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目的:应用蛋白免疫印迹法检测大鼠结肠c-Kit蛋白表达的变化,探讨中药复方养荣润肠舒治疗慢传输型便秘的作用机制。方法:选取24只大鼠随机分为:正常组、模型组和养荣润肠舒组,每组8只,雌雄各半,其中模型组和养荣润肠舒组大鼠均采用盐酸吗啡皮下注射的方法造模。观察期为10 d,实验大鼠末次给药后2 h,脱颈处死,立即打开腹腔,分离大小肠,选取大鼠近端结肠组织约2 cm,生理盐水冲洗后,置于液氮中冷冻,-70℃保存备用。采用western-blot的技术方法检测大鼠结肠c-Kit蛋白表达的变化情况,并应用Chemi Imager 5500凝胶成像分析软件(America)分析,记录每条蛋白电泳带的灰度值,进行定量分析。结果:c-Kit蛋白在模型组中的表达较正常组明显降低(P<0.01),治疗后养荣润肠舒组c-Kit蛋白的表达显著升高,约为模型组的2.93倍(P<0.01),但仍明显低于正常组,两者差异显著(P<0.05)。结论:养荣润肠舒能通过调节c-Kit蛋白表达来增加ICC的数量和功能,进而增强结肠蠕动,达到治疗慢传输型便秘的作用。
OBJECTIVE: To detect the changes of c-Kit protein expression in rat colon by Western blotting and to explore the mechanism of traditional Chinese medicine Yirong Runchangshu on slow transit constipation. Methods: Twenty-four rats were randomly divided into normal group, model group and Yangrun Runchangshu group, with 8 rats in each group, male and female rats in each group. Rats in model group and Yunrong Runchang Shu group were treated with morphine hydrochloride Injection method modeling. The observation period was 10 days. The experimental rats were sacrificed 2 hours after the last administration, the abdominal cavity was opened and the small intestine was separated. The proximal colon tissue of rats was selected to be about 2 cm in length. After washing with physiological saline, the rats were frozen in liquid nitrogen , -70 ℃ save spare. Western-blot technique was used to detect the expression of c-Kit protein in rat colon. The gray value of each protein electrophoretic band was recorded by Chemi Imager 5500 gel imaging analysis software (America), and quantitative analysis was performed. Results: The expression of c-Kit protein in model group was significantly lower than that in normal group (P <0.01). After treatment, the expression of c-Kit protein in Yangrun Runchang Shu group was significantly increased, about 2.93 times that of model group <0.01), but still significantly lower than the normal group, the difference was significant (P <0.05). Conclusion: Yangrun Runchang Shu can increase the number and function of ICC by regulating the expression of c-Kit protein, and then enhance colon peristalsis to achieve the effect of slow-transit constipation.