| 引用本文: |
罗容, 钟峰, 殷伶, 叶勇.基于全转录组测序分析电针大肠俞、天枢改善慢传输型便秘大鼠肠动力的作用机制[J].湖南中医药大学学报,2026,46(1):70-81[点击复制] |
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| 基于全转录组测序分析电针大肠俞、天枢改善慢传输型便秘大鼠肠动力的作用机制 |
| 罗容,钟峰,殷伶,叶勇 |
| (湖南中医药大学第一附属医院, 湖南 长沙 410007;湖南中医药大学第一中医临床学院, 湖南 长沙 410007) |
| 摘要: |
| 目的 基于全转录组测序技术，观察电针大肠俞、天枢对慢传输型便秘（STC）大鼠肠道动力的影响，并探讨其潜在的分子调控机制。方法 9只SD大鼠中随机抽取3只作为空白组，其余大鼠采用洛哌丁胺灌胃法构建STC大鼠模型，造模成功后再随机分为模型组和电针组，每组3只。电针组选取双侧大肠俞、天枢进行电针干预，连续14 d；空白组予以生理盐水灌胃，模型组予以盐酸洛哌丁胺混悬液灌胃维持模型稳定，电针组在模型组的基础上选取双侧大肠俞、天枢进行电针干预，均连续干预14 d。观察并记录大鼠首粒红便排出时间，干预14 d后24 h内粪便粒数及质量、小肠推进率；采集结肠组织进行全转录组测序，筛选差异表达基因；运用GO功能和KEGG通路富集分析探讨电针干预的生物学功能，筛选关键核心靶点并分析其调控关系，选取关键差异表达基因进行RT-qPCR验证。结果 与空白组比较，模型组、电针组首粒红便排出时间均延长（P＜0.05，P＜0.01），24 h粪便粒数及质量、小肠推进率降低（P＜0.05，P＜0.01），证实STC模型构建成功；与模型组比较，电针组24 h粪便粒数及质量、小肠推进率均升高（P＜0.05，P＜0.01）。全转录组测序筛选出412个差异mRNA、25个差异lncRNA、6 789个差异circRNA及12个差异miRNA。GO功能与KEGG通路富集分析表明，模型组的铁死亡与缺氧诱导因子-1（HIF-1）信号通路异常激活相关；电针的效应机制与Janus激酶-信号转导及转录激活因子（JAK-STAT）、核因子-κB（NF-κB）信号通路，以及补体与凝血级联反应的调控密切相关。RT-qPCR验证显示，与空白组比较，模型组激活转录因子3（Atf3）、丝氨酸肽酶抑制剂Kazal 1型（Spink1）、活化T细胞核因子1（Nfatc1）及微小RNA-27a-5A（miR-27a-5p）相对表达量显著上调（P＜0.01），同源盒基因A6（HoxA6）、蛋白激酶cAMP激活催化亚基β（Prkacb）下调（P＜0.05）；与模型组比较，电针组Atf3、Spink1、Nfatc1及miR-27a-5p相对表达量显著下调（P＜0.01），HoxA6、Prkacb上调（P＜0.05），趋势与测序结果一致。结论 电针大肠俞、天枢可能通过调控HIF-1、JAK-STAT及NF-κB等信号通路，并影响miR-27a-5p等关键基因表达，协同改善肠道炎症、能量代谢与神经可塑性，从而多靶点治疗STC大鼠肠道动力障碍。 |
| 关键词: 慢传输型便秘 电针 肠动力 全转录组测序 差异表达基因 |
| DOI：10.3969/j.issn.1674-070X.2026.01.009 |
| 投稿时间：2025-07-21 |
| 基金项目:国家自然科学基金面上项目（82374595）；湖南省自然科学基金项目（2024JJ6356）；湖南省卫生健康委员会卫生科研项目（W20243084）；湖南省卫生健康委员会高层次人才（湘卫函〔2024〕43号）。 |
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| Mechanism of action of electroacupuncture at "Dachangshu (BL25)" and "Tianshu (ST25)" in improving intestinal motility in rats with slow transit constipation based on whole transcriptome sequencing analysis |
| LUO Rong, ZHONG Feng, YIN Ling, YE Yong |
| (The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;The First Clinical College of Chinese Medicine, Hunan University of Chinese Medicine, Changsha, Hunan 410007, China) |
| Abstract: |
| Objective To observe the effects of electroacupuncture (EA) at "Dachangshu (BL25)" and "Tianshu (ST25)" on intestinal motility in rats with slow transit constipation (STC) based on whole transcriptome sequencing technology, and explore the underlying potential molecular regulatory mechanisms. Methods Three rats were randomly selected from nine SD rats as the blank group, while the remaining rats were used to establish STC rat model via loperamide gavage. After successful modeling, the model rats were randomly divided into the model group and the EA group, with three rats in each group. The EA group received intervention at bilateral "Dachangshu (BL25)" and "Tianshu (ST25)" for 14 consecutive days; the blank group received gavage of normal saline for 10 consecutive days; and the model group received gavage of loperamide hydrochloride suspension for 7 consecutive days. The time to the first red stool excretion was observed and recorded. After 14 days of intervention, the number and weight of feces within 24 hours, as well as the small intestinal transit rate, were measured. Colonic tissues were collected for whole transcriptome sequencing to screen differentially expressed genes (DEGs). Gene Ontology (GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed to explore the biological functions of EA intervention, to screen key core targets and analyze their regulatory relationships, and to select key DEGs for verification by quantitative reverse transcription polymerase chain reaction (RT-qPCR). Results Compared with the blank group, the time to the first red stool excretion was prolonged in both the model group and the EA group (P＜0.05, P＜0.01), while the number and weight of feces within 24 hours and the small intestine transit rate reduced (P＜0.05, P＜0.01), confirming the successful construction of the STC model. Compared with the model group, the number and weight within 24 hours, as well as the small intestine transit rate, increased in the EA group (P＜0.05, P＜0.01). Whole transcriptome sequencing identified 412 differentially expressed mRNAs, 25 differentially expressed lncRNAs, 6789 differentially expressed circRNAs, and 12 differentially expressed miRNAs. GO function and KEGG pathway enrichment analyses showed that ferroptosis in the model group was associated with abnormal activation of the hypoxia inducible factor-1 (HIF-1) signaling pathway; the effect mechanism of EA is closely related to the regulation of the Janus kinase-signal transducer and activator of transcription (JAK-STAT), nuclear factor-κB (NF-κB) signaling pathways, as well as the complement and coagulation cascade reactions. RT qPCR validation showed that compared with the blank group, the relative expression levels of activating transcription factor 3 (Atf3), serine peptidase inhibitor kazal type 1 (Spink1), nuclear factor of activated t cells 1 (Nfatc1), and microRNA-27a-5p (miR-27a-5p) in the model group were significantly upregulated (P＜0.01), while homeobox A6 (HoxA6) and protein kinase camp-activated catalytic subunit β (Prkacb) were downregulated (P＜0.05); Compared with the model group, the relative expression levels of Atf3, Spink1, Nfatc1, and miR-27a-5p in the electroacupuncture group were significantly downregulated (P＜0.01), while HoxA6 and Prkacb were upregulated (P＜0.05), consistent with the sequencing results. Conclusion Electroacupuncture at "Dachangshu (BL25)" and "Tianshu (ST25)" may regulate signaling pathways including HIF-1, JAK-STAT and NF-κB, and affect the expression of key genes such as miR-27a-5p, thereby synergistically ameliorating intestinal inflammation, energy metabolism and neural plasticity, and exerting a multi-target therapeutic effect on intestinal motility disorders in STC rats. |
| Key words: slow transit constipation electroacupuncture intestinal motility whole transcriptome sequencing differentially expressed genes |
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