旋复代赭汤对胃动力低下大鼠胃动力影响的实验研究
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摘要
目的:胃肠动力障碍是由多种原因所导致的以胃排空延迟和小肠推进减慢为特征的一组病证,它是众多功能性消化道疾病的共同病理变化。此外,有些器质性消化道疾病如消化性溃疡、慢性胃炎及胃癌等,可同时伴有胃肠动力障碍。现代社会,因竞争加剧,生活节奏加快,人们的精神愈加紧张,常可导致迷走神经的兴奋性增强,使胃肠动力障碍性疾病的发生呈上升趋势。近年来,国外学者围绕着消化道动力障碍的防治,广泛开展了促胃肠动力障碍药物的研究。在发达国家,每年平均用于研究开发促胃肠动力新药的耗资达10亿美元,已有胃复安、吗丁啉、西沙比利及莫沙比利等药物应用于临床。但临床使用表明,这些促胃肠动力西药的疗效和应用范围尚欠理想,并且还存在中枢神经系统、循环系统及其它副作用。目前我国也没有开发出理想的促胃肠动力药。临床研究发现,旋复代赭汤对各种胃肠动力障碍疾病的治疗有显著的疗效。本课题用一定剂量的甘草煎剂制作胃动力低下大鼠模型,观察旋复代赭汤对胃动力低下大鼠血液及胃窦组织中脑肠肽含量的影响、胃窦组织中5-HT APUD细胞数目及脑肠肽受体mRNA表达情况等的影响,为拓宽本方的临床应用和药物开发提供科学的理论依据。
方法:先用甘草煎剂制作胃动力低下模型:将Wistar大鼠30只,随机分为正常组、模型3天组及模型7天组,每组10只,雌雄各半。用一定剂量的甘草煎剂灌胃给药后,用放射性核素标记法检测大鼠的胃半排时间。再将健康Wistar大鼠70只随机分为正常组和模型组,正常组10只,模型组60只,雌雄各半。模型组每只大鼠按10g/kg体重的甘草煎剂灌胃,第三次给药前24小时动物禁食不禁水。第三天给药后,将60只大鼠随机分为模型3天组、模型7天组、旋复代赭汤低、中、高剂量组、吗丁啉组。每组10只,雌雄各半。除了模型3天组外,从第四天开始,模型7天组、旋复代赭汤各剂量组及吗丁啉组继续用甘草煎剂予上午8:00灌胃,直至第7天结束;从第三天开始,下午4:00正常组及模型7天组予等体积的生理盐水灌胃,吗丁啉组予0.27mg/ml的吗丁啉混悬液按100g体重1ml灌胃,中药低、中、高各剂量组分别予浓度为0.369g/ml、0.738g/ml、1.476g/ml的旋复代赭汤灌胃,共给药5天,末次给药前24小时动物禁食不禁水。用放免法观测各组大鼠血液及组织中脑肠肽的含量;用免疫组化法检测胃窦组织中5-HT APUD细胞的数目、面积和平均灰度值;用原位杂交法检测大鼠胃窦平滑肌细胞中GASR及VIPR2 mRNA的阳性细胞表达情况。
结果:(1)一定剂量的甘草煎剂可延长大鼠的胃半排时间,两个模型组与正常组比较,有显著性差异(P<0.05或P<0.01)。模型3天组及模型7天组大鼠血液及胃窦组织中MTL、GAS及SP含量均降低,与正常组比较,有显著性差异(P<0.05或P<0.01);大鼠血液及胃窦组织中VIP及SS含量明显上升,与正常组比较,有显著性差异(P<0.05或P<0.01)。用一定剂量的甘草煎剂造模后,大鼠胃窦组织中的5-HT APUD细胞数目及面积明显减少、平均灰度值增高,两个模型组与正常组比较,有非常显著性差异(P<0.01)。模型3天组及模型7天组使有GASR
mRNA表达的阳性细胞的平均光密度明显降低,平均灰度值均升高,与正常组比较,有非常显著性差异(P<0.01);模型3天组及模型7天组使有VIPR2 mRNA表达的阳性细胞的平均光密度升高,平均灰度值明显降低,与正常组比较,有显著性差异(P<0.01)。
(2)旋复代赭汤各剂量组及吗丁啉组与两个模型组比较,可使血液及胃窦组织中MTL含量均增加,有显著性差异(P<0.05或P<0.01);旋复代赭汤中、高
剂量组与吗丁琳组比较,血液及组织中MTL含量增加明显,有显著性差异(P< 0.05
或P 均可使血液中GAs含量均增高,有显著性差异(P<0.肠或P<0.01);仅旋复代储
汤高剂量组可使组织中GAs含量明显增高,与模型3天组比较,有显著性差异
〔P 无显著性差异(P>0.05)。旋复代褚汤各剂量组及吗丁琳组与两个模型组比较,
血液及组织中SP含量增加,有显著性差异(P<0.05或P<0.时);旋复代储汤各
剂量组与吗丁淋组比较,除低剂量组对组织中SP含量的影响外,其余各组对大
鼠血液及组织中SP含量的影响均无显著性差异〔P>0.仍)。旋复代褚汤中、高剂
量组及吗丁琳组可降低胃动力低下大鼠血液中vIP含量,与模型3天组及模型7
天组比较,有显著性差异(P<0.肠或P<0.叭),其中高剂量组的作用与吗丁琳组
相当(P>0.05);各剂量组均可使胃动力低下大鼠组织中vIP含量降低,与两个
模型组比较,有显著性差异(P<0.仍或P (P>0.05)。旋复代赌汤各剂量组和吗丁琳组与两个模型组比较,血液中ss含量
明显降低,有显著性差异(P<0.似),三个剂量组的作用与吗丁琳组相当,无显
著性差异(P>0.05);旋复代储汤中、高剂量组及吗丁琳组与模型3天组比较,
组织中55含量降低,有显著性差异(P 与模型7天组比较,组织中ss含量降低,有显著性差异(P<0.05或P<0.01),
各剂量组对组织中SS含量的影响与吗丁琳组比较,无显著性差异(P>0.05)。
(3)旋复代褚汤中、高剂量组及吗丁琳组能明显升高胃动力低下大鼠胃窦
组织中5?
Objective: Gastrointestinal dismotility caused by many kinds of reasons is one group of disease as characteristic of delayed gastric emptying and hypoperistalsis of intestine, and it is common pathologic change of numerous functional digestive canal diseases. In addition, some organic digestive canal diseases, such as peptic ulcer, chronic gastritis and carcinoma of stomach , etc. , have gastrointestinal dismotility at the same time. In modern society, because the competition is aggravated , the rhythm of life is accelerated, people' s spirit is more and more tense , the excitability of the vagus nerve is strengthened ,which makes the emergence of disease of gastrointestinal dismotility rise. In recent years, according to the prevention and cure of gastrointestinal dismotility, the foreign scholars have launched the study on medicine of urging the power of intestines and stomach extensively. In the developed country, the costing for researching and developing new medicine of urging the power of intestines and stomach is up to 1 billion dollars equally every year. Some medicines, such as metoclopramide, domperidone, cisapride, etc., have already been applied at clinic. But curative effect and range of application of these medicines are still not ideal, and these medicines have lots of side effects to the central nervous system, circulatory system, and so on. Our country does not develop ideal medicines of urging the power of intestines and stomach either at present. It is discovered that XFDZT has remarkable curative effect to various kinds of disease of gastrointestinal dismotility. This subject makes the rat' s model of gastric dismotility with the liquorice decoction of certain dosage , in order to observe XFDZT ' influence on brain-gut peptides content and 5- HT APUD cell figure and brain-gut peptides receptor mRNA expressing in rat's blood or gastric antrum. This will offer the scientific theoretical foundation for widening XFDZT clinical practice and development of medicine.Methods: Firstly, made the rat's model of gastric dismotility with the liquorice decoction: 30 Wistar rats were divide into normal group, model group of 3 days and model group of 7 days at random, and every group had 10 rats (male and female rats were half and half). After giving liquorice decoction to rats, we tested rat' s stomach T1/2 with radioactive nuclide mark. Then 70 Wistar rats were divided into normal group(10 rats) and model group(60 rats) at random. Each rat of model group was irritated stomach with liquorice decoction(l0g/kg of weight). After irritated stomach at third day, 60 rats were divided into model group, model group of 3 days, model group of 7 days, low, hit, high dosage groups of XFDZT, group of domperidone .Except model group of 3 days, at the beginning of the fourth day, the rats of every other group were irritated stomach with liquorice decoction at 8:00a.m., and were cured 4:00p.m. Experiment was
finished at seventh day. Then tested brain-gut peptides content in blood and gastric antrum with radioimmunopercipitation assay and 5- HT APUD cell figure with immunohistochemical method and brain-gut peptides receptor mRNA express with orthotopic hybridization.Results: (1) Liquorice decoction of certain dosage could lengthen rat's stomach T1/2, and there was significance difference between model groups and normal group (P<0. 05 or P<0. 01). In rat's blood and organization of model groups, MTL、GAS and SP content were reduced , compared with the normal group, there was significance difference (P<0. 05 or P<0. 01 ) ; VIP and SS content rised obviously, compared with the normal group, there was significance difference (P<0. 05 or P<0. 01). 5-HT APUD cell figure and area reduced obviously, and there was extraordinary significance difference between model groups and normal group (P<0. 01). Average ligh density of cell that had GASR mRNA express obviously reduced, and average grey level rised, and there were extraordinary significance difference between model groups and normal group (P<0.01); Average high density of cell that had
方法:先用甘草煎剂制作胃动力低下模型:将Wistar大鼠30只,随机分为正常组、模型3天组及模型7天组,每组10只,雌雄各半。用一定剂量的甘草煎剂灌胃给药后,用放射性核素标记法检测大鼠的胃半排时间。再将健康Wistar大鼠70只随机分为正常组和模型组,正常组10只,模型组60只,雌雄各半。模型组每只大鼠按10g/kg体重的甘草煎剂灌胃,第三次给药前24小时动物禁食不禁水。第三天给药后,将60只大鼠随机分为模型3天组、模型7天组、旋复代赭汤低、中、高剂量组、吗丁啉组。每组10只,雌雄各半。除了模型3天组外,从第四天开始,模型7天组、旋复代赭汤各剂量组及吗丁啉组继续用甘草煎剂予上午8:00灌胃,直至第7天结束;从第三天开始,下午4:00正常组及模型7天组予等体积的生理盐水灌胃,吗丁啉组予0.27mg/ml的吗丁啉混悬液按100g体重1ml灌胃,中药低、中、高各剂量组分别予浓度为0.369g/ml、0.738g/ml、1.476g/ml的旋复代赭汤灌胃,共给药5天,末次给药前24小时动物禁食不禁水。用放免法观测各组大鼠血液及组织中脑肠肽的含量;用免疫组化法检测胃窦组织中5-HT APUD细胞的数目、面积和平均灰度值;用原位杂交法检测大鼠胃窦平滑肌细胞中GASR及VIPR2 mRNA的阳性细胞表达情况。
结果:(1)一定剂量的甘草煎剂可延长大鼠的胃半排时间,两个模型组与正常组比较,有显著性差异(P<0.05或P<0.01)。模型3天组及模型7天组大鼠血液及胃窦组织中MTL、GAS及SP含量均降低,与正常组比较,有显著性差异(P<0.05或P<0.01);大鼠血液及胃窦组织中VIP及SS含量明显上升,与正常组比较,有显著性差异(P<0.05或P<0.01)。用一定剂量的甘草煎剂造模后,大鼠胃窦组织中的5-HT APUD细胞数目及面积明显减少、平均灰度值增高,两个模型组与正常组比较,有非常显著性差异(P<0.01)。模型3天组及模型7天组使有GASR
mRNA表达的阳性细胞的平均光密度明显降低,平均灰度值均升高,与正常组比较,有非常显著性差异(P<0.01);模型3天组及模型7天组使有VIPR2 mRNA表达的阳性细胞的平均光密度升高,平均灰度值明显降低,与正常组比较,有显著性差异(P<0.01)。
(2)旋复代赭汤各剂量组及吗丁啉组与两个模型组比较,可使血液及胃窦组织中MTL含量均增加,有显著性差异(P<0.05或P<0.01);旋复代赭汤中、高
剂量组与吗丁琳组比较,血液及组织中MTL含量增加明显,有显著性差异(P< 0.05
或P
汤高剂量组可使组织中GAs含量明显增高,与模型3天组比较,有显著性差异
〔P
血液及组织中SP含量增加,有显著性差异(P<0.05或P<0.时);旋复代储汤各
剂量组与吗丁淋组比较,除低剂量组对组织中SP含量的影响外,其余各组对大
鼠血液及组织中SP含量的影响均无显著性差异〔P>0.仍)。旋复代褚汤中、高剂
量组及吗丁琳组可降低胃动力低下大鼠血液中vIP含量,与模型3天组及模型7
天组比较,有显著性差异(P<0.肠或P<0.叭),其中高剂量组的作用与吗丁琳组
相当(P>0.05);各剂量组均可使胃动力低下大鼠组织中vIP含量降低,与两个
模型组比较,有显著性差异(P<0.仍或P
明显降低,有显著性差异(P<0.似),三个剂量组的作用与吗丁琳组相当,无显
著性差异(P>0.05);旋复代储汤中、高剂量组及吗丁琳组与模型3天组比较,
组织中55含量降低,有显著性差异(P
各剂量组对组织中SS含量的影响与吗丁琳组比较,无显著性差异(P>0.05)。
(3)旋复代褚汤中、高剂量组及吗丁琳组能明显升高胃动力低下大鼠胃窦
组织中5?
Objective: Gastrointestinal dismotility caused by many kinds of reasons is one group of disease as characteristic of delayed gastric emptying and hypoperistalsis of intestine, and it is common pathologic change of numerous functional digestive canal diseases. In addition, some organic digestive canal diseases, such as peptic ulcer, chronic gastritis and carcinoma of stomach , etc. , have gastrointestinal dismotility at the same time. In modern society, because the competition is aggravated , the rhythm of life is accelerated, people' s spirit is more and more tense , the excitability of the vagus nerve is strengthened ,which makes the emergence of disease of gastrointestinal dismotility rise. In recent years, according to the prevention and cure of gastrointestinal dismotility, the foreign scholars have launched the study on medicine of urging the power of intestines and stomach extensively. In the developed country, the costing for researching and developing new medicine of urging the power of intestines and stomach is up to 1 billion dollars equally every year. Some medicines, such as metoclopramide, domperidone, cisapride, etc., have already been applied at clinic. But curative effect and range of application of these medicines are still not ideal, and these medicines have lots of side effects to the central nervous system, circulatory system, and so on. Our country does not develop ideal medicines of urging the power of intestines and stomach either at present. It is discovered that XFDZT has remarkable curative effect to various kinds of disease of gastrointestinal dismotility. This subject makes the rat' s model of gastric dismotility with the liquorice decoction of certain dosage , in order to observe XFDZT ' influence on brain-gut peptides content and 5- HT APUD cell figure and brain-gut peptides receptor mRNA expressing in rat's blood or gastric antrum. This will offer the scientific theoretical foundation for widening XFDZT clinical practice and development of medicine.Methods: Firstly, made the rat's model of gastric dismotility with the liquorice decoction: 30 Wistar rats were divide into normal group, model group of 3 days and model group of 7 days at random, and every group had 10 rats (male and female rats were half and half). After giving liquorice decoction to rats, we tested rat' s stomach T1/2 with radioactive nuclide mark. Then 70 Wistar rats were divided into normal group(10 rats) and model group(60 rats) at random. Each rat of model group was irritated stomach with liquorice decoction(l0g/kg of weight). After irritated stomach at third day, 60 rats were divided into model group, model group of 3 days, model group of 7 days, low, hit, high dosage groups of XFDZT, group of domperidone .Except model group of 3 days, at the beginning of the fourth day, the rats of every other group were irritated stomach with liquorice decoction at 8:00a.m., and were cured 4:00p.m. Experiment was
finished at seventh day. Then tested brain-gut peptides content in blood and gastric antrum with radioimmunopercipitation assay and 5- HT APUD cell figure with immunohistochemical method and brain-gut peptides receptor mRNA express with orthotopic hybridization.Results: (1) Liquorice decoction of certain dosage could lengthen rat's stomach T1/2, and there was significance difference between model groups and normal group (P<0. 05 or P<0. 01). In rat's blood and organization of model groups, MTL、GAS and SP content were reduced , compared with the normal group, there was significance difference (P<0. 05 or P<0. 01 ) ; VIP and SS content rised obviously, compared with the normal group, there was significance difference (P<0. 05 or P<0. 01). 5-HT APUD cell figure and area reduced obviously, and there was extraordinary significance difference between model groups and normal group (P<0. 01). Average ligh density of cell that had GASR mRNA express obviously reduced, and average grey level rised, and there were extraordinary significance difference between model groups and normal group (P<0.01); Average high density of cell that had
引文
1 黄兆胜.中药学.第一版.北京:人民卫生出版社,2002:426-427
2 中国药典委员会.中国药典.第一部.北京:科学出版社,1977:3
3 崔志清,郭世铎,张惠茹等.炙甘草和甘草酸对翻转小鼠离体小肠糖-钠转运电位的影响.中草药.1995;26(12):643
4 王访,苏耀海.甘草的药理作用及临床应用.时珍国医国药.2002;13(5):303-304
5 李铁民,梁再赋.甘草提取物及其衍生物的抗病毒研究现状.中草药.1994;25 (12):657
6 陈晓光,韩锐.甘草次酸对苯并芘诱发DNA损伤及非程序DNA合成的影响.药学学报.1994;29(10):725
7 严瑞琪,李俊丽,黄玫玲等.甘草甜素抑制致癌过程中对DNA损伤修复的影响.癌症.1995;14(4):245
8 张萍,祝稀娴.甘草及其制剂药理与临床应用研究新进展.中草药.1997;28 (9):568-569
9 税典奎,姜楠,刘松林.不同剂量的甘草煎剂对小鼠胃肠运动的影响.江苏中 医药.2005;26(5):53-54
10 朱欣佚,窦国祥,杨德治.通腑汤对胃肠动力影响的实验研究.南京铁道医学院学报.2000;19(4):244
11 王长洪,陈多,等.旋复代赭汤促胃肠动力作用的实验研究.中国中西医结合脾胃杂志.1999;7(1):6-7
12 陈多,吴春福,等.旋复代赭汤促胃肠动力作用研究Ⅰ.对小鼠胃排空的影响.中药药理与临床.1997;13(1):4-5
13 陈多,吴春福,等.旋复代赭汤促胃肠动力作用研究Ⅲ.旋复代赭汤对小鼠小肠推进作用的影响.中药药理与临床.1997;13(4):7-8
14 陈多,吴春福,等.旋复代赭汤促胃肠动力作用研究Ⅳ.全方与拆方对在体胃和离体胃作用的异同.中药药理与临床.1998;14(5):1-3
15 吴春福.旋复代赭汤和半夏泻心汤促胃肠动力作用研究.哈尔滨商业大学学报.2002;18(1):6-7
16 张燕,刘俊田,等.胃肠舒泰颗粒对大鼠胃泌素含量及小鼠胃肠运动功能的影响.中国中西医结合消化杂志.2002;10(6):333-336
17 任平,刘芳,等.脾虚大鼠生长抑素、胃动素、胆囊收缩素内分泌细胞的变化.成都中医药大学学报.2001;24(1):35-37
18 任平,黄熙,等.四君子汤对实验性脾虚大鼠胃排空率的影响.中国中西医结合杂志.2000;20(8):596-598
19 周吕,柯美云.胃肠动力学.第一版.北京:科学出版社,1999:400
2 中国药典委员会.中国药典.第一部.北京:科学出版社,1977:3
3 崔志清,郭世铎,张惠茹等.炙甘草和甘草酸对翻转小鼠离体小肠糖-钠转运电位的影响.中草药.1995;26(12):643
4 王访,苏耀海.甘草的药理作用及临床应用.时珍国医国药.2002;13(5):303-304
5 李铁民,梁再赋.甘草提取物及其衍生物的抗病毒研究现状.中草药.1994;25 (12):657
6 陈晓光,韩锐.甘草次酸对苯并芘诱发DNA损伤及非程序DNA合成的影响.药学学报.1994;29(10):725
7 严瑞琪,李俊丽,黄玫玲等.甘草甜素抑制致癌过程中对DNA损伤修复的影响.癌症.1995;14(4):245
8 张萍,祝稀娴.甘草及其制剂药理与临床应用研究新进展.中草药.1997;28 (9):568-569
9 税典奎,姜楠,刘松林.不同剂量的甘草煎剂对小鼠胃肠运动的影响.江苏中 医药.2005;26(5):53-54
10 朱欣佚,窦国祥,杨德治.通腑汤对胃肠动力影响的实验研究.南京铁道医学院学报.2000;19(4):244
11 王长洪,陈多,等.旋复代赭汤促胃肠动力作用的实验研究.中国中西医结合脾胃杂志.1999;7(1):6-7
12 陈多,吴春福,等.旋复代赭汤促胃肠动力作用研究Ⅰ.对小鼠胃排空的影响.中药药理与临床.1997;13(1):4-5
13 陈多,吴春福,等.旋复代赭汤促胃肠动力作用研究Ⅲ.旋复代赭汤对小鼠小肠推进作用的影响.中药药理与临床.1997;13(4):7-8
14 陈多,吴春福,等.旋复代赭汤促胃肠动力作用研究Ⅳ.全方与拆方对在体胃和离体胃作用的异同.中药药理与临床.1998;14(5):1-3
15 吴春福.旋复代赭汤和半夏泻心汤促胃肠动力作用研究.哈尔滨商业大学学报.2002;18(1):6-7
16 张燕,刘俊田,等.胃肠舒泰颗粒对大鼠胃泌素含量及小鼠胃肠运动功能的影响.中国中西医结合消化杂志.2002;10(6):333-336
17 任平,刘芳,等.脾虚大鼠生长抑素、胃动素、胆囊收缩素内分泌细胞的变化.成都中医药大学学报.2001;24(1):35-37
18 任平,黄熙,等.四君子汤对实验性脾虚大鼠胃排空率的影响.中国中西医结合杂志.2000;20(8):596-598
19 周吕,柯美云.胃肠动力学.第一版.北京:科学出版社,1999:400