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[目的]探讨疏肝健脾方对腹泻型肠易激综合征(diarrhea-predominant pattern irritable bowel syndrome,D-IBS)大鼠的治疗作用及肥大细胞、5-HT通路的影响。[方法]1日龄SD雄性大鼠随机分为正常组、模型组、疏肝健脾方组。采用母婴分离联合束缚应激方法建立D-IBS大鼠模型。造模成功后,疏肝健脾方组给予疏肝健脾方灌胃(2.33g·100g~(-1)·d~(-1)),正常组和模型组则给予等体积的生理盐水灌胃。灌胃结束后,测定各组大鼠粪便含水量、直结肠球囊扩张刺激时腹壁撤退反射(abdominal withdrawal reflex,AWR)评分及腹外斜肌肌电(electromyography,EMG)积分变化率。并分别采用甲苯胺蓝染色检测结肠肥大细胞表达,酶联免疫法(ELISA)测定结肠5-HT水平,蛋白免疫印记法(Western blot,WB)检测结肠5-HT3R(5-hydroxytryptamine receptor 3,5-HT3R)、5-HT4R(5-hydroxytryptamine receptor 4,5-HT4R)蛋白表达。[结果]疏肝健脾方组大鼠粪便含水量较模型组降低(P<0.05);直结肠内球囊压力在40、60、80mmHg(1mmHg=0.133kPa)时,AWR评分及EMG积分变化率较模型组均降低(P<0.05);疏肝健脾方组结肠肥大细胞数目、5-HT含量、5-HT3AR及5-HT3BR蛋白表达较模型组均降低(P<0.05),5-HT4R蛋白表达较模型组升高(P<0.05)。[结论]疏肝健脾方对D-IBS大鼠有较好治疗作用,其作用机制可能与降低结肠肥大细胞数目、5-HT的含量、5-HT3AR及5-HT3BR的表达,升高5-HT4R的表达有关。
[Objective] To explore the therapeutic effect of Shuganjianpi Recipe on diarrhea-predominant pattern irritable bowel syndrome (D-IBS) in rats and its effect on mast cells and 5-HT pathway. [Method] One-day-old SD male rats were randomly divided into normal group, model group and Shugan Jianpi Fang group. D-IBS rat model was established by the method of separation of maternal-fetal separation and restraint stress. After successful modeling, Shugan Jianpi Fang group was given Shugan Jianpi Decoction (2.33g · 100g -1 · d -1), normal group and model group were given equal volume of normal saline Gavage. After the end of gavage, the water content of feces of rats in each group was measured. The score of abdominal withdrawal reflex (AWR) and the change of electromyography (EMG) score of the extracorporeal oblique muscle were measured. The colon mast cells were detected by toluidine blue staining, the colon 5-HT level was measured by enzyme-linked immunosorbent assay (ELISA) and the level of 5-HT3R (5-hydroxytryptamine receptor 3,5 -HT3R), 5-HT4R (5-hydroxytryptamine receptor 4,5-HT4R) protein expression. [Result] The water content of feces of Shuganjianpi Decoction group was lower than that of model group (P <0.05). The changes of AWR score and EMG score of 40,6,80mmHg (1mmHg = 0.133kPa) (P <0.05). The number of colon mast cells, the content of 5-HT, the expressions of 5-HT3AR and 5-HT3BR in Shuganjianpi Decoction group were lower than those in model group (P <0.05) HT4R protein expression than the model group increased (P <0.05). [Conclusion] Shuganjianpi Recipe has a good therapeutic effect on D-IBS rats, its mechanism may be related to reducing the number of colon mast cells, the content of 5-HT, the expression of 5-HT3AR and 5-HT3BR, increasing 5 -HT4R expression.