood plasma and mucous membrane of intestinal tract has some association with slow transit constipation.
引文
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30 黄裕新,许才绂,张少玲等.血浆胃动素与小肠传递时间关系的探讨[J].陕西医学杂志,1993,22(1):9～10
31 罗金燕,郭剑峰,董雪等.肠易激综合征患者的胃肠时间测定[J].中华消化杂志,1994,14 (增刊):39～41
1 Ddrossman DA, Corazziari E, Jalley NJ , et al. Functional gastrointinal disorders: Rome Ⅱ [M]. Mclean Virginia: Dgnon and Associates, 2000
2 Archar P,Deveroede G, Jehannin G, et al. Segmental colonic transit time[J]. Dis Colon Rectum, 1981, 24: 625
3 P.Facer. Decreased tyrosine kinase C expression may re-flect derelopmental abnormalities in Hirsch sprung’s Disease and idiopathic slow transit constipation. Br J Surg, 2001,88: 545～552
4 D.Mitolo-chieppa.Idiopathic chronic constipation: tachy- kinins as cotransmitters in colonic contraction. European Journal of Clinical Inrestigation , 2001,31:349～355
5 Brown JC, etal. Effect of duodenal alkalinization on gastric motility Gastroenterology, 1966,50:333～339
6 周吕.胃肠生理学[M].北京:北京科学出版,1991.624～626.
7 黄裕新,许才绂,冯英明. 胃动素在人体13 种体液中分布及关系的研究[J].中华消化杂志,1994 ,14 (2):96
8 谷成明,宋巧云.胃动素 北京军区医药 1994,6(6)459～461
9 Palak JM. Complete identification of endocrine cells in the gastrointestinal tract using se mithin sections to identify mo-tilin cells in human and animal intestine. [J].Cut, 1995, 16(2): 225～230
10 陈泮藻,李振甲,郝秀华等.胃动素快速放射免疫分析及初步临床应用[J].中华医学检验杂志,1990, 13(5):279～281
11 黄裕新,许才绂,冯英明等.西安市正常人空腹血浆胃动素含量放免测定[J].第四军医大学学报,1991,12(6): 477～478
12 Jenssen TG,Burhol PG,Jorde R,etal.RIA ;pals ma motilin in man [J].ScardJ Gastroenterrol, 1984,19 (6):717～723
13 Christofieds ND,Bloom SR,Besterman S,Physiologyof mo-tilinl l [M].In:Bloom SR,ED.Gut hormones [M].2 nded.Edinburgh:Churchill Livingsone,1978.343～350
14 王志均.胃肠激素[M].北京:科学出版社,1985.210～307
15 周吕,柯美云.胃肠动力学基础与临床[M].北京:科学出版社,1994.126～149
16 徐胜生,焦俊,曾曦,张兰花.结肠运动与血胃动素、血管活性肠肽和P物质的关系 贵阳医学院学报,2004 (10) 416～420
17 姚永莉,徐波.胃肠道功能紊乱与胃动素关系的实验研究 第一军医大学学报 2001,21(8)602～603
18 Sjolund K,Ekman R, Akre F, et al. Motilin in chronic dio-pathic constipation [J].Scand J Gastroenterd,1986, 21:914～918
19 Stern HP,Stroh SE,Fiedorek SC,et al.Increased plas ma lev-els of motilin in encopretic children [J].Pediatrics,1995, 96:111-117
20 Preston DM, Adrian TE, Christofides ND, et al .Positive correlation between symptoms and circulationg motilin. pancreatic polypeptides and gastrin levels in functional bowel disorders[J].Gut ,1985,26:1059-1064
21 Joost R,Van der Sijp,Michael A,et al.Circulating gastrointes-tinal hormone abnormalities in patients with severe idio-pathic constipation [J].Scand J Gastroenterology, 1998,93:1351-1356
22 Peracchi M, Basilisco G, Tagliabue R, et al . Postprandial gut peptide plasma levels inwoman with idiopathic slow transit constipation[J].Scand J Gastroenterol.1999,34(1):25～28
23 张兰花,焦俊,郭晓山.慢传输型便秘患者结肠动力与胃肠激素的关系 临床消化病杂志 2005,17(1)17～19
24 姜若兰.胃肠激素与胃肠运动[J].中华消化杂志,1996, 16(6):353～355
25 蔡昌豪,王孟薇.胃动素研究进展[J].解放军进修学院学报,1991,12(4):337～339
26 黄裕新,许才绂,张少玲等.血浆胃动素与小肠传递时间关系的探讨[J].陕西医学杂志,1993,22(1):9～10
27 罗金燕,郭剑峰,董雪等.肠易激综合征患者的胃肠时间测定[J].中华消化杂志,1994,14 (增刊):39～41
28 Kachel GW. Effect of 13 - horleucin motilin on water and serummotilin levels in patients with idiopathic delay in gas-tric emptying [J] . Gastroentrology, 1986 ,90 :20
29 柳力公,周吕,王玲.肠神经系统脑肠肽神经元的免疫组化研究.基础医学与临床,2003,23:348～349
30 周吕,王礼建.胃动素对 Cajal 间质细胞引起的大鼠胃平滑肌收缩的作用. 中华医学杂志 2003,83(16):1422～1427
31 Feighner SD,Tan CP,McKee KK,et al.Receptor for motilin identified in the human gastrointestinal system[J]. Science, 1999,284(5423): 2184～2188.
32 Van Assche G, Depoortere I, Thijs T, et al. Contractile effects and intracellular Ca2+ signalling induced by motilin and erythromycin in the circular smooth muscle of human colon. Neurogastroentero Motil, 2001, 13 (1) : 27～35
33 Matthijs G, Peeters TL , Vantrappen G. Effect of different ofcalcium modulators on motilin-induced contractions of the rabbit duodenum. Comparison with acetylcho line. Regul Pep t, 1988; 21 (324): 321～330
34 Depoortere I, Peeters TL. Transduction mechanism of mo-tilin and motilides in rabbit duodenal smooth muscle. Regul Pep t, 1995; 55 (3): 227～235
35 刘梅,董蕾,朱文艺.大鼠杏仁核胃动素受体分布及其作用研究. 第一军医大学学报, 2005; 25 (9): 1100～1105
36 Zhou X, Galligan JJ. P2X purinocep to rs in cultured myenteric neurons of guinea-pig small intestine. J Physio l, 1996; 496(p t3): 719～729
37 Depoortere I, Thijs T, Thielemans L, et al. Interaction of the growth hormone-releasing pep tides-6 with the motilin re-ceptor in the rabbit gastric antrum. J Pharmaco l Exp Ther, 2003; 305(2): 660～667
38 VanA ssche G, Depoortere I, Thijs T, et al. Concentration- dependent stimulation of cholinergic motor nerves or smooth muscle by [Nle 13] motilin in the isolated rabbit gastric antrum.Eur J Pharmaco l, 1997; 337 (223) : 267～274
39 Bisschop s R, V anden Berghe P, Depoortere I, et al. Ghrelin activates a subset of myenteric neurons in the guinea pig je-junum. Gastroentero logy, 2003;124 (1): A 1
40 Miller P, Trudel L S, Pierre ST, et al. Neural and muscular recep to rs for motilin in the rabbit colon. Pep tides, 2000; 21(2): 283～287
41 张绍刚,程留芳,郭荣斌等.习惯性便秘患者胃肠激素的变化.河北医药,2000;22(9):664～665
42 Stem HP ,Stroh SE ,Fiedorek SC ,et al .Increased plasma levels of pancreatic polypeptide and decresade plasms levels of motilin in encopretic children ,Rayford PL Pediatrics, 1995;96(1Pt 1):111～117
2 李海燕 慢传输型便秘发病机理研究进展 国外医学·消化系疾病分册 2004,24(6) 366～369
3 Porrceca F,Galligan JJ,Burks TF. Central opioid receptor involve mention gastrointestinal motility [J]. Trends Phar-macol Sci , 1986 ,7 :104
4 张连阳等.一种大鼠“泻剂结肠”模型的建立.大肠肛门病外科杂志,1998,3:66～68
5 杨士友,黄世福,田军.不同提取方法对大黄泻下成分的影响.安徽医药, 2004,2:9(2)
6 王颢,喻德洪 慢传输型便秘的发病机制研究近况 中国现代普通外科进展 2001,4(2)79～81
7 Ddrossman DA, Corazziari E, Jalley NJ, et al. Functional gastrointinal disorders: Rome Ⅱ [M]. Mclean Virginia: Dgnon and Associates, 2000
8 Archar P,Deveroede G, Jehannin G, et al. Segmental colonic transit time[ J ]. Dis Colon Rectum, 1981, 24: 625
9 P.Facer. Decreased tyrosine kinase C expression may re-flect derelopmental abnormalities in Hirsch sprung’s Disease and idiopathic slow transit constipation. Br J Surg, 2001;88:545～552
10 D.Mitolo-chieppa.Idiopathic chronic constipation: tachy-kinins as cotransmitters in colonic contraction. Euro-pean Journal of Clinical Inrestigation,2001;31: 349～ 355
11 柳力公,周吕,王玲.肠神经系统脑肠肽神经元的免疫组化研究.基础医学与临床,2003,23:348～349
12 周吕,王礼建,袁勃等.胃动素对 Cajal 间质细胞引起的大鼠胃平滑肌收缩的作用中华医学杂志 2003;83(16);1422～1427
13 Brown JC, etal. Effect of duodenal alkalinization on gastric motility Gastroenterology, 1966,50: 333～339
14 周吕.胃肠生理学[M].北京:北京科学出版社,1991.624 ～626
15 黄裕新,许才绂,冯英明. 胃动素在人体 13 种体液中分布及关系的研究[J] . 中华消化杂志,1994 ,14(2):96
16 谷成明,宋巧云.胃动素 北京军区医药 1994,6(6)459～461
17 Palak JM. Complete identification of endocrine cells in the gastrointestinal tract using se mithin sections to identify mo-tilin cells in human and animal intestine. [J].Cut,1995, 16(2): 225～230
18 王志均.胃肠激素[M].北京:科学出版社,1985.210～307
19 Feighner SD,Tan CP,McKee KK,et al.Receptor for motilin identified in the human gastrointestinal system[J]. Science, 1999,284(5423): 2184～2188
20 Van Assche G, Depoortere I, Thijs T, et al. Contractile effects and intracellular Ca2+ signalling induced by motilin and erythromycin in the circular smooth muscle of human colon. Neurogastroentero Motil,2001;13(1): 27～35
21 Matthijs G, Peeters TL, Vantrappen G. Effect of different of calcium modulators on motilin-induced contractions of the rabbit duodenum. Comparison with acetylcho line. Regul Pep t, 1988; 21 (324): 321～330
22 Depoortere I, Peeters TL. Transduction mechanism of mo-tilin and motilides in rabbit duodenal smooth muscle. Regul Pep t, 1995; 55 (3) : 227～235
23 刘梅,董蕾,朱文艺.大鼠杏仁核胃动素受体分布及其作用研究. 第一军医大学学报, 2005; 25(9): 1100～1105
24 Zhou X, Galligan JJ. P2X purinocep to rs in cultured myenteric neurons of guinea-pig small intestine. J Physio l, 1996; 496(p t3): 719～729
25 Depoortere I, Thijs T, Thielemans L, et al. Interaction of the growth hormone-releasing pep tides-6 with the motilin re-ceptor in the rabbit gastric antrum. J Pharmaco l Exp Ther, 2003; 305(2): 660～667
26 VanA ssche G, Depoortere I, Thijs T, et al. Concentration dependent stimulation of cholinergic motor nerves or smooth muscle by [N le13] motilin in the isolated rabbit gastric antrum.Eur J Pharmaco l, 1997; 337 (223) : 267～274
27 Bisschop s R, V anden Berghe P, Depoortere I, et al. Ghrelin activates a subset of myenteric neurons in the guinea pig je-junum. Gastroentero logy, 2003;124 (1): A 1
28 Miller P, Trudel L S, Pierre ST, et al. Neural and muscular recep to rs for motilin in the rabbit colon. Pep tides, 2000; 21(2): 283～287
29 蔡昌豪,王孟薇.胃动素研究进展[J].解放军进修学院学报,1991,12(4):337～339
30 黄裕新,许才绂,张少玲等.血浆胃动素与小肠传递时间关系的探讨[J].陕西医学杂志,1993,22(1):9～10
31 罗金燕,郭剑峰,董雪等.肠易激综合征患者的胃肠时间测定[J].中华消化杂志,1994,14 (增刊):39～41
1 Ddrossman DA, Corazziari E, Jalley NJ , et al. Functional gastrointinal disorders: Rome Ⅱ [M]. Mclean Virginia: Dgnon and Associates, 2000
2 Archar P,Deveroede G, Jehannin G, et al. Segmental colonic transit time[J]. Dis Colon Rectum, 1981, 24: 625
3 P.Facer. Decreased tyrosine kinase C expression may re-flect derelopmental abnormalities in Hirsch sprung’s Disease and idiopathic slow transit constipation. Br J Surg, 2001,88: 545～552
4 D.Mitolo-chieppa.Idiopathic chronic constipation: tachy- kinins as cotransmitters in colonic contraction. European Journal of Clinical Inrestigation , 2001,31:349～355
5 Brown JC, etal. Effect of duodenal alkalinization on gastric motility Gastroenterology, 1966,50:333～339
6 周吕.胃肠生理学[M].北京:北京科学出版,1991.624～626.
7 黄裕新,许才绂,冯英明. 胃动素在人体13 种体液中分布及关系的研究[J].中华消化杂志,1994 ,14 (2):96
8 谷成明,宋巧云.胃动素 北京军区医药 1994,6(6)459～461
9 Palak JM. Complete identification of endocrine cells in the gastrointestinal tract using se mithin sections to identify mo-tilin cells in human and animal intestine. [J].Cut, 1995, 16(2): 225～230
10 陈泮藻,李振甲,郝秀华等.胃动素快速放射免疫分析及初步临床应用[J].中华医学检验杂志,1990, 13(5):279～281
11 黄裕新,许才绂,冯英明等.西安市正常人空腹血浆胃动素含量放免测定[J].第四军医大学学报,1991,12(6): 477～478
12 Jenssen TG,Burhol PG,Jorde R,etal.RIA ;pals ma motilin in man [J].ScardJ Gastroenterrol, 1984,19 (6):717～723
13 Christofieds ND,Bloom SR,Besterman S,Physiologyof mo-tilinl l [M].In:Bloom SR,ED.Gut hormones [M].2 nded.Edinburgh:Churchill Livingsone,1978.343～350
14 王志均.胃肠激素[M].北京:科学出版社,1985.210～307
15 周吕,柯美云.胃肠动力学基础与临床[M].北京:科学出版社,1994.126～149
16 徐胜生,焦俊,曾曦,张兰花.结肠运动与血胃动素、血管活性肠肽和P物质的关系 贵阳医学院学报,2004 (10) 416～420
17 姚永莉,徐波.胃肠道功能紊乱与胃动素关系的实验研究 第一军医大学学报 2001,21(8)602～603
18 Sjolund K,Ekman R, Akre F, et al. Motilin in chronic dio-pathic constipation [J].Scand J Gastroenterd,1986, 21:914～918
19 Stern HP,Stroh SE,Fiedorek SC,et al.Increased plas ma lev-els of motilin in encopretic children [J].Pediatrics,1995, 96:111-117
20 Preston DM, Adrian TE, Christofides ND, et al .Positive correlation between symptoms and circulationg motilin. pancreatic polypeptides and gastrin levels in functional bowel disorders[J].Gut ,1985,26:1059-1064
21 Joost R,Van der Sijp,Michael A,et al.Circulating gastrointes-tinal hormone abnormalities in patients with severe idio-pathic constipation [J].Scand J Gastroenterology, 1998,93:1351-1356
22 Peracchi M, Basilisco G, Tagliabue R, et al . Postprandial gut peptide plasma levels inwoman with idiopathic slow transit constipation[J].Scand J Gastroenterol.1999,34(1):25～28
23 张兰花,焦俊,郭晓山.慢传输型便秘患者结肠动力与胃肠激素的关系 临床消化病杂志 2005,17(1)17～19
24 姜若兰.胃肠激素与胃肠运动[J].中华消化杂志,1996, 16(6):353～355
25 蔡昌豪,王孟薇.胃动素研究进展[J].解放军进修学院学报,1991,12(4):337～339
26 黄裕新,许才绂,张少玲等.血浆胃动素与小肠传递时间关系的探讨[J].陕西医学杂志,1993,22(1):9～10
27 罗金燕,郭剑峰,董雪等.肠易激综合征患者的胃肠时间测定[J].中华消化杂志,1994,14 (增刊):39～41
28 Kachel GW. Effect of 13 - horleucin motilin on water and serummotilin levels in patients with idiopathic delay in gas-tric emptying [J] . Gastroentrology, 1986 ,90 :20
29 柳力公,周吕,王玲.肠神经系统脑肠肽神经元的免疫组化研究.基础医学与临床,2003,23:348～349
30 周吕,王礼建.胃动素对 Cajal 间质细胞引起的大鼠胃平滑肌收缩的作用. 中华医学杂志 2003,83(16):1422～1427
31 Feighner SD,Tan CP,McKee KK,et al.Receptor for motilin identified in the human gastrointestinal system[J]. Science, 1999,284(5423): 2184～2188.
32 Van Assche G, Depoortere I, Thijs T, et al. Contractile effects and intracellular Ca2+ signalling induced by motilin and erythromycin in the circular smooth muscle of human colon. Neurogastroentero Motil, 2001, 13 (1) : 27～35
33 Matthijs G, Peeters TL , Vantrappen G. Effect of different ofcalcium modulators on motilin-induced contractions of the rabbit duodenum. Comparison with acetylcho line. Regul Pep t, 1988; 21 (324): 321～330
34 Depoortere I, Peeters TL. Transduction mechanism of mo-tilin and motilides in rabbit duodenal smooth muscle. Regul Pep t, 1995; 55 (3): 227～235
35 刘梅,董蕾,朱文艺.大鼠杏仁核胃动素受体分布及其作用研究. 第一军医大学学报, 2005; 25 (9): 1100～1105
36 Zhou X, Galligan JJ. P2X purinocep to rs in cultured myenteric neurons of guinea-pig small intestine. J Physio l, 1996; 496(p t3): 719～729
37 Depoortere I, Thijs T, Thielemans L, et al. Interaction of the growth hormone-releasing pep tides-6 with the motilin re-ceptor in the rabbit gastric antrum. J Pharmaco l Exp Ther, 2003; 305(2): 660～667
38 VanA ssche G, Depoortere I, Thijs T, et al. Concentration- dependent stimulation of cholinergic motor nerves or smooth muscle by [Nle 13] motilin in the isolated rabbit gastric antrum.Eur J Pharmaco l, 1997; 337 (223) : 267～274
39 Bisschop s R, V anden Berghe P, Depoortere I, et al. Ghrelin activates a subset of myenteric neurons in the guinea pig je-junum. Gastroentero logy, 2003;124 (1): A 1
40 Miller P, Trudel L S, Pierre ST, et al. Neural and muscular recep to rs for motilin in the rabbit colon. Pep tides, 2000; 21(2): 283～287
41 张绍刚,程留芳,郭荣斌等.习惯性便秘患者胃肠激素的变化.河北医药,2000;22(9):664～665
42 Stem HP ,Stroh SE ,Fiedorek SC ,et al .Increased plasma levels of pancreatic polypeptide and decresade plasms levels of motilin in encopretic children ,Rayford PL Pediatrics, 1995;96(1Pt 1):111～117