天津医药 ›› 2020, Vol. 48 ›› Issue (12): 1169-1174.doi: 10.11958/20201095

• 实验研究 • 上一篇    下一篇

CGRP、AchE在功能性排便障碍大鼠模型中的表达

李雪1,2,3,耿学斯1,3,程一乘1,2,4,刘薇1,2,刘立阳1,2,刘仍海1,2△   

  1. 1北京中医药大学(邮编100029);2北京中医药大学东方医院肛肠科;3北京中医药大学厦门医院肛肠科;4中日友好医院
  • 收稿日期:2020-04-26 修回日期:2020-08-27 出版日期:2020-12-15 发布日期:2020-12-13
  • 通讯作者: 刘仍海 E-mail:liurenghai@163.com
  • 作者简介:李雪(1992),女,博士在读,主要从事中医药防治大肠肛门疾病研究
  • 基金资助:
    国家自然科学基金面上项目(81273757)

The expressions of CGRP and AchE in a rat model of functional defecation disorder

LI Xue1, 2, 3, GENG Xue-si1, 3, CHENG Yi-cheng1, 2, 4, LIU Wei1, 2, LIU Li-yang1, 2, LIU Reng-hai1, 2△   

  1. 1 Beijing University of Chinese Medicine, Beijing 100029, China; 2 Department of Anorectal Surgery, Dongfang Hospital, Beijing University of Chinese Medicine; 3 Department of Anorectal, Xiamen Hospital, Beijing University of 
    Chinese Medicine; 4 China-Japan Friendship Hospital
  • Received:2020-04-26 Revised:2020-08-27 Published:2020-12-15 Online:2020-12-13
  • Contact: LIU Reng-hai E-mail:liurenghai@163.com

摘要: 目的 观察降钙素基因相关肽(CGRP)和胆碱酯酶(AchE)在功能性排便障碍(FDD)大鼠模型结直肠中的表达,探讨FDD发生的可能机制。方法 SD大鼠随机分为空白对照组、低纤维饮食组、利多卡因组、模型组,空白对照组予以普通饲料饲养,其余组采用低纤维饲料饲养;利多卡因组及模型组于饲养第63天,分别予以2 mL 2%利多卡因、0.1%亚甲蓝注射液肛周及直肠周围间隙注射1次。观察大鼠粪便性状、质量及排便功能;酶联免疫吸附试验(ELISA)检测结、直肠中AchE含量;实时荧光定量PCR(qPCR)检测结、直肠中CGRP mRNA的表达,免疫组化及Western blot法检测CGRP蛋白的表达。结果 肛周注射后,与空白对照组相比,低纤维饮食组和利多卡因组的粪便干结、呈深褐色,模型组的粪便干硬、呈黑褐色,粪便质量均下降。模型组模拟球囊排出时间较其余组长,肛管直肠静息压较其余组低;结肠及直肠中AchE含量较空白对照组和低纤维饮食组低(P<0.05);结肠中CGRP mRNA表达水平高于空白对照组,与低纤维饮食组和利多卡因组差异无统计学意义;直肠中CGRP mRNA表达水平较其余组高;免疫组化检测CGRP蛋白表达高于其余组(P<0.05);而Western blot检测各组CGRP蛋白表达差异均无统计学意义。结论 大鼠结直肠中CGRP及AchE的表达异常可能与FDD发生的神经机制有关。

关键词: 降钙素基因相关肽, 乙酰胆碱酯酶, 结肠, 直肠, 功能性排便障碍

Abstract: Objective To observe the expressions of calcitonin gene-related peptide (CGRP) and cholinesterase (AchE) in colon and rectum of the rat model of functional defecation induced by low fiber diet combined with local methylene blue anal injection, and explore the possible mechanism of the occurrence of functional defecation disorder. Methods SD rats were randomly divided into blank control group, low-fiber diet group, lidocaine group and model group. The blank control group was fed with ordinary feed, and the other groups were fed with low-fiber feed. The lidocaine group and model group were given 2 mL of 2% lidocaine and 0.1% methylene blue injection once around the perianal and perirectal space respectively on the 63rd day of rearing. Fecal characteristics, fecal quality and defecation function of rats were observed. ELISA was used to detect AchE content in colon and rectum of rats. Real-time PCR (qPCR) was used to detect the expressions of CGRP mRNA and protein in rat colon and rectum respectively. Immunohistochemistry and Western blot assay were used to evaluate CGRP protein expression. Results After perianal injection, compared with the blank control group, the stools were dry and hard, and dark brown in the low-fiber diet group and lidocaine group, and the quality of the stool was decreased in the model group. The simulated balloon discharge was longer in the model group than that of the other groups, and the resting pressure in colon and rectum was lower than that of the other groups. The AchE contents in the colon and rectum were lower in model group than those of the blank control group and low-fiber diet group (P<0.05). The expression of CGRP mRNA in the rectum was increased than that of the blank control group, but there was no significant difference between the blank control group, low-fiber diet group and lidocaine group. The expression of CGRP mRNA in rectum was higher than that of the other groups. The expression of CGRP protein detected by immunohistochemistry was higher in model group than that of the other groups (P<0.05), while the expression of CGRP protein detected by Western blot assay was not statistically significant between four groups. Conclusion The abnormal expressions of CGRP and AchE in rat colon and rectum may be related to the neural mechanism of functional defecation disorder.

Key words: calcitonin gene-related peptide, acetylcholinesterase, colon, rectum, functional defecation disorder

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