摘要
目的基于miRNA-29a探讨肠激安方对腹泻型肠易激综合征(diarrhea-predominant irritable bowel syndrome,IBS-D)模型大鼠的干预作用。方法采用三因素(母婴分离、醋酸刺激、束缚应激)法复制IBS-D大鼠模型,分组后给予肠激安方干预,给药结束后定时考察大鼠的体重、糖水偏好值、自主活动行为特征,并通过腹部回缩反射(abdominal withdrawal reflex,AWR)直肠扩张实验评估IBS-D大鼠的内脏敏感性。酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)法测定大鼠血浆中二胺氧化酶(diamine oxidase,DAO)和D-乳酸(D-lactate,D-LA)的含量;RT-qPCR法检测结肠中miRNA-29a的含量。结果AWR评分、结肠组织HE染色结果表明三因素法复制大鼠IBS-D模型成功。与空白组比较,IBS-D模型组大鼠的体重明显降低,糖水偏好值显著降低,内脏敏感性评分明显升高;与模型组比较,匹维溴铵组、肠激安方高剂量组、肠激安方低剂量组的大鼠体重升高,糖水偏好值升高,AWR内脏敏感性评分降低。HE染色结果显示,各组大鼠结肠黏膜上皮细胞组织形态学无明显的病理变化。ELISA结果显示,与空白组比较,模型组血浆中D-LA和DAO的含量升高;与模型组比较,匹维溴铵组、肠激安方高剂量组血浆中D-LA明显降低,匹维溴铵组、肠激安方高剂量组、肠激安方低剂量组血浆中DAO含量明显降低;与空白组比较,模型组结肠中miRNA-29a的含量增加;与模型组比较,匹维溴铵组、肠激安高剂量组、肠激安低剂量组大鼠的结肠组织中miRNA-29a的含量降低。结论肠激安方对改善IBS-D脾虚泄泻具有一定的作用,肠激安方能够降低IBS-D大鼠结肠组织中miRNA-29a的含量,提示肠激安方干预IBS-D的作用机制可能与其对miRNA-29a的调控有关。
Objective To explore the intervention effects of Changji’an formula on model rats with diarrhea predominant irritable bowel syndrome(IBS-D)based on miRNA-29 a.Methods The rat model with IBS-D was established by using three-factors method,including maternal separation,acetic acid stimulation and restraint stress.After grouping,Changji’an formula was given for intervention.After administration,body weight,sucrose preference and autonomous activity behavior of rats were measure regularly.The visceral sensitivity of IBS-D rats was evaluated with rectal dilatation experiment of abdominal withdrawal reflex(AWR).The contents of diamine oxidase(DAO)and D-lactate(D-LA)in serum of rats was detected by ELISA.The content of miRNA-29 a in colon tissues was detected by using RT-qPCR.Results The AWR score and HE staining results of colon tissues showed that the three-factors method was successful in replicating the rat model with IBS-D.Compared with blank group,the body weight of IBS-D rats in the model group was obviously reduced,the value of sucrose preference was significantly reduced,and the value of visceral sensitivity increased obviously.Compared with the model group,the body weight and the value of sucrose preference increased in both high and low dose of Changji’an formula group,as well as the pinaverium bromide group,while the AWR scores decreased in each group.The results of HE staining in colon tissues showed that the histomorphology of colonic mucosal epithelial cells of rats in each group showed no obvious pathological changes.The results of ELISA showed that:Compared with control group,the plasma contents of DAO and D-LA increased in the model group;Compared with model group,the plasma contents of D-LA in piveronium bromide group and high dose of Changji’an formula group obviously reduced,the plasma contents of DAO obviously decreased in piveronium bromide group and high,low dose of Changji’an formula group;Compared with blank group,the content of miRNA-29 a of colon tissues in model group increased;Compared with model group,the contents of miRNA-29 a of colon tissues decreased in piveronium bromide group and high,low dose of Changji’an formula group.Conclusion Changji’an formula has some certain effects on IBS-D rats with diarrhea due to spleen deficiency,it can reduced the content of miRNA-29 a of colon tissues in IBS-D rats.These results suggest that the mechanism of action of Changji’an formula on IBS-D rats may be related to its regulation of miRNA-29 a.
作者
祝赫
石玉莹
吴映秀
肖茜
唐洪梅
ZHU He;SHI Yuying;WU Yingxiu;XIAO Xi;TANG Hongmei(Department of Pharmarcy,The first Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China)
出处
《环球中医药》
CAS
2020年第8期1309-1315,共7页
Global Traditional Chinese Medicine
基金
国家自然科学基金面上项目(81673842)
广东省自然科学基金面上项目(2019A1515011745)。
关键词
腹泻型肠易激综合征
肠激安方
小分子核糖核酸29a
腹部回缩反射
二胺氧化酶
Diarrhea-predominant irritable bowel syndrome
Changji'an formulaMicro ribonucleic acid-29a
Abdominal withdrawal reflex
Diamine oxidase
