“补骨脂-肉豆蔻”醇提物调控CRF-CREB信号通路对肠易激综合征内脏高敏感大鼠发挥治疗作用的研究

A mechanism exploration of alcohol extracts of Psoralea corylifolia L and Nutmeg treating the irritable bowel syndrome visceral hypersensitivity rats based on CRF-CREB signaling pathway

目的:观察"补骨脂-肉豆蔻"醇提物对母子分离结合乙酸灌肠诱导肠易激综合征内脏高敏感大鼠CRF-CREB信号通路的作用。方法:将新生SD大鼠随机分为正常组、模型组、"补骨脂-肉豆蔻"醇提物(15 g/kg、10 g/kg、5 g/kg)组和匹维溴铵(0.05g/kg)组。除正常组外,其余各组采用母子分离结合乙酸灌肠法复制内脏高敏感大鼠模型,于造模后第6周起,给药组分别灌胃给予相应的受试药物,正常组和模型组给予等量溶媒,1天1次,连续2周。采用直肠扩张法评价其腹部收缩反射(AWR)阈值,HE染色观察大鼠结肠组织病理学变化;免疫荧光检测大鼠结肠促肾上腺皮质激素释放因子(CRF),CRF 1型受体(CRFR1)蛋白的表达;RT-qP CR技术检测蛋白激酶A(PKA)、环磷腺苷效应元件结合蛋白(CREB)、c-fos mRNA的表达;Western blot技术检测PKA、CREB、p-CREB、c-fos蛋白的表达。结果:"补骨脂-肉豆蔻"醇提物(15 g/kg、10 g/kg)不仅能降低结肠组织中CRF、CRFR1蛋白的表达,还能降低结肠组织中PKA、CREB、p-CREB、c-fos mRNA和蛋白表达。结论:"补骨脂-肉豆蔻"对肠易激综合征内脏高敏感性大鼠有一定的治疗作用,其机制与调节结肠组织中CRF-CREB信号通路有关。

Objective： To study the effect of alcohol extracts from Psoralea corylifolia L ﹠ Nutmeg on the corticotropin-releasing factor（ CRF） and c AMP response element binding protein（ CREB） signal transduction pathway in rats with the visceral hypersensitivity. Methods： Newborn SD rats were randomly divided into six groups,including the control group,the model group,alcohol extracts from Psoralea corylifolia L ﹠Nutmeg（ 15 g/kg,10 g/kg,5 g/kg）,and 0. 05 g/kg pinaverium bromide. Except the control group,rats in other groups received the separation and acetic acid to copy the visceral hypersensitivity models. At 6 th week after model rats were made,corresponding drugs were given to rats by intragastric administration once a day for two weeks. Then the threshold of abdominal withdrawal reflex was measured at 6 th week by the rectal distension. Pathological changes were observed by Hematein Eosin（ HE）; expressions of CRF,CFRR1 were determined by immunofluorescence technique; Moreover,real-time fluorescent quantitative PCR was performed to detect expressions of proteine kinase A（ PKA）,CREB,and c-fos mRNA,and Western blotting was used to determine expressions of PKA,CREB,p-CREB,and c-fos protein. Results： Compared with the model group,alcohol extracts from Psoralea corylifolia L ﹠ Nutmeg（ 15 g/kg,10 g/kg） not only effectively decreased expressions of CRF,CRFR1,but also inhibited expressions of PKA,CREB,p-CREB,c-fos in the colon. Conclusion： Alcohol extracts from Psoralea corylifolia L ﹠ Nutmeg has certain effect on the visceral hypersensitivity rats,which might be related to decreasing expressions of CRF,CRFR1 and p-CREB in the colon and blocking CRF-CREB signal transduction pathway.