摘要
目的基于miRNA-219a-5p、miRNA-338-3p探讨健脾合剂干预腹泻型肠易激综合征(Diarrhea-predominant irritable bowel syndrome,IBS-D)大鼠的作用机制。方法采用急-慢应激法联合番泻叶灌胃法制备IBS-D大鼠模型,分组后给予健脾合剂干预。实验过程中及治疗完成后,记录大鼠一般情况、体质量及粪便性状;采用直肠扩张下腹壁回缩反射(Abdominal withdrawal reflex,AWR)评分检测大鼠的疼痛阈值,评估IBS-D大鼠的内脏敏感性;采用显色基质鲎试剂检测大鼠血液中的细菌内毒素含量,观察其肠道通透性变化;观察大鼠结肠黏膜组织病理,对其肠黏膜状态进行评估;并使用RT-PCR法检测大鼠结肠黏膜miR-219a-5p、miR-338-3p的含量。结果粪便性状评分、内脏敏感性测定、组织病理结果表明IBS-D大鼠模型制备成功。与空白组比较,模型组大鼠大便稀溏甚则水样,体质量下降;健脾合剂治疗后,低剂量组及高剂量组大鼠大便成形,粪便Bristol评分显著降低,体质量有所增加。内脏敏感性结果显示,IBS-D大鼠内脏疼痛阈值下降;健脾合剂干预后,低剂量组及高剂量组大鼠内脏疼痛阈值有所升高,内脏敏感性降低。显色基质鲎试剂检测结果显示,IBS-D大鼠血清内毒素含量升高;健脾合剂干预后,低剂量组及高剂量组大鼠内毒素含量较前下降,肠道通透性降低。大鼠结肠粘膜病理结果表明,IBS-D大鼠结肠组织黏膜上皮轻度破损,局部可见水肿。健脾合剂治疗后,低剂量组及高剂量组大鼠结肠黏膜上述情况得到有效改善。Realtime PCR法检测大鼠结肠粘膜结果显示,IBS-D大鼠结肠黏膜miR-219a-5p、miR-338-3p水平显著降低,健脾合剂治疗后,高剂量组大鼠miR-219a-5p、miR-338-3p含量有所升高。结论健脾合剂对改善IBS-D症状具有一定的作用,健脾合剂能够增加IBS-D大鼠结肠黏膜miR-219a-5p、miR-338-3p水平,提示其可能通过对miR-219a-5p、miR-338-3p的调控降低IBS-D大鼠的肠道通透性及内脏敏感性,从而发挥疗效。
Objective To explore the mechanism of Jianpi Mixture in interfering with diarrhea-predominant irritable bowel syndrome(IBS-D)model rats based on miRNA-219a-5p and miRNA-338-3p.Methods The IBS-D rat model was prepared by the acute-slow stress method combined with senna gavage method.After the rats were divided into groups,they were given the intervention of Jianpi Mixture.During the experiment and after the treatment,the general condition,body weight and fecal traits of the rats were recorded.In this experiment,the pain threshold of rat viscera was detected by abdominal withdrawal reflex(AWR),and the visceral sensitivity of IBS-D rats was evaluated.The chromogenic matrix limulus reagent was used to detect the bacterial endotoxin content in the blood of rats,and the changes in intestinal permeability were observed.The pathology of the colon mucosa of the rats was observed,and the state of the intestinal mucosa was evaluated.Finally,RT-PCR was used to detect the contents of miR-219a-5p and miR-338-3p in the colon mucosa of rats.Results The results of fecal trait score,visceral sensitivity measurement,and HE staining of colon tissue indicated that the IBS-D rat model was successfully prepared.Compared with the blank group,the stools of the rats in the IBS-D model group became watery and their body weight decreased.After treatment with Jianpi Mixture,the stools of the rats in the low-dose and high-dose groups were formed,their stool bristol scores were significantly reduced,and their body weights also increased.Visceral sensitivity results showed that the visceral pain threshold of IBS-D rats decreased;after the intervention of Jianpi Mixture,the visceral pain threshold of rats in the low-dose and high-dose groups increased,and the visceral sensitivity decreased.The test results of the chromogenic matrix limulus reagent showed that the serum endotoxin content of IBS-D rats increased;after the intervention of Jianpi Mixture,the endotoxin content of the low-dose group and the high-dose group decreased,and the intestinal permeability decreased.The pathological results of the rat colonic mucosa showed that the mucosal epithelium of the colon tissue of IBS-D rats was slightly damaged,with local edema and atypical hyperplasia of local epithelial cells.After the treatment of Jianpi Mixture,the above-mentioned conditions of the colonic mucosa of rats in the low-dose group and the highdose group were effectively improved.Real-time PCR detection of rat colonic mucosa showed that the contents of miR-219a-5p and miR-338-3p in colonic mucosa of IBS-D rats were significantly reduced.After treatment with Jianpi Mixture,the contents of miR-219a-5p and miR-338-3p in the high-dose group increased.Conclusion Jianpi Mixture has an effect on improving the symptoms of IBS-D.It can increase the content of miR-219a-5p and miR-338-3p in the colonic mucosa of IBS-D rats.It is suggested that Jianpi Mixture may reduce the intestinal permeability and visceral sensitivity of IBS-D rats by regulating miR-219a-5p and miR-338-3p,thereby exerting curative effect.
作者
丁姮月
王萌
杨欣
梁国强
吴婷婷
张培培
孙宏文
Ding Hengyue;Wang Meng;Yang Xin;Liang Guoqiang;Wu Tingting;Zhang Peipei;Sun Hongwen(Suzhou TCM Hospital Affiliated to Nanjing University of Chinese Medicine,Suzhou 215009,China;Graduate School,Nanjing University of Chinese Medicine,Nanjing 210023,China;The Affiliated Suzhou Science&Technology Town Hospital of Nanjing Medical University,Suzhou 215153,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2023年第3期1102-1110,共9页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
苏州市科学技术局、市卫生局面上项目(KJXW2021045):探讨健脾合剂调控miRNA-219a-5p、miRNA-338-3p干预IBS-D大鼠的作用机制,负责人:丁姮月
苏州市科学技术局面上项目(SYS201777):脾虚型IBS-D肠道微生物的变化及健脾合剂的干预研究,负责人:孙宏文
国家中医药管理局面上项目([2019]62):吴门医派杂病流派传承工作室项目,负责人:葛惠男
