“乐胃饮”对实验性脾虚大鼠胃肠激素影响的研究
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摘要
目的 本实验用具有苦寒攻下作用的大黄塑造大鼠胃肠功能紊乱的脾虚模型，观察乐胃饮对大鼠血清中胃泌素(Gastrin，Gas)和血浆中胃动素(Motilin，Mot)含量的影响及对大鼠胃粘膜细胞形态学的改变，探讨中药复方乐胃饮-调整胃肠素-调控胃肠动力三者之间的关系及对胃粘膜细胞的保护作用，阐明乐胃饮治疗胃肠动力紊乱性疾病的机理。
方法 随机将70只大鼠分为正常组10只，实验组60只。实验组大鼠用200％(即1ml含大黄2克)大黄喂饲SD大鼠2周，造成胃肠功能紊乱的脾虚模型，后再用高、中、低三种剂量的乐胃饮灌服大鼠，同时用中成药香砂六君丸，西药西沙必利作阳性对照。给药2周后，摘眼球取血，用于测定血清胃泌素(GAS)和血浆胃动素(MOT)。然后断头处死动物，取出胃组织块，放入的100mL．L~(-1)福尔马林液中固定，制成切片以备观测胃粘膜形态学的改变。运用以上指标分别从胃肠激素水平和病理两个方面研究经验方“乐胃饮”治疗胃肠功能紊乱性疾病的治疗机理。
结果 ①实验鼠大体情况：在实验过程中，正常组大鼠饮食、活动、大便及毛发等均无异常，体重逐渐增加。而大黄造模1周后的大鼠出现活动减少，毛发枯槁(背脊毛色从白到红再转黄)，弓背，体重明显下降，食量减少，喜扎堆，大便稀溏，造模1周半后出现肛门脱垂，大便粘稠或成形软便。而中西药治疗组较模型组有不同程度的恢复。②胃肠激
素测定:a.血清胃泌素:模型组一与正常组比较有非常显著差
异，说明脾虚大鼠存在血清胃泌素(GAS)水平的显著阁氏;
中成药与西药组与模型组差异显著。乐胃饮高、中剂量组与
模型组差异亦显著，高剂量与模型组比只0.01，中剂量组与
模型组相比只0.05，从均数来看高剂量更结近正常组水平。
b.血浆胃动素:模型组与正常组相比较只0.01有非常显著差
异，说明脾虚大鼠存在血浆胃动素(M汀l…)水平的显著升高。
中成药与模型组相比有非常显著差异只0.01。乐胃饮高、中
剂量与模型组相比有显著差异只0 .05，而乐胃饮低剂量与正
常组有显著差异只0 .05，而其与模型组相比差异不显著。③
病理检测:10 x10低倍镜下观察各组切片未见淋巴细胞、浆
细胞或白细胞等炎性浸润，大黄诱发的脾虚模型组胃粘膜表
面上皮(柱状细胞)与胃底腺上皮细胞排列紊乱。10 X40高
倍镜下脾虚模型组能见到一些胃粘膜细胞的损伤，尤对壁细
胞明显，如细胞皱缩，肪明莫出现缺损和胞质嗜酸性较正常组
降低，颜色由红色转为暗红色，甚至有些壁细胞呈现空泡化，
核浓密，而中药乐胃饮和香砂六君丸、西沙必利5组的壁细
胞损伤有不同程度的恢复，细胞增大，胞质嗜酸性降低。
结论“乐胃饮”具有使异常改变的胃肠激素Gas和枷t
的含量恢复至正常水平，并且对脾虚大鼠相关胃粘膜的病理
改变有促进其恢复作用。从而表明经验方“乐胃饮”作用机
理之一可能是通过对胃肠激素水平的影响从而能够达到有效
的治疗胃肠动力紊乱性疾病的目的。
Objective We use Chinese herb "Da huang" with bitter-cold property to creati the spleen-deficient model which is of Gastro-intestinal dysfunction in the light of west medicine, observing"Le Wei Yin'Y influence on serum Gastrin content and plasm Motilin content in rat and morphological chang in the gastral mucosal cell; investigating the relationship among'Te Wei Yin"-gut homrone-gastralintestinal dynamical and its protective function to gastral mucosa; explaining action mechanism of "Le Wei Yu"Twhich is treated gsatral-intestinal dynamical disorder disease.
Methods Seventy rats were divided at random into two big groups: normal group(ten) and experimental group (sixty). We feed experimental rats on 200% Da huang decotion After two weeks, we create sucesessfuly spleen-deficient model.Then We picked tats eyeballs to collect blood, measuring respectively Gastrin, Motilin, which are used as two indexes for our research on the mechanisum of treatment of "Le Wei Yin"at the level of gastral-intestinal hormone. After that, we kill the rats by means of decollation, pick out stomach tissu, put into 100ml L formalin promptly to fix tie tissue, then dehydration by alclhol, embed by paraffin, staining by Hematoxylin-esoin(HE), make sections in order to investigate morphological chang in gastral mucosal cell.
Results (1)General state: During the experiment, rats of normal group have none abnormal phenomenon in the light of its appetite, activity, stool and
skin What's more, they all put up little by little. By contrast, rats of model group are withered and yellow, arch its back, losing its weight, poor appetite, loose stool, avertion to cold after one week. What's worse, after half and a week, rats got annus proplapse, stiff stool or loose stool but remaining its formation. (2)Gastral-intestinal hormone: a. Serum Gastrin content: There is a remarkablely difference (P<0.01) between modle group and control group which note that spllen-deficient rats 's serum Gastrin content is significantly low; There is a remarkablely difference between high, moderat dosage of "Le Wei Yin" and model groupHigh dosage compared with model group(P<0.01), moderate dosage compared with model group(P<0.05).From the angle of average, high dosage group is by far closer to normal group, b. Plasm Motilin content There is a remarkablely difference (P<0.01) between modle group and control group which note that spllen-deficient rats 's plasm Motilin content is significantly highPatent Chinese herb "Xiang sha liu jun wan" group is significantly differente from model group(P<0.01). High, moderate dosage of "Le Wei Yin" group possess remarkabley difference compared with model group (P<0.05), while low dosage of "Le Wei Yin" is significantly differente from normal group,and is not significantly differente from model group. (3)Under the inverted microscope: Through 100 times power lens, we observed sections of each group and did not find inflammatory infiltration such as lymphocyte, plasmablast etc. In spleen-deficient model group induced by "Da huang", We find superficial epithelial cell and fundic gland epithelial cell of gastral mucosal cell arranging disorder. Through 400 times high power lens, We find some gastral mucosal cell damage, espetially
the parietal cell, while that in "Le Wei Yin", "Xiang sha liu jun", "Xi sha bi li" groups, cellular damage resume to some degree and some of them get large with cytoplast acidophilic decreasing.
Conclusions From above, experierre prescription''Le Wei Yin" possess the function of changing abnormal gut-hormon GAS and MOT content into normal state and the function of resuming gastral mucosal pathological change of spleen-deficient rats. So it mate clear that influence on gut-hormon may be one of action mechanism of "Le Wei Yin"efrective treatment of gut-intestinal dynamical disorder disease.
方法 随机将70只大鼠分为正常组10只，实验组60只。实验组大鼠用200％(即1ml含大黄2克)大黄喂饲SD大鼠2周，造成胃肠功能紊乱的脾虚模型，后再用高、中、低三种剂量的乐胃饮灌服大鼠，同时用中成药香砂六君丸，西药西沙必利作阳性对照。给药2周后，摘眼球取血，用于测定血清胃泌素(GAS)和血浆胃动素(MOT)。然后断头处死动物，取出胃组织块，放入的100mL．L~(-1)福尔马林液中固定，制成切片以备观测胃粘膜形态学的改变。运用以上指标分别从胃肠激素水平和病理两个方面研究经验方“乐胃饮”治疗胃肠功能紊乱性疾病的治疗机理。
结果 ①实验鼠大体情况：在实验过程中，正常组大鼠饮食、活动、大便及毛发等均无异常，体重逐渐增加。而大黄造模1周后的大鼠出现活动减少，毛发枯槁(背脊毛色从白到红再转黄)，弓背，体重明显下降，食量减少，喜扎堆，大便稀溏，造模1周半后出现肛门脱垂，大便粘稠或成形软便。而中西药治疗组较模型组有不同程度的恢复。②胃肠激
素测定:a.血清胃泌素:模型组一与正常组比较有非常显著差
异，说明脾虚大鼠存在血清胃泌素(GAS)水平的显著阁氏;
中成药与西药组与模型组差异显著。乐胃饮高、中剂量组与
模型组差异亦显著，高剂量与模型组比只0.01，中剂量组与
模型组相比只0.05，从均数来看高剂量更结近正常组水平。
b.血浆胃动素:模型组与正常组相比较只0.01有非常显著差
异，说明脾虚大鼠存在血浆胃动素(M汀l…)水平的显著升高。
中成药与模型组相比有非常显著差异只0.01。乐胃饮高、中
剂量与模型组相比有显著差异只0 .05，而乐胃饮低剂量与正
常组有显著差异只0 .05，而其与模型组相比差异不显著。③
病理检测:10 x10低倍镜下观察各组切片未见淋巴细胞、浆
细胞或白细胞等炎性浸润，大黄诱发的脾虚模型组胃粘膜表
面上皮(柱状细胞)与胃底腺上皮细胞排列紊乱。10 X40高
倍镜下脾虚模型组能见到一些胃粘膜细胞的损伤，尤对壁细
胞明显，如细胞皱缩，肪明莫出现缺损和胞质嗜酸性较正常组
降低，颜色由红色转为暗红色，甚至有些壁细胞呈现空泡化，
核浓密，而中药乐胃饮和香砂六君丸、西沙必利5组的壁细
胞损伤有不同程度的恢复，细胞增大，胞质嗜酸性降低。
结论“乐胃饮”具有使异常改变的胃肠激素Gas和枷t
的含量恢复至正常水平，并且对脾虚大鼠相关胃粘膜的病理
改变有促进其恢复作用。从而表明经验方“乐胃饮”作用机
理之一可能是通过对胃肠激素水平的影响从而能够达到有效
的治疗胃肠动力紊乱性疾病的目的。
Objective We use Chinese herb "Da huang" with bitter-cold property to creati the spleen-deficient model which is of Gastro-intestinal dysfunction in the light of west medicine, observing"Le Wei Yin'Y influence on serum Gastrin content and plasm Motilin content in rat and morphological chang in the gastral mucosal cell; investigating the relationship among'Te Wei Yin"-gut homrone-gastralintestinal dynamical and its protective function to gastral mucosa; explaining action mechanism of "Le Wei Yu"Twhich is treated gsatral-intestinal dynamical disorder disease.
Methods Seventy rats were divided at random into two big groups: normal group(ten) and experimental group (sixty). We feed experimental rats on 200% Da huang decotion After two weeks, we create sucesessfuly spleen-deficient model.Then We picked tats eyeballs to collect blood, measuring respectively Gastrin, Motilin, which are used as two indexes for our research on the mechanisum of treatment of "Le Wei Yin"at the level of gastral-intestinal hormone. After that, we kill the rats by means of decollation, pick out stomach tissu, put into 100ml L formalin promptly to fix tie tissue, then dehydration by alclhol, embed by paraffin, staining by Hematoxylin-esoin(HE), make sections in order to investigate morphological chang in gastral mucosal cell.
Results (1)General state: During the experiment, rats of normal group have none abnormal phenomenon in the light of its appetite, activity, stool and
skin What's more, they all put up little by little. By contrast, rats of model group are withered and yellow, arch its back, losing its weight, poor appetite, loose stool, avertion to cold after one week. What's worse, after half and a week, rats got annus proplapse, stiff stool or loose stool but remaining its formation. (2)Gastral-intestinal hormone: a. Serum Gastrin content: There is a remarkablely difference (P<0.01) between modle group and control group which note that spllen-deficient rats 's serum Gastrin content is significantly low; There is a remarkablely difference between high, moderat dosage of "Le Wei Yin" and model groupHigh dosage compared with model group(P<0.01), moderate dosage compared with model group(P<0.05).From the angle of average, high dosage group is by far closer to normal group, b. Plasm Motilin content There is a remarkablely difference (P<0.01) between modle group and control group which note that spllen-deficient rats 's plasm Motilin content is significantly highPatent Chinese herb "Xiang sha liu jun wan" group is significantly differente from model group(P<0.01). High, moderate dosage of "Le Wei Yin" group possess remarkabley difference compared with model group (P<0.05), while low dosage of "Le Wei Yin" is significantly differente from normal group,and is not significantly differente from model group. (3)Under the inverted microscope: Through 100 times power lens, we observed sections of each group and did not find inflammatory infiltration such as lymphocyte, plasmablast etc. In spleen-deficient model group induced by "Da huang", We find superficial epithelial cell and fundic gland epithelial cell of gastral mucosal cell arranging disorder. Through 400 times high power lens, We find some gastral mucosal cell damage, espetially
the parietal cell, while that in "Le Wei Yin", "Xiang sha liu jun", "Xi sha bi li" groups, cellular damage resume to some degree and some of them get large with cytoplast acidophilic decreasing.
Conclusions From above, experierre prescription''Le Wei Yin" possess the function of changing abnormal gut-hormon GAS and MOT content into normal state and the function of resuming gastral mucosal pathological change of spleen-deficient rats. So it mate clear that influence on gut-hormon may be one of action mechanism of "Le Wei Yin"efrective treatment of gut-intestinal dynamical disorder disease.
引文
1 孙维峰，王小春，韦嵩，等．中药升降汤对小白鼠肠组织胃动素及生长抑素的影响．广州中医药大学学报，1998，15(4)：270～272．
2 贾华群，蒋筱强，熊本京，等．肝胆通对胆结石豚鼠体内胃肠澈素水平的影响．中国中西医结合外科杂志，1999，5(1)：47～49．
3 赵根生．试论消化动力中药．陕西中医函授，1997，4(1)：18～19．
4 赵军宁，王建华，陈蔚文，等．调理脾胃方药作用的新视觉：对胃肠激素的影响．中药药理与临床，1996，12(1)：42～45．
5 张航向，任平，李源，等．中药对胃肠激素及胃肠运动的调节．World chin J．Digestol，2000，8(10)：1141～1144．
6 陈奇．中药药理现代研究方法学．第1版．北京：人民卫生出版社，1993．1103～1105．
7 胡素坤，等．中国中医研究院三十年论文文选．第1版．北京：中医古籍出版社，1985．352．
8 危北海．中医脾胃学说应用研究．第1版．北京：北京出版社，1993．98．
9 (英)C．F．A卡林著，孔庆雷译．组织病理学与组织化学技术手册．第1版．北京：科学出版社，1982．183～184．
10 许国铭，李石．现代消化病学．第1版．北京：人民军医出版社，1998．682，796，854．
11 钱可大．功能性消化不良研究进展．浙江中西医结合杂志，2000，11(10)：595～597．
12 许国铭，李石．现代消化病学．第1版．北京：人民军医出版社，1998．862．
13 王本祥．现代中药药理．第1版．天津：天津科学技术出版社，1999．1205．
14 王本祥．现代中药药理．第1版．天津：天津科学技术出舨社，1999．542．
15 李乾构．FD辨证论治探析．中国中西医结合脾胃杂志，1996，4(2)：115．
16 秦新裕，雷勇．胃肠肽类激素与胃肠动力．中国实用外科杂志，2001，21(6)：329～331．
17 金敬善，王广才，张绳祖，等．血清中胃泌素水平与脾虚证的关系．中西医结合杂志，1982，2(1)：25～26．
18 Liddle RA. Cholecystokinin Cell. Annu Rev Physical, 1997, 59:221～242.
19 ChfistotidesND.Motilm In:Guthonmones.2Edition London:Churchill Living estone, 1981. 273.
20 危北海．中医脾胃学说的应用研究．第1版．北京：北京出版社，1993．80．
21 冯五金，苏娟萍，李玉莲，等胃肠激素与中医证型关系的临床研究．中医杂志，1997，38(5)：298～299．
22 易崇勤，叶百宽，金敬善，等．四君子汤对脾虚大鼠粘膜酶组织化学及血浆胃肠激素的影响．北京中医药大学学报，1997，11(6)：31～34．
2 贾华群，蒋筱强，熊本京，等．肝胆通对胆结石豚鼠体内胃肠澈素水平的影响．中国中西医结合外科杂志，1999，5(1)：47～49．
3 赵根生．试论消化动力中药．陕西中医函授，1997，4(1)：18～19．
4 赵军宁，王建华，陈蔚文，等．调理脾胃方药作用的新视觉：对胃肠激素的影响．中药药理与临床，1996，12(1)：42～45．
5 张航向，任平，李源，等．中药对胃肠激素及胃肠运动的调节．World chin J．Digestol，2000，8(10)：1141～1144．
6 陈奇．中药药理现代研究方法学．第1版．北京：人民卫生出版社，1993．1103～1105．
7 胡素坤，等．中国中医研究院三十年论文文选．第1版．北京：中医古籍出版社，1985．352．
8 危北海．中医脾胃学说应用研究．第1版．北京：北京出版社，1993．98．
9 (英)C．F．A卡林著，孔庆雷译．组织病理学与组织化学技术手册．第1版．北京：科学出版社，1982．183～184．
10 许国铭，李石．现代消化病学．第1版．北京：人民军医出版社，1998．682，796，854．
11 钱可大．功能性消化不良研究进展．浙江中西医结合杂志，2000，11(10)：595～597．
12 许国铭，李石．现代消化病学．第1版．北京：人民军医出版社，1998．862．
13 王本祥．现代中药药理．第1版．天津：天津科学技术出版社，1999．1205．
14 王本祥．现代中药药理．第1版．天津：天津科学技术出舨社，1999．542．
15 李乾构．FD辨证论治探析．中国中西医结合脾胃杂志，1996，4(2)：115．
16 秦新裕，雷勇．胃肠肽类激素与胃肠动力．中国实用外科杂志，2001，21(6)：329～331．
17 金敬善，王广才，张绳祖，等．血清中胃泌素水平与脾虚证的关系．中西医结合杂志，1982，2(1)：25～26．
18 Liddle RA. Cholecystokinin Cell. Annu Rev Physical, 1997, 59:221～242.
19 ChfistotidesND.Motilm In:Guthonmones.2Edition London:Churchill Living estone, 1981. 273.
20 危北海．中医脾胃学说的应用研究．第1版．北京：北京出版社，1993．80．
21 冯五金，苏娟萍，李玉莲，等胃肠激素与中医证型关系的临床研究．中医杂志，1997，38(5)：298～299．
22 易崇勤，叶百宽，金敬善，等．四君子汤对脾虚大鼠粘膜酶组织化学及血浆胃肠激素的影响．北京中医药大学学报，1997，11(6)：31～34．