温脾健肾方对腹泻型肠易激综合征大鼠结肠黏膜5-羟色胺及其受体的影响

Effects of Wenshen Jianpi Decoction on 5-hydroxytryptamine and its receptors in colon mucosa of rats with diarrhea-predominant irritable bowel syndrome

目的观察温肾健脾方对腹泻型肠易激综合征(IBS-D)模型大鼠结肠黏膜5-羟色胺(5-HT)、5-羟色胺3受体(5-HT3R)、5-羟色胺4受体(5-HT4R)表达的影响。方法将40只SD雄性大鼠随机分为正常组、模型组、中药组及西药组,对模型组、中药组及西药组大鼠采用AL-Chaer ED造模方法联合番泻叶灌胃、夹尾刺激、束缚的综合方法进行IBS-D模型复制,造模成功后各组分别予0.9%Na Cl溶液、温肾健脾方、得舒特灌胃2周。观察大鼠一般情况,检测大鼠腹壁撤退反射(abdominal withdrawal reflex,AWR)评分及粪便含水量,并取大鼠结肠黏膜,采用酶联免疫法检测5-HT,免疫组化法检测5-HT3R、5-HT4R的表达。结果与正常组比较,模型组大鼠出现大便稀溏、毛色晦暗、食欲下降等变化,同时大鼠AWR评分升高、粪便含水量升高,结肠黏膜的5-HT水平升高、5-HT3R、5-HT_4R光密度值升高,差异均有统计学意义(P<0.05,P<0.01)。与模型组比较,中药组大鼠AWR评分、粪便含水量及结肠黏膜5-HT、5-HT3R、5-HT4R的表达降低,差异均有统计学意义(P<0.05,P<0.01)。结论温肾健脾方对IBS-D模型大鼠的治疗作用可能是通过影响5-HT及其受体表达,调节肠神经系统,减弱肠道运动及分泌,降低肠道内脏高敏感,从而达到综合治疗的作用。

Objective To observe the effects of Wenshen Jianpi Decoction on 5-hydroxytryptamine （5-HT） , 5-HT3 receptor （5-HT3R） , and 5-HT4 receptor （5- HT4R） in colon mucosa of rats with diarrhea-predominant irritable bowel syndrome （IBS-D）. Methods Forty male Sprague-Dawley rats were divided randomly into four groups as the normal group, model group, Chinese medicine group, and westem medicine group. Except the normal group, IBS-D model was copied in the other groups by an integrated approach which combined the AL-Chaer ED modeling method, intragastrie administration with senna soup, tail tamping stimulation and constraint stress. After successful modeling, the normal and model group were administrated with 0.9% NaCl, the Chinese medicine group was treated with Wenshen Jianpi Decoction, and the western medicine group was treated with pinaverium bromide, with a course of two weeks for intragastric administration. The general situation of rats was observed, the score of abdominal withdrawal reflex（AWR） and the fecal moisture content in rats were measured. The colonic mueosa of rats were collected, the expression of 5-HT was determined by enzymelinked immunosorbent assay and the expression of 5-HT3 R and 5-HT4 R were determined by immunohistochemieal assay. Results Compared with the normal group, the rats in the model group appeared loose stool, dull color of fur and appetite loss, meanwhile the AWR scores of rats and fecal moisture content increased, the levels of 5-HT and the optical density of 5-HT3R and 5-HT4R in the colonic mucosa increased, with statistically significant differences （ P 〈 0.05, P 〈 0.01 ）. Compared with the model group, the AWR scores, fecal moisture content and expression of 5-HT,5-HT3R,5-HT4R in the colonic mucosa of the Chinese medicine group decreased,with statistically significant differences （P 〈 0.05, P 〈 0.01 ）. Conclusion The comprehensive effects of Wenshen Jianpi Decoction in the treatment of model rats with IBS-D may involve with the regulating 5-HT and its receptors, regulating enteric nervous system, reducing bowel movement and secretion, and decreasing intestinal visceral hypersensitivity.