普芦卡必利治疗小鼠慢传输型便秘的机制研究？

Study on the Mechanism of Prucalopride for the Treatment of Slow Transit Constipation in Mice

目的研究普芦卡必利对慢传输型便秘（slow transit constipation,SLC）小鼠结肠 Caj al 间质细胞结构、数量的影响,探讨普芦卡必利治疗 SLC新机制。方法注射盐酸吗啡腹腔制备 SLC 小鼠模型,普芦卡必利灌胃治疗。观测治疗后小鼠排便；剪取近端结肠,HE 染色检测结肠 Cajal 间质细胞（ICC）结构变化, Real-time PCR法检测 c-kit表达,试剂盒法检测钙离子浓度变化。结果与正常组相比,模型组小鼠不排便动物数,8 h排便总数、20 h排便总数增多（P＜ 0.05）。普芦卡必利治疗后8 h排便总数、20 h排便总数较模型组都表现为升高（P ＜ 0.05）。HE染色结果显示模型组小鼠结肠 ICC出现基膜溶解,数量减少,其与周围细胞之间的紧密连接也被破坏,细胞核有不同程度萎缩；普芦卡必利治疗后,ICC 细胞数目和结构都有所恢复。c-kit表达检测显示,模型组 c-kit表达显著地性的降低（P ＜ 0.01）,普芦卡必利治疗后,c-kit 表达升高（P＜ 0.05）。钙离子浓度检测显示模型组钙离子浓度显著性的降低（P＜ 0.01）,普芦卡必利治疗后,钙离子浓度升高（P＜ 0.05）。结论普芦卡必利通过调节钙离子浓度保护ICC功能可能是其发挥对SLC治疗作用的机制之一。

Obj ective]To investigate the effect of prucalopride on the structure and quantity of interstitial cells of Cajal（ICC）in colon of slow transit constipation（SLC）in mice,and to explore its new mechanisms.[Methods]The mouse models of SLC were prepared by intraperitoneal inj ection of morphine hydrochloride. The mice with SLC received prucalopride by lavage.After the treatment,defecation of mice was observed. Proximal colon was clipped to detect the change of ICC structure by HE staining.Real-time PCR method was used to detect the expression of c-kit.Kit method was used to measure the change of Ca2＋ concentration.[Results]Compared with normal group,the number of mice without defecation,the total of 8h and 20h defe-cation in model group were increased（P〈 0.05）.Compared with model group,the total of 8h and 20h defeca-tion after prucalopride treatment were increased （P 〈 0.05）.HE staining showed that the ICC of colon in model group had the dissolving of basement membrane,and the number was reduced,and close connection be-tween ICC and the surrounding cells was destroyed,and nuclei of ICC had different degrees of atrophy.After prucalopride treatment,the number and structure of ICC were recovered.C-kit expression assay showed that the expression of c-kit in model group was significantly reduced（P〈 0.01）,and then increased after prucalo-pride treatment（P〈 0.05）.Ca2＋ concentration assay showed that Ca2＋ concentration in model group was sig-nificantly decreased（P〈 0.01）,and then increased after prucalopride treatment（P 〈 0.05）.[Conclusion]Regulating Ca2＋ concentration and protecting ICC function may be one of the mechanisms of prucalopride for the treatment of SLC.