基于转录组测序技术探讨电针对应激性胃溃疡模型大鼠的分子机制

Molecular Mechanism of Stress-induced Gastric Ulcers Involved in Electroacupuncture Based on Transcriptome Sequencing

目的探讨电针针刺对应激性胃溃疡大鼠模型胃黏膜的保护作用及转录组学分析分子机制。方法选取SPF级成年雄性Wistar大鼠,随机分为模型组(n=8)、空白组(n=8)、电针组(n=8)。电针组予以电针针刺内关和足三里治疗,每日30 min。空白组常规喂养,无任何处置。模型组同期进行异氟烷麻醉干预。采用束缚-水浸应激法在干预结束后建立动物模型。取胃组织并测量溃疡面积,计算溃疡指数,HE染色法观察胃黏膜形态学改变,采用转录组测序技术检测各组大鼠胃组织基因表达,筛选差异表达基因,并进行基因GO功能分类以及KEGG代谢通路富集分析。结果在预电针针刺后,胃溃疡面积明显低于模型组。高通量测序结果的GO富集分析表明胃溃疡的产生和预电针对胃溃疡的治疗过程中,涉及生物过程、细胞组分和分子功能中的多个方面,同时也表明胃溃疡的形成与电针对其的治疗是多组分、多基因相互作用的结果。KEGG富集结果表明,PI3K-Akt和MAPK等信号通路在胃溃疡的发生和治疗过程中起着重要作用。结论本研究初步探讨了胃溃疡形成及预电针针刺缓解胃溃疡大鼠的分子调控机制,为进一步解析电针对胃溃疡治疗的分子机制研究奠定了良好的基础。

Objective:To investigate the protective effect of electroacupuncture on gastric mucosa in rats with stress gastric ulcer and the molecular mechanism of transcriptome analysis.Methods:SPF adult male Wister rats were randomly divided into model group(n=8),blank group(n=8)and electroacupuncture group(n=8).Elec⁃troacupuncture group was treated with electroacupuncture on“Neiguan”(PC 6)and“Zusanli”(ST 36)for 30 min/d.Blank group:routine feeding without any treatment.The model group received isoflurane anesthesia interven⁃tion at the same time.The animal model was established after the intervention by shackle-water stress method.Gastric tissue was taken and ulcer area was measured,ulcer index was calculated,morphological changes of gastric mucosa were observed by HE staining,gene expression in gastric tissue of each group was detected by transcrip⁃tome sequencing technology,differentially expressed genes were screened,and gene GO functional classification and KEGG metabolic pathway enrichment analysis were performed.Results:After pre-electroacupuncture,gas⁃tric ulcer area was significantly lower than that of the model group.The GO enrichment analysis of high-through⁃put sequencing results showed that the generation of gastric ulcer and the treatment of gastric ulcer by electrothera⁃py involved many aspects of biological processes,cellular components and molecular functions,and that the forma⁃tion of gastric ulcer and the treatment of gastric ulcer by electrotherapy were the result of multi-component and multi-gene interaction.KEGG enrichment results indicated that pI3K-Akt and MAPK signaling pathways played an important role in the occurrence and treatment of gastric ulcer.Conclusion:This study preliminarily discusses the molecular regulation mechanism of gastric ulcer formation and alleviating gastric ulcer by pre-elec⁃troacupuncture in rats,which has laid a good foundation for further analyzing the molecular mechanism of elec⁃troacupuncture for gastric ulcer treatment.