养荣润肠舒对便秘大鼠肠道c-Kit蛋白含量影响实验研究

Experimental Study on Effect of Yangrong Runchangshu Decoction in Constipation Mice Intestinal c-Kit Protein Content

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：Using Western blot to detect the expression of c-Kit protein in the colon of mice,and explore the mechanism of traditional Chinese medicine compound Yangrong Runchangshu Decoction in treatment of slow transit constipation. Method：Select 24 mice were random Ly divided into：normal group,model group and Yangrong Runchangshu Decoction group,8 rats in each group,half male and half female,The model group and Yangrong Runchangshu Decoction group mice were treated by method of morphine hydrochloride subcutaneous injection molding. As the 10 days observation period,The experimental mice last 2 hours after ad ministration,were sacrificed,the abdo minal cavity was opened immediately,separation of large intestine,the mice proximal colon tissue of about 2 cm,physiological saline,frozen in liquid nitrogen and stored at-70 ℃,standby. Changes with the technique and method of expression of westernblot protein was detected by c-Kit in the mouse colon,and the application of chemi imager 5500 gel image analysis software（America）analysis,record each protein electrophoresis bands of gray value,quantitative analysis.Results：the expression of c-Kit protein in model group in significantly decreased compared with normal group（P0.01）,c-Kit protein expression Yangrong Runchangshu Decoction treatment group increased significantly after,is about 2.93 times than the model group（P0.01）,but still significantly lower than the normal group,the difference was significant（P0.05）.Conclusion：Yangrong Runchangshu Decoction can regulate c-Kit protein expression to increase the number and function of ICC,and enhance colonic peristalsis,to the treatment of slow transit constipation effect.