Regulatory mechanism of electroacupuncture on colonic function and CGRP-RAMP1 signaling pathway in diarrhea-predominant irritable bowel syndrome rats with liver-depression and spleen-deficiency

doi:10.16098/j.issn.0529-1356.2026.06.016

PDF(15807 KB)

Acta Anatomica Sinica ›› 2026, Vol. 57 ›› Issue (5) : 639-647. DOI: 10.16098/j.issn.0529-1356.2026.06.016

Histology and embryology and Developmental Biology

Regulatory mechanism of electroacupuncture on colonic function and CGRP-RAMP1 signaling pathway in diarrhea-predominant irritable bowel syndrome rats with liver-depression and spleen-deficiency

WANG Qian-ru¹, GUO Huan-qin¹, HAO Li-fang^1*, ZHOU Yu-zheng²

Author information +

History +

Abstract

Objective To evaluate how electroacupuncture (EA) mitigates visceral hypersensitivity in a rat model of diarrhea-predominant irritable bowel syndrome (IBS-D) characterized by liver-depression and spleen-deficiency, and to elucidate its molecular mechanism in targeting calcitonin gene-related peptide (CGRP), calcitonin receptor-like receptor (CRLR), and receptor activity-modifyingprotein 1 (RAMP1) receptor complex axis. Methods Forty SPF male SD rats were randomly divided into the blank control group (n=10) and the model establishment group (n=30). The IBS-D model was established using a comprehensive stress method. Successfully modeled rats were randomly subdivided into the model group (n=9), electroacupuncture (EA) group (n=9), and Western medicine(pinaveriumbromide)(WM) group (n=8). Body weight, fecal water content, and glass bead expulsion time were evaluated. Abdominal withdrawal reflex (AWR) scores and the corresponding area under the curve (AUC) were used to evaluate visceral hyperalgesia. Meanwhile, colonic morphological alterations were examined by HE staining. Furthermore, the transcriptional and translational profiles of CGRP, CRLR, and RAMP1 were analyzed utilizing Real-time PCR, Western blotting, and immunohistochemical assays with 6 independent biological samples from each group (n=6). Results Compared with the blank group, the model establishment group exhibited significantly decreased body weight, alongside significantly increased fecal water content, AWR score, and shortened glass bead expulsion time (all P<0.01). HE staining showed mild edema and inflammatory infiltration in the colonic mucosa, with significantly increased histological scores (P<0.01). A profound upregulation was observed in both the transcript and protein levels of colonic CGRP, CRLR, and RAMP1 (P<0.01). In contrast to the untreated model rats, both EA and pinaverium bromide treatments markedly alleviated the aforementioned behavioral deficits and colonic structural damages (P<0.05 or P<0.01), and significantly down-regulated the protein and mRNA expression levels of CGRP, CRLR, and RAMP1 (P<0.01). Regarding overall efficacy, the WM group showed superior improvement in fecal water content compared to the EA group (P<0.05), while the EA group exhibited stronger analgesic efficacy in alleviating severe visceral hypersensitivity (80 mmHg) (P<0.05). Conclusion Electroacupuncture successfully ameliorates intestinal dysmotility and blunts visceral hyperalgesia in IBS-D rats. Mechanistically, this protective effect is closely tied to the suppression of the hyperactive CGRP/CRLR/RAMP1 signaling cascade within the colonic tissue.

Key words

Diarrhea-predominant irritable bowel syndrome

/ Electroacupuncture / Calcitonin gene-related peptide / Receptor activity-modifying protein 1 / Comprehensive stress method / Western blotting / Rat

Cite this article

EndNote

Ris (Procite)

Bibtex

Download Citations

WANG Qian-ru, GUO Huan-qin, HAO Li-fang, ZHOU Yu-zheng. Regulatory mechanism of electroacupuncture on colonic function and CGRP-RAMP1 signaling pathway in diarrhea-predominant irritable bowel syndrome rats with liver-depression and spleen-deficiency[J]. Acta Anatomica Sinica. 2026, 57(5): 639-647 https://doi.org/10.16098/j.issn.0529-1356.2026.06.016

References

［1］Farooq A, Naveed M, Chowdhury R, et al. Enhancing inflammatory bowel disease (IBD) competency in gastroenterology fellowship through traditional, digital, and experiential learning pathways ［J］. Inflamm Bowel Dis, 2026, 32(3): izag043.

［2］Jia L, Cao S, Chen L, et al. Electroacupuncture inhibits the NLRP3/caspase-1/ASC signaling pathway in a mouse model of inflammatory bowel disease ［J］. Acupunct Med, 2026, 44(2): 9645284261423205.

［3］Hu XF, Zhang H, Yu LL, et al. Electroacupuncture reduces anxiety associated with inflammatory bowel disease by acting on cannabinoid CB1 receptors in the ventral hippocampus in mice ［J］. Front Pharmacol, 2022, 13: 919553.

［4］Deng N, Xie S, Liu Q, et al. The intestinal microbiota modulates the visceral sensitivity involved in IBS induced by restraint combined with tail clustering ［J］. Front Cell Infect Microbiol, 2025, 15: 1549617.

［5］Qu S, Yu Z, Zhou Y, et al. Gut microbiota modulates neurotransmitter and gut-brain signaling ［J］. Microbiol Res, 2024, 287: 127858.

［6］Edvinsson L, Holm A. Understanding RAMP1’s role in hormonal migraine may improve CGRP-targeted therapies ［J］. Expert Rev Clin Pharmacol, 2026, 19(2): 97-99.

［7］Liu X, Chen J, Zou L, et al. Acupuncture accelerates wound healing via CGRP-RAMP1-TSP1-mediated macrophage M2 polarization ［J］. Chin Med, 2025, 20(1): 192.

［8］Fattori V, Zaninelli TH, Rasquel-Oliveira FS, et al. Nociceptor-to-macrophage communication through CGRP/RAMP1 signaling drives endometriosis-associated pain and lesion growth in mice ［J］. Sci Transl Med, 2024, 16(772): eadk8230.

［9］Li ZhX, He XM, Chen JF, et al. Fuzi Lizhong Pills modulate NF-κB/Notch1/COX-2 pathway to improve diarrhea-predominant irritable bowel syndrome in rats ［J］. Journal of Guangzhou University of Traditional Chinese Medicine, 2026, 43(3): 715-722. (in Chinese)

李志雄, 何晓铭, 陈锦锋, 等. 附子理中丸调控NF-κB/Notch1/COX-2通路改善大鼠腹泻型肠易激综合征［J］. 广州中医药大学学报, 2026, 43(3): 715-722.

［10］Zhu YX, Yin SY, Gao TY, et al. Research progress of animal models of ulcerative colitis ［J］. Chinese Journal of Gastroenterology and Hepatology, 2026, 35(3): 322-330. (in Chinese)

朱元兴, 尹思语, 高天佑, 等. 溃疡性结肠炎动物模型研究进展［J］. 胃肠病学和肝病学杂志, 2026, 35(3): 322-330.［11］China Association of Acupuncture-Moxibustion. Name and location of acupoints commonly used in laboratory animals-Part 2: Rats ［J］. Acupuncture Research, 2021, 46(4): 351-352. (in Chinese)

中国针灸学会. 实验动物常用穴位名称与定位第2部分: 大鼠［J］. 针刺研究, 2021, 46(4): 351-352.

［12］Xu ShY, Bian RL, Chen X. Methodology of Pharmacological Experiments ［M］. Beijing: People’s Medical Publishing House, 2002. (in Chinese)

徐叔云, 卞如濂, 陈修. 药理实验方法学［M］. 北京:人民卫生出版社, 2002.

［13］Leonard F, Srinivasan S, Liu X, et al. Design and in vitro characterization of multistage silicon-PLGA budesonide particles for inflammatory bowel disease ［J］. Eur J Pharm Biopharm, 2020, 151: 61-72.

［14］Chen L, Yan Y, Shi X, et al. Central amygdala HDAC6 contributes to visceral hypersensitivity and affective comorbidities in IBS-like rats ［J］. Int Immunopharmacol, 2026, 176: 116466.

［15］Najjar SA, Kildegaard H, Talati A, et al. Enteric and sympathetic nervous system pathways mediate early life stress effects on gut motility and pain: Mechanistic findings with human correlation ［J］. Gastroenterology, 2026, 170(4): S0016-5085(26)00205-2.

［16］Chen T. Clinical efficacy of self-made Shugan Lipi Decoction on ulcerative colitis of liver stagnation and spleen deficiency type based on brain-gut axis theory ［D］. Nanchang:Jiangxi University of Chinese Medicine, 2023. (in Chinese)

陈婷. 基于脑肠轴理论探讨自拟疏肝理脾汤对肝郁脾虚型溃疡性结肠炎的临床疗效［D］. 南昌：江西中医药大学, 2023.

［17］He JS, Zhou JM, Yang YT, et al. Pathogenesis and traditional Chinese medicine treatment of inflammatory bowel disease complicated with depression ［J］. Pharmacy Today, 2026: 1-9. (in Chinese)

何佳穗, 周家梅, 杨雅婷, 等. 炎症性肠病合并抑郁症发病机制与中医药治疗研究进展［J］. 今日药学, 2026: 1-9.

［18］Yang XY, Fu ShP, Wu JY, et al. Effect of electroacupuncture at "Tianshu" combined with semaglutide on up-regulating pancreatic GLP-1R expression and lowering blood glucose in type 2 diabetic rats ［J］. Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology, 2026: 1-18. (in Chinese)

杨心仪, 傅淑平, 吴嘉悦, 等. 电针“天枢”联合司美格鲁肽上调胰腺GLP-1R表达降低2型糖尿病大鼠血糖的研究［J］. 世界科学技术-中医药现代化, 2026: 1-18.

［19］Verma A, Inslicht SS, Bhargava A. Gut-brain axis: Role of microbiome, metabolomics, hormones, and stress in mental health disorders ［J］. Cells, 2024, 13(17): 1436.

［20］Qin Y, Chen X, Yu Z, et al. Spinal RAMP1-mediated neuropathic pain sensitisation in the aged mice through the modulation of CGRP-CRLR pain signalling ［J］. Heliyon, 2024, 10(16): e35862.

［21］Suekane A, Saito Y, Nakahata S, et al. CGRP-CRLR/RAMP1 signal is important for stress-induced hematopoiesis ［J］. Sci Rep, 2019, 9(1): 429.

［22］Zhang SJ, Geng MM, Li QW, et al. Oxidative stress impairs the expansion of regulatory T cells in active vitiligo via dysregulated CGRP-RAMP1-Gαi3 signaling ［J］. Free Radic Biol Med, 2026, 245: 18-29.

［23］Wang XM, Li Y, Zhang DL, et al. Electroacupuncture regulates enteric nervous system and tight junction proteins in functional dyspepsia rats ［J］. Acta Anatomica Sinica, 2024, 55(2): 184-191. (in Chinese)

王小明, 李毅, 张大力, 等. 电针对功能性消化不良大鼠肠神经系统及紧密连接蛋白的调节作用［J］. 解剖学报, 2024, 55(2): 184-191.

［24］Zhang Y, Liu JH, Chen T, et al. Expression and significance of CGRP in spinal cord dorsal horn of rats with visceral hypersensitivity ［J］. Acta Anatomica Sinica, 2025, 56(1): 45-52. （in Chinese）

张莹, 刘建华, 陈婷, 等. 内脏高敏感大鼠脊髓背角CGRP的表达及意义［J］. 解剖学报, 2025, 56(1): 45-52.