柴胡枳实提取物促胃肠动力作用的实验研究
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摘要
目的
比较柴胡枳实水提物、醇提物、醇水提物促胃肠动力作用，筛选最优提取物，观察其对阿托品、吗啡引起胃肠抑制模型的作用以及血浆胃动素、不同组织一氧化氮(NO)含量的影响，探讨其促胃肠动力作用的机制。
方法
1．选取北柴胡、酸橙枳实分别制备水提物、醇提物、醇水提物。
2．昆明小鼠随机分成11组，每组20只，阴性对照组为蒸馏水组，阳性对照组为西沙比利组，余为不同剂量的各提取物组，用药三天，禁食12小时后，采用葡聚糖蓝2000水溶液为标记物，以相对胃内色素残留率及平均小肠推进比作为指标，比较不同剂量各提取物促胃肠动力的作用，筛选最优提取物。
3．昆明小鼠随机分成6组，每组15只，分别为A(蒸馏水)组、B(阿托品)组、D(吗啡)组、F(3倍常用量的水提取物)组、C(阿托品+3倍常用量的水提取物)组、E(吗啡+3倍常用量的水提取物)组，用药三天，禁食12小时后，于第四天灌服药物的同时B组、C组腹腔注射阿托品0.0003g／kg，E组、D组腹腔注射盐酸吗啡0.0003g／kg，继续应用上述标记物和指标，观察3倍常用量的水提取物对阿托品及吗啡引起胃肠抑制模型的影响。
4．Wistar大鼠随机分成二组，一组为中药组，另一组为正常对照组，每组15只，用药六天，禁食12小时后，以葡聚糖蓝2000水溶液为标记物，测大鼠相对胃内色素残留率及平均小肠推进比的同时采用放射免疫法测定血浆胃动素浓度、采用硝酸还原酶法测定其组织NO的含量。
5．数据处理：应用SPSS10.0软件进行统计学处理，各数据用均数±标准差表示，两组间比较用t检验。
结果
1.醇提物14mg/g组相对胃内色素残留率及平均小肠推进比与蒸馏水
组相比无显著性差异(P>0.05)，余提取物组与蒸馏水组相比具有非常显
著性差异(P<0.01)或显著性差异(P<0.05)，其中3倍常用量的水提取物
4.Zm扩g组相对胃内色素残留率明显降低、平均小肠推进比明显升高，其与
西沙比利组相比无显著性差异(P>0.05)。
2.分别比较二组间的相对胃内色素残留率及平均小肠推进比，B(阿托
品组与A(蒸馏水)组、D(吗啡)组与A(蒸馏水)组、D(吗啡)组与E(吗啡
十3倍常用量的水提取物)组、E(吗啡+3倍常用量的水提取物)组与F(3
倍常用量的水提取物)组、C(阿托品+3倍常用量的水提取物)组与F(3倍
常用量的水提取物)组相比均具有显著性差异(P<0.05)，而B(阿托品)组
与C(阿托品+3倍常用量的水提取物)组相比无显著性差异(P>0.05)。
3.H(3倍常用量的水提取物)组相对胃内色素残留率及平均小肠推进
比与G(蒸馏水)组相比具有非常显著性差异(P<0.01);H组血浆胃动素
与G组相比具有显著性差异(P<0.05);H组胃底、胃窦、空肠NO含量与
G组相比具有非常显著性差异(P<0.01)。
结论
1.柴胡积实水提物3倍常用量为最优提取物;
2.3倍常用量的水提取物不能拮抗阿托品的作用，能够拮抗吗啡所致
的胃肠抑制运动;
3.3倍常用量的水提取物能够明显提高血浆胃动素的水平，同时使胃
底组织NO含量显著增加，胃窦及空肠组织NO含量显著减少而促进胃肠
运动。
Objective
To compare the stimulative effect on gastrointestinal motility of water extraction ^ethanol extraction and ethanol water mixed extraction of Bupleurum and Citrus to screen out the optimum extraction (TOE) , to observe that it influenced on the gastrointestinal inhibitive model induced by Atropin and Morphine and the contents of nitric oxide (NO) in the gastric fundusN gastric antrum and jejunum tissue in order to explore the mechanism of the stimulative effect on gastrointestinal motility .
Methods
1. To select B. Scorzonerifolium Willd and Citrus aurantium to execute water extraction ,ethanol extraction and ethanol water mixed extraction respectively.
2. Kunming mice were randomly divided into 11 groups,each group was 20 mice, negative control was distilled water group, positive control was Cisapride group, the rest were different dosage extraction groups, compared the stimulative effect of different dosage extractions on gastrointestinal motility with Blue Dxtran 2000 as a marker ,by relative pigment remainder rate inside stomach and average small intestinal propulsive ratio as guide lines.
3. Kunming mice were randomly divided into 6 groups, each group was 15 mice ,they were distilled water group ,Atropin group ,Morphine group , water extraction of 3 times usual dosage (3WEOTUD) group , Morphine and 3WEOTUD group , Atropin and 3WEOTUD group respectively. To continue applying the
above the marker and guide lines to observe that 3WEOTUD influenced on the gastrointestinal inhibitive model induced by Atropin and Morphine.
4. Wistar rats were randomly divided into 2 groups,one group was Chinese traditional medicine group, another was normal control group ,each group was 15 rats. The contents of nitric oxide (NO) in the gastric fundus^ gastric antrum and jejunum tissue were measured with the method of nitrate reductase at the same time ,the plasma motilin(MTL) level was detected with radioimmunoassay .
5. Statistical analysis: All date were expressedas Mean SD and analyzed between two groups by T test with statistical software for SPSS 10.0 .
Results
1. Every extraction group except the ethanol extraction 14mg/g group compared with distilled water group, it was highly significant difference( P < 0. 01) or significant difference( P < 0. 05) , the water extraction of 3 times usual dosage compared with Cisapride group , it wasn't significant difference.
2. To compared relative pigment remainder rate inside stomach and average small intestinal propulsive ratio respectively between two groups, B group comparing with A group ,D group comparing with A group ,D group comparing with E group ,E group comparing with F group and C group comparing with F Group were significant difference ( P < 0. 05) , but B group comparing with C group wasn' t significant difference( P > 0. 05 ).
3. H group compared with G group on relative pigment remainder rate inside stomach and average small intestinal propulsive ratio, it was highly significant difference (P<0.01), H group compared with G group in plasma motilin, it was significant difference (P <0. 05) , the content of NO in the gastric fundus, gastric antrum and jejunum of H group compared with G group was highly significant difference( P < 0.01) .
Conclusion
1. 3WEOTUD was confirmed the optimum extraction .
2. 3WEOTUD couldn't antagonize the gastrointestinal inhibitive effect induced by Atropin , but it could antagonize gastrointestinal inhibiting effect induced by Morphine.
3.3WEOTUD could promote gastrointestinal motility of rats obviously and enhance plasma motilin level ,the content of NO in the gastric fundus tissue of rats was increased significantly , but the content of NO in the gastric antrum and jejunum tissue were decreased significantly.
比较柴胡枳实水提物、醇提物、醇水提物促胃肠动力作用，筛选最优提取物，观察其对阿托品、吗啡引起胃肠抑制模型的作用以及血浆胃动素、不同组织一氧化氮(NO)含量的影响，探讨其促胃肠动力作用的机制。
方法
1．选取北柴胡、酸橙枳实分别制备水提物、醇提物、醇水提物。
2．昆明小鼠随机分成11组，每组20只，阴性对照组为蒸馏水组，阳性对照组为西沙比利组，余为不同剂量的各提取物组，用药三天，禁食12小时后，采用葡聚糖蓝2000水溶液为标记物，以相对胃内色素残留率及平均小肠推进比作为指标，比较不同剂量各提取物促胃肠动力的作用，筛选最优提取物。
3．昆明小鼠随机分成6组，每组15只，分别为A(蒸馏水)组、B(阿托品)组、D(吗啡)组、F(3倍常用量的水提取物)组、C(阿托品+3倍常用量的水提取物)组、E(吗啡+3倍常用量的水提取物)组，用药三天，禁食12小时后，于第四天灌服药物的同时B组、C组腹腔注射阿托品0.0003g／kg，E组、D组腹腔注射盐酸吗啡0.0003g／kg，继续应用上述标记物和指标，观察3倍常用量的水提取物对阿托品及吗啡引起胃肠抑制模型的影响。
4．Wistar大鼠随机分成二组，一组为中药组，另一组为正常对照组，每组15只，用药六天，禁食12小时后，以葡聚糖蓝2000水溶液为标记物，测大鼠相对胃内色素残留率及平均小肠推进比的同时采用放射免疫法测定血浆胃动素浓度、采用硝酸还原酶法测定其组织NO的含量。
5．数据处理：应用SPSS10.0软件进行统计学处理，各数据用均数±标准差表示，两组间比较用t检验。
结果
1.醇提物14mg/g组相对胃内色素残留率及平均小肠推进比与蒸馏水
组相比无显著性差异(P>0.05)，余提取物组与蒸馏水组相比具有非常显
著性差异(P<0.01)或显著性差异(P<0.05)，其中3倍常用量的水提取物
4.Zm扩g组相对胃内色素残留率明显降低、平均小肠推进比明显升高，其与
西沙比利组相比无显著性差异(P>0.05)。
2.分别比较二组间的相对胃内色素残留率及平均小肠推进比，B(阿托
品组与A(蒸馏水)组、D(吗啡)组与A(蒸馏水)组、D(吗啡)组与E(吗啡
十3倍常用量的水提取物)组、E(吗啡+3倍常用量的水提取物)组与F(3
倍常用量的水提取物)组、C(阿托品+3倍常用量的水提取物)组与F(3倍
常用量的水提取物)组相比均具有显著性差异(P<0.05)，而B(阿托品)组
与C(阿托品+3倍常用量的水提取物)组相比无显著性差异(P>0.05)。
3.H(3倍常用量的水提取物)组相对胃内色素残留率及平均小肠推进
比与G(蒸馏水)组相比具有非常显著性差异(P<0.01);H组血浆胃动素
与G组相比具有显著性差异(P<0.05);H组胃底、胃窦、空肠NO含量与
G组相比具有非常显著性差异(P<0.01)。
结论
1.柴胡积实水提物3倍常用量为最优提取物;
2.3倍常用量的水提取物不能拮抗阿托品的作用，能够拮抗吗啡所致
的胃肠抑制运动;
3.3倍常用量的水提取物能够明显提高血浆胃动素的水平，同时使胃
底组织NO含量显著增加，胃窦及空肠组织NO含量显著减少而促进胃肠
运动。
Objective
To compare the stimulative effect on gastrointestinal motility of water extraction ^ethanol extraction and ethanol water mixed extraction of Bupleurum and Citrus to screen out the optimum extraction (TOE) , to observe that it influenced on the gastrointestinal inhibitive model induced by Atropin and Morphine and the contents of nitric oxide (NO) in the gastric fundusN gastric antrum and jejunum tissue in order to explore the mechanism of the stimulative effect on gastrointestinal motility .
Methods
1. To select B. Scorzonerifolium Willd and Citrus aurantium to execute water extraction ,ethanol extraction and ethanol water mixed extraction respectively.
2. Kunming mice were randomly divided into 11 groups,each group was 20 mice, negative control was distilled water group, positive control was Cisapride group, the rest were different dosage extraction groups, compared the stimulative effect of different dosage extractions on gastrointestinal motility with Blue Dxtran 2000 as a marker ,by relative pigment remainder rate inside stomach and average small intestinal propulsive ratio as guide lines.
3. Kunming mice were randomly divided into 6 groups, each group was 15 mice ,they were distilled water group ,Atropin group ,Morphine group , water extraction of 3 times usual dosage (3WEOTUD) group , Morphine and 3WEOTUD group , Atropin and 3WEOTUD group respectively. To continue applying the
above the marker and guide lines to observe that 3WEOTUD influenced on the gastrointestinal inhibitive model induced by Atropin and Morphine.
4. Wistar rats were randomly divided into 2 groups,one group was Chinese traditional medicine group, another was normal control group ,each group was 15 rats. The contents of nitric oxide (NO) in the gastric fundus^ gastric antrum and jejunum tissue were measured with the method of nitrate reductase at the same time ,the plasma motilin(MTL) level was detected with radioimmunoassay .
5. Statistical analysis: All date were expressedas Mean SD and analyzed between two groups by T test with statistical software for SPSS 10.0 .
Results
1. Every extraction group except the ethanol extraction 14mg/g group compared with distilled water group, it was highly significant difference( P < 0. 01) or significant difference( P < 0. 05) , the water extraction of 3 times usual dosage compared with Cisapride group , it wasn't significant difference.
2. To compared relative pigment remainder rate inside stomach and average small intestinal propulsive ratio respectively between two groups, B group comparing with A group ,D group comparing with A group ,D group comparing with E group ,E group comparing with F group and C group comparing with F Group were significant difference ( P < 0. 05) , but B group comparing with C group wasn' t significant difference( P > 0. 05 ).
3. H group compared with G group on relative pigment remainder rate inside stomach and average small intestinal propulsive ratio, it was highly significant difference (P<0.01), H group compared with G group in plasma motilin, it was significant difference (P <0. 05) , the content of NO in the gastric fundus, gastric antrum and jejunum of H group compared with G group was highly significant difference( P < 0.01) .
Conclusion
1. 3WEOTUD was confirmed the optimum extraction .
2. 3WEOTUD couldn't antagonize the gastrointestinal inhibitive effect induced by Atropin , but it could antagonize gastrointestinal inhibiting effect induced by Morphine.
3.3WEOTUD could promote gastrointestinal motility of rats obviously and enhance plasma motilin level ,the content of NO in the gastric fundus tissue of rats was increased significantly , but the content of NO in the gastric antrum and jejunum tissue were decreased significantly.
引文
1．李岩，陈苏宁，李宇权，等．芍药甘草汤、四逆散对胃排空及小肠推进功能影响的拆方研究．中华消化杂志，1996；16(1)：18—21
2．李岩，陈苏宁，陈少夫，等．柴胡枳实合剂对健康人胃排空、小肠推进功能及血中胃动素、胃泌素水平的影响．中国中西医结合杂志，1996；16(10)：622
3．郝庆，李岩，尹洪涛．不同品种柴胡枳实对小鼠胃肠动力作用的比较研究．华人消化杂志，1998；6(特刊7)：205-207
4．郝庆，李岩，张桓军．柴胡枳实促胃肠动力作用机制的实验研究．辽宁药物与临床，2002；5(2)：65-66
5．田丰，沈静雪，王伟，等．柴胡枳实合剂对运动障碍样型功能性消化不良患者症状、胃排空、胃电图的影响．世界华人消化杂志，2002；10(11)：1357-1359
6．谢秀琼主编．中药新药制剂开发与应用．第二版．人民卫生出版社，2002；124-125
7．陈奇主编．中药药理研究方法学．第一版．人民卫生出版社，1993；1103
8．李岩，陈苏宁，田代真一．葡聚糖蓝2000标记五积散对小鼠胃排空及小肠推进功能的影响．中国医科大学学报，1996；25(2)：160-161
9. Iwanaga Y, Miyashita N, Mizutani F, et al. Stimulatory effect of N - [4 - [2- (Di - methylamino) - ethoxy ] benzyl ] - 3,4 - dimethoxybenzamide hydro -chloride (HSR-803)on normal and delayed gastrointestinal propulsion. Japan J Pharmacol, 1991; 56: 261
10．彭国平，牛贺明，徐丽华．枳实活性成分的研究．南京中医药大学学报(自然科学版)，2001；17(2)：91-92
11．胡盛珊，王大元，邱萍．枳实有效成分的药理活性比较．中草药，1994；25(8)：419-421
12．朱金照，杨国汉，冷恩仁．中药的促胃肠动力作用．世界华人消化杂志，1999；7(8)：687-688
13．陈多，吴春福，王长洪，刘雯．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的影响．中药药理与临床，1997；13(4)：7-9
14．黄颖秋．胃轻瘫综合征的中医中药治疗．世界华人消化杂志，2001；9(10)：1167—1168
15．黄子荷，杨德治，魏义全．阿托品对枳实增强犬小肠电活动的影响．中国中西医结合杂志，1996；16：292-293
16．马清钧，王淑玲主编．常用中药现代研究与临床[M]．第一版．天津科技翻译出版公司出版，1995；49
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24. Fiorucci S, Distrutti E, Quuintieri A. L- Arginine/nitric oxide pathway modulates gastric motility and gallbladder emptying induced by erythromycin and liquid meal in humans. Dig Dis Sci, 1995; 40: 1365 - 1371.
25. Kenton MS and Scan MW. Nitric oxide as a mediator of nonadrenergic neurotransmission. Am J Physiol, 1992; 263: G379
26．吴建维，蒋予，罗金燕，等．一氧化氮对小肠消化间期移行性复合运动作用的研究．深圳医学，1999；12(1)：17
2．李岩，陈苏宁，陈少夫，等．柴胡枳实合剂对健康人胃排空、小肠推进功能及血中胃动素、胃泌素水平的影响．中国中西医结合杂志，1996；16(10)：622
3．郝庆，李岩，尹洪涛．不同品种柴胡枳实对小鼠胃肠动力作用的比较研究．华人消化杂志，1998；6(特刊7)：205-207
4．郝庆，李岩，张桓军．柴胡枳实促胃肠动力作用机制的实验研究．辽宁药物与临床，2002；5(2)：65-66
5．田丰，沈静雪，王伟，等．柴胡枳实合剂对运动障碍样型功能性消化不良患者症状、胃排空、胃电图的影响．世界华人消化杂志，2002；10(11)：1357-1359
6．谢秀琼主编．中药新药制剂开发与应用．第二版．人民卫生出版社，2002；124-125
7．陈奇主编．中药药理研究方法学．第一版．人民卫生出版社，1993；1103
8．李岩，陈苏宁，田代真一．葡聚糖蓝2000标记五积散对小鼠胃排空及小肠推进功能的影响．中国医科大学学报，1996；25(2)：160-161
9. Iwanaga Y, Miyashita N, Mizutani F, et al. Stimulatory effect of N - [4 - [2- (Di - methylamino) - ethoxy ] benzyl ] - 3,4 - dimethoxybenzamide hydro -chloride (HSR-803)on normal and delayed gastrointestinal propulsion. Japan J Pharmacol, 1991; 56: 261
10．彭国平，牛贺明，徐丽华．枳实活性成分的研究．南京中医药大学学报(自然科学版)，2001；17(2)：91-92
11．胡盛珊，王大元，邱萍．枳实有效成分的药理活性比较．中草药，1994；25(8)：419-421
12．朱金照，杨国汉，冷恩仁．中药的促胃肠动力作用．世界华人消化杂志，1999；7(8)：687-688
13．陈多，吴春福，王长洪，刘雯．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的影响．中药药理与临床，1997；13(4)：7-9
14．黄颖秋．胃轻瘫综合征的中医中药治疗．世界华人消化杂志，2001；9(10)：1167—1168
15．黄子荷，杨德治，魏义全．阿托品对枳实增强犬小肠电活动的影响．中国中西医结合杂志，1996；16：292-293
16．马清钧，王淑玲主编．常用中药现代研究与临床[M]．第一版．天津科技翻译出版公司出版，1995；49
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