摘要
目的观察微小核糖核酸(Micro-ribonucleic acid,miRNA)在腹泻型肠易激综合征(Irritable bowel syndrome-diarrhea,IBS-D)大鼠和健脾合剂治疗后的大鼠结肠组织中表达的差异,预测健脾合剂治疗IBS-D过程中参与的miRNA及对应的基因,为寻找IBS-D的潜在治疗靶点提供基础。方法将大鼠随机分为空白组、模型组和中药组,每组8只。模型组与中药组均采用急慢性刺激法+番泻叶灌胃制造IBS-D模型,中药组于造模后予以健脾合剂灌胃,连续14天;观察并记录大鼠一般情况、体重及粪便Bristol积分;采用苏木精-伊红(Hematoxylin-eosin staining,HE)染色法观察各组大鼠结肠组织病理学变化;对各组大鼠结肠组织进行转录组测序,筛选差异表达的miRNA并进行映射;对筛选出的共同miRNA对应的靶基因进行GO(Gene oncology)功能富集分析和KEGG(Kyoto Encyclopedia of Genes and Genomes)通路富集分析。结果与模型组相比,中药组大鼠体重显著增加(P<0.01),粪便Bristol积分明显下降(P<0.01),结肠组织病理损伤减轻。与空白组相比,模型组差异表达的基因共109个,其中80个上调,29个下调。与模型组相比,中药组差异表达的基因共109个,其中19个上调,90个下调。对两次比较的基因进行映射发现,模型组中有74个miRNA升高而7个降低,这些miRNA的变化均在中药干预后得到逆转。对靶基因进行GO功能富集分析显示健脾合剂作用于大鼠后主要参与突触囊泡定位与转运,胞质运输、生长性发育负调控、糖运输等过程。KEGG通路富集分析显示丝裂原活化蛋白激酶(Mitogen-activated protein kinase,MAPK)、Wnt、Hippo、肿瘤坏死因子(Tumor necrosis factor,TNF)、氧毒素、环磷酸腺苷(cyclic AMP,cAMP)等通路被富集。结论健脾合剂干预治疗后,IBS-D相关的miRNA表达谱发生了显著的变化;健脾合剂可能通过调控MAPK信号通路、Wnt信号通路等在治疗IBS-D过程中发挥保护作用。筛选核心基因和预测与之相互作用的miRNA为IBS-D发病机制和健脾合剂治疗靶点的研究提供了理论基础。
Objective To observe the difference in the expression of micro-ribonucleic acid(miRNA)in the colon tissue of irritable bowel syndrome-diarrhea(IBS-D)rats and rats treated with Jianpi Mixture,and predict The miRNA and corresponding genes involved in the treatment of IBS-D with Jianpi Mixture,providing a basis for finding potential therapeutic targets for IBS-D.Methods The rats were randomly divided into a blank group,a model group and a traditional Chinese medicine group,with 8 rats in each group.Both the model group and the Chinese medicine group used acute and chronic stimulation+senna leaf gavage to make the IBS-D model.The Chinese medicine group was gavaged with Jianpi Mixture after model building for 14 days.General condition,body weight and Bristol score were observed and recorded.Hematoxylin-eosin(HE)staining was used to observe the histopathological changes of colon in each group of rats.Transcriptome sequencing of rat colon tissue in each group to screen and map differentially expressed miRNAs.Perform GO(Gene oncology)functional enrichment analysis and KEGG(Kyoto Encyclopedia of Genes and Genomes)pathway enrichment analysis on the target genes corresponding to the screened common miRNAs.Results Compared with the model group,the weight of the rats in the Chinese medicine group increased significantly(P<0.01),and the Bristol score of the feces decreased significantly(P<0.01).The pathological damage of the colon tissue of rats in the Chinese medicine group was significantly reduced.Compared with the blank group,there were 109 differentially expressed genes in the model group,of which 80 were up-regulated and 29 were down-regulated.Compared with the model group,there were 109 differentially expressed genes in the traditional Chinese medicine group,of which 19 were up-regulated and 90 were down-regulated.Mapping the genes of the two comparisons found that 74 miRNAs in the model group were increased and 7 were decreased.The changes of these miRNAs were all reversed after the intervention of traditional Chinese medicine.The GO function enrichment analysis of target genes showed that Jianpi Mixture was mainly involved in synaptic vesicle positioning and transport,cytoplasmic transport,negative regulation of growth and development,and sugar transport after acting on rats.KEGG pathway enrichment analysis showed that MAPK,Wnt,Hippo,TNF,oxygen toxin,cAMP and other pathways were enriched.Conclusion After the intervention of Jianpi Mixture,IBS-D-related miRNA expression profiles have changed significantly;Jianpi Mixture may play a protective role in the treatment of IBS-D by regulating the MAPK signaling pathway and Wnt signaling pathway.Screening the core genes and predicting the miRNAs interacting with them provide a theoretical basis for the study of the pathogenesis of IBS-D and the therapeutic targets of Jianpi Mixture.
作者
王萌
吴婷婷
丁姮月
梁国强
杨欣
张培培
孙宏文
Wang Meng;Wu Tingting;Ding Hengyue;Liang Guoqiang;Yang Xin;Zhang Peipei;Sun Hongwen(Suzhou TCM Hospital Affiliated to Nanjing University of Chinese Medicine,Suzhou 215009,China;the Affiliated Suzhou Science&Technology Town Hospital of Nanjing Medical University,Suzhou 215153,China;Suzhou Academy of Wumen Chinese Medicine,Suzhou 215009,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2023年第6期2071-2080,共10页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
苏州市科技局、苏州市卫生局面上项目(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
苏州市科技局面上项目(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。
