摘要
目的探讨肠激安方对腹泻型肠易激综合征(IBS-D)大鼠结肠黏膜超微结构及上皮细胞闭锁小带蛋白(ZO-1)和紧密连接蛋白(CLDN1)表达的影响。方法将40只SPF级新生SD大鼠随机分为正常组、模型组、匹维溴铵组(18 mg·kg^(-1))和肠激安方高(33.48 g·kg^(-1))、低(16.74 g·kg^(-1))剂量组,共5组,每组8只,采用三因素(母婴分离+醋酸刺激+束缚)结合的方法,复制IBS-D大鼠模型。模型复制结束后,给予匹维溴铵和肠激安方连续灌胃14 d。用腹部回缩反射(AWR)评价内脏高敏状态,透射电镜(TEM)观察结肠黏膜超微结构的变化,采用免疫组化(IHC)、实时荧光定量聚合酶链式反应(RT-qPCR)法分析结肠组织ZO-1和CLDN1的表达。结果与模型组比较,匹维溴铵组、肠激安方高剂量组、肠激安方低剂量组大鼠AWR内脏敏感性评分降低(P<0.05,P<0.01);透射电镜观察结肠黏膜超微结构的结果显示,IBS-D模型组紧密连接不完整,部分有断裂,缝隙连接增宽,给予药物干预后,各组大鼠结肠黏膜中破坏的紧密连接结构得到一定程度的修复;与模型组比较,匹维溴铵组和肠激安方高、低剂量组结肠组织中ZO-1和CLDN1的mRNA相对含量升高(P<0.05,P<0.01),ZO-1、CLDN1的平均光密度(IOD)值升高(P<0.05,P<0.01)。结论肠激安方可改善IBS-D模型大鼠结肠黏膜超微结构异常,上调结肠黏膜上皮细胞ZO-1和CLDN1的表达。
Objective This study intends to investigate the effects of Changji’an formula on ultrastructure of colonic mucosa and levels of ZO-1 and CLDN1 in model rats with IBS-D.Methods 40 SPF grade SD rats were randomly divided into five groups,including the normal group,the model group,the pinaverium bromide group(18 mg·kg^(-1)),the high dose of Changji’an formula group(33.48 g·kg^(-1)),and the low dose of Changji’an formula group(16.74 g·kg^(-1)),with eight rats per group.The rat model of IBS-D was established by three-factors method:maternal separation,acetic acid stimulation and restraint stress.At the end of the modeling,all medication was given by gavage and lasted for 14 successive days After medication,AWR scores of rectal distention test were performed to evaluate the visceral sensitivity,the changes of ultrastructure of colonic mucosa were observed by TEM.Immunohistochemistry(IHC)was used to examine the expression of CLDN1 and ZO-1 in colonic tissues.The mRNA levels of CLDN1,ZO-1 in colon tissues were assessed using RT-qPCR.Results Compared with model group,AWR scores were decreased in pinaverium bromide group,high and low dose of Changji’an formula group(P<0.05,P<0.01).The TEM results showed that the tight junction of IBS-D model group appeared incomplete,disrupted and widen.Changji’an formula alleviated the impairment of ultrastructure of colonic mucosa in IBS-D model rats on a certain degree.Compared with model group,the mRNA levels of CLDN1 and ZO-1 in rats’colonic tissue were increased after administration of high and low dose of Changji’an formula and pinaverium bromide(P<0.05,P<0.01).The average optical density values of ZO-1 and CLDN1 with immunohistochemistry increased significantly(P<0.05,P<0.01).Conclusion Changji’an formula can improve the disorder ultrastructure of colonic mucosa and up-regulate levels of ZO-1 and CLDN1 in model rats with IBS-D.
作者
祝赫
柴玉娜
张毅靖
王永福
甘剑峰
周云松
唐洪梅
ZHU He;CHAI Yuna;ZHANG Yijing;WANG Yongfu;GAN Jianfeng;ZHOU Yunsong;TANG Hongmei(The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052 Henan,China;The First School of Clinical Medicine,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;The Second Affiliated Hospital of Guizhou University of Chinese Medicine,Guiyang 550000 Guizhou,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2021年第7期905-911,共7页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金面上项目(81673842,81703955)
广东省自然科学基金面上项目(2019A1515011745)。
