Journal of Traditional Chinese Medicine ›› 2026, Vol. 46 ›› Issue (4): 894-906.DOI: 10.19852/j.cnki.jtcm.2026.04.010
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Electroacupuncture alleviates intestinal inflammation in rats with ulcerative colitis by regulating colonic cell autophagy and gut microbiota
XU Huichao, HAO Jianheng, CHANG Boya, WANG Danni, LI Rangqian, CAO Yuxia, WANG Haijun(
), JI Laixi()
the Second Clinical College ,Shanxi University of Chinese Medicine Jinzhong 030619, China
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Received:2025-06-17Accepted:2025-11-18Online:2026-08-15Published:2026-08-08 -
Contact:Prof. JI Laixi, Prof. WANG Haijun, the Second Clinical College, Shanxi University of Chinese Medicine, Jinzhong 030619, China. jlx@sxtcm.edu.cn; whjdavid@163.com,Telephone: +86-18635855529
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Supported by:National Natural Science Foundation of China: Study on the Regulatory Mechanism of Cellular Autophagy of Acupoint Prescription for Intestinal Diseases in Regulating Intestinal Flora Dysbiosis in Ulcerative Colitis(82074549)
Cite this article
XU Huichao, HAO Jianheng, CHANG Boya, WANG Danni, LI Rangqian, CAO Yuxia, WANG Haijun, JI Laixi. Electroacupuncture alleviates intestinal inflammation in rats with ulcerative colitis by regulating colonic cell autophagy and gut microbiota[J]. Journal of Traditional Chinese Medicine, 2026, 46(4): 894-906.
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Figure 1 EA intervention improved colonic injury in UC rats A: comparison of DAI scores; B: comparison of colon length; C: comparison of colon histopathology assessed by HE staining. Green arrows indicate crypt structural damage, whereas yellow arrows indicate inflammatory cell infiltration; C1: Con group; C2: UC group; C3: EA group; C4: RAPA group; C5: 3-MA group; D: comparison of inflammatory cytokine levels; D1: TNF-α; D2: IL-β; D3: IL-2; D4: IL-10. Con group: rats were allowed free access to distilled water without UC induction and received intraperitoneal injections of an equal volume of 0.9% saline; UC group: Rats were allowed free access to a 5% DSS solution for 7 consecutive days to establish the UC model. After successful model establishment, they received intraperitoneal injections of an equal volume of 0.9% saline once daily for 3 consecutive days; EA group: after UC model establishment, rats received electroacupuncture at Zhongwan (CV12), bilateral Tianshu (ST25), and bilateral Shangjuxu (ST37), using alternating sparse-dense waves at 10/50 Hz for 20 min once daily for 3 consecutive days. They also received intraperitoneal injections of an equal volume of 0.9% saline; RAPA group: after UC model establishment, rats received intraperitoneal injections of RAPA at 2 mg/kg once daily for 3 consecutive days; 3-MA group: after UC model establishment, rats received intraperitoneal injections of 3-MA at 1.5 mg/100 g once daily for 3 consecutive days. DAI: disease activity index; HE: hematoxylin and eosin; TNF-α: tumor necrosis factor-α; IL-1β: interleukin-1β; DSS: dextran sulfate sodium; UC: ulcerative colitis; EA: electroacupuncture; RAPA: rapamycin; 3-MA: 3-methyladenine. The results were analyzed using one-way analysis of variance. Data are presented as mean ± standard deviation (n = 8). Compared with the Con group, aP < 0.01; compared with the UC group, bP < 0.01, cP < 0.05.
Figure 1 EA intervention improved colonic injury in UC rats A: comparison of DAI scores; B: comparison of colon length; C: comparison of colon histopathology assessed by HE staining. Green arrows indicate crypt structural damage, whereas yellow arrows indicate inflammatory cell infiltration; C1: Con group; C2: UC group; C3: EA group; C4: RAPA group; C5: 3-MA group; D: comparison of inflammatory cytokine levels; D1: TNF-α; D2: IL-β; D3: IL-2; D4: IL-10. Con group: rats were allowed free access to distilled water without UC induction and received intraperitoneal injections of an equal volume of 0.9% saline; UC group: Rats were allowed free access to a 5% DSS solution for 7 consecutive days to establish the UC model. After successful model establishment, they received intraperitoneal injections of an equal volume of 0.9% saline once daily for 3 consecutive days; EA group: after UC model establishment, rats received electroacupuncture at Zhongwan (CV12), bilateral Tianshu (ST25), and bilateral Shangjuxu (ST37), using alternating sparse-dense waves at 10/50 Hz for 20 min once daily for 3 consecutive days. They also received intraperitoneal injections of an equal volume of 0.9% saline; RAPA group: after UC model establishment, rats received intraperitoneal injections of RAPA at 2 mg/kg once daily for 3 consecutive days; 3-MA group: after UC model establishment, rats received intraperitoneal injections of 3-MA at 1.5 mg/100 g once daily for 3 consecutive days. DAI: disease activity index; HE: hematoxylin and eosin; TNF-α: tumor necrosis factor-α; IL-1β: interleukin-1β; DSS: dextran sulfate sodium; UC: ulcerative colitis; EA: electroacupuncture; RAPA: rapamycin; 3-MA: 3-methyladenine. The results were analyzed using one-way analysis of variance. Data are presented as mean ± standard deviation (n = 8). Compared with the Con group, aP < 0.01; compared with the UC group, bP < 0.01, cP < 0.05.
Figure 2 EA intervention promotes autophagy levels in the colons of UC rats A: representative transmission electron microscopy images showing the ultrastructure of colon tissues. Yellow arrows indicate autolysosomes; A1-A5: TEM images of colon tissues; A6-A10: magnified views of the corresponding regions in A1-A5; A1, A6: Con group; A2, A7: UC group; A3, A8: EA group; A4, A9: RAPA group; A5, A10: 3-MA group; B: comparison of ATG5, ATG12, LC3-II/LC3-I, and p62 protein expression levels; B1: Western blot bands of each sample; B2: relative expression level of ATG5 protein; B3: relative expression level of ATG12 protein; B4: relative expression level of LC3 protein; B5: relative expression level of p62 protein; C: comparison of ATG5, ATG12, LC3, and p62 mRNA expression levels; C1: relative expression level of ATG5 Mrna; C2: relative expression level of ATG12 mRNA; C3: relative expression level of LC3 mRNA; C4: relative expression level of p62 mRNA; D: comparison of PI3K, p-AKT/AKT, and p-mTOR/mTOR protein expression levels; D1: Western blot bands of each sample; D2: relative expression level of PI3K protein; D3: relative expression level of AKT protein; D4: relative expression level of mTOR protein. Con group: rats were allowed free access to distilled water without UC induction and received intraperitoneal injections of an equal volume of 0.9% saline; UC group: Rats were allowed free access to a 5% DSS solution for 7 consecutive days to establish the UC model. After successful model establishment, they received intraperitoneal injections of an equal volume of 0.9% saline once daily for 3 consecutive days; EA group: after UC model establishment, rats received electroacupuncture at Zhongwan (CV12), bilateral Tianshu (ST25), and bilateral Shangjuxu (ST37), using alternating sparse-dense waves at 10/50 Hz for 20 min once daily for 3 consecutive days. They also received intraperitoneal injections of an equal volume of 0.9% saline; RAPA group: after UC model establishment, rats received intraperitoneal injections of RAPA at 2 mg/kg once daily for 3 consecutive days; 3-MA group: after UC model establishment, rats received intraperitoneal injections of 3-MA at 1.5 mg/100 g once daily for 3 consecutive days. TEM: transmission electron microscopy; ATG5: autophagy related 5; ATG12: autophagy related 12; LC3-Ⅰ: microtubule-associated protein 1 light chain 3-Ⅰ; LC3-Ⅱ: microtubule-associated protein 1 light chain 3-Ⅱ; p62: sequestosome 1; PI3K: phosphatidylinositol 3-kinase; AKT: protein kinase B; p-AKT: phosphorylated protein kinase B; mTOR: mechanistic target of rapamycin; p-mTOR: phosphorylated mechanistic target of rapamycin; DSS: dextran sulfate sodium; UC: ulcerative colitis; EA: electroacupuncture; RAPA: rapamycin; 3-MA: 3-methyladenine. The results were analyzed using one-way analysis of variance. Data are presented as mean ± standard deviation (n = 8). Compared with the Con group, aP < 0.05, cP < 0.01; compared with the UC group, bP < 0.05, dP < 0.01.
Figure 2 EA intervention promotes autophagy levels in the colons of UC rats A: representative transmission electron microscopy images showing the ultrastructure of colon tissues. Yellow arrows indicate autolysosomes; A1-A5: TEM images of colon tissues; A6-A10: magnified views of the corresponding regions in A1-A5; A1, A6: Con group; A2, A7: UC group; A3, A8: EA group; A4, A9: RAPA group; A5, A10: 3-MA group; B: comparison of ATG5, ATG12, LC3-II/LC3-I, and p62 protein expression levels; B1: Western blot bands of each sample; B2: relative expression level of ATG5 protein; B3: relative expression level of ATG12 protein; B4: relative expression level of LC3 protein; B5: relative expression level of p62 protein; C: comparison of ATG5, ATG12, LC3, and p62 mRNA expression levels; C1: relative expression level of ATG5 Mrna; C2: relative expression level of ATG12 mRNA; C3: relative expression level of LC3 mRNA; C4: relative expression level of p62 mRNA; D: comparison of PI3K, p-AKT/AKT, and p-mTOR/mTOR protein expression levels; D1: Western blot bands of each sample; D2: relative expression level of PI3K protein; D3: relative expression level of AKT protein; D4: relative expression level of mTOR protein. Con group: rats were allowed free access to distilled water without UC induction and received intraperitoneal injections of an equal volume of 0.9% saline; UC group: Rats were allowed free access to a 5% DSS solution for 7 consecutive days to establish the UC model. After successful model establishment, they received intraperitoneal injections of an equal volume of 0.9% saline once daily for 3 consecutive days; EA group: after UC model establishment, rats received electroacupuncture at Zhongwan (CV12), bilateral Tianshu (ST25), and bilateral Shangjuxu (ST37), using alternating sparse-dense waves at 10/50 Hz for 20 min once daily for 3 consecutive days. They also received intraperitoneal injections of an equal volume of 0.9% saline; RAPA group: after UC model establishment, rats received intraperitoneal injections of RAPA at 2 mg/kg once daily for 3 consecutive days; 3-MA group: after UC model establishment, rats received intraperitoneal injections of 3-MA at 1.5 mg/100 g once daily for 3 consecutive days. TEM: transmission electron microscopy; ATG5: autophagy related 5; ATG12: autophagy related 12; LC3-Ⅰ: microtubule-associated protein 1 light chain 3-Ⅰ; LC3-Ⅱ: microtubule-associated protein 1 light chain 3-Ⅱ; p62: sequestosome 1; PI3K: phosphatidylinositol 3-kinase; AKT: protein kinase B; p-AKT: phosphorylated protein kinase B; mTOR: mechanistic target of rapamycin; p-mTOR: phosphorylated mechanistic target of rapamycin; DSS: dextran sulfate sodium; UC: ulcerative colitis; EA: electroacupuncture; RAPA: rapamycin; 3-MA: 3-methyladenine. The results were analyzed using one-way analysis of variance. Data are presented as mean ± standard deviation (n = 8). Compared with the Con group, aP < 0.05, cP < 0.01; compared with the UC group, bP < 0.05, dP < 0.01.
Figure 3 EA intervention regulated the structure and composition of gut microbiota in UC rats A: comparison of rank abundance curves; B: comparison of Alpha diversity. B1 ACE; B2: Chao1; B3: Simpson; C: comparison of Beta diversity; C1: PCA; C2: PCoA; D: comparison of differences in the gut microbiota; D1: phylum levels; D2: genus levels; E: Spearman correlation analysis. Con group: rats were allowed free access to distilled water without UC induction and received intraperitoneal injections of an equal volume of 0.9% saline; UC group: Rats were allowed free access to a 5% DSS solution for 7 consecutive days to establish the UC model. After successful model establishment, they received intraperitoneal injections of an equal volume of 0.9% saline once daily for 3 consecutive days; EA group: after UC model establishment, rats received electroacupuncture at Zhongwan (CV12), bilateral Tianshu (ST25), and bilateral Shangjuxu (ST37), using alternating sparse-dense waves at 10/50 Hz for 20 min once daily for 3 consecutive days. They also received intraperitoneal injections of an equal volume of 0.9% saline; RAPA group: after UC model establishment, rats received intraperitoneal injections of RAPA at 2 mg/kg once daily for 3 consecutive days; 3-MA group: after UC model establishment, rats received intraperitoneal injections of 3-MA at 1.5 mg/100 g once daily for 3 consecutive days. ACE: abundance-based coverage estimator; Chao1: Chao1 richness estimator; PCA: principal component analysis; PCoA: principal coordinates analysis. The results were analyzed using one-way analysis of variance and Spearman correlation analysis. Data are presented as mean ± standard deviation (n = 8). Compared with the Con group, aP < 0.01; compared with the UC group, bP < 0.01.
Figure 3 EA intervention regulated the structure and composition of gut microbiota in UC rats A: comparison of rank abundance curves; B: comparison of Alpha diversity. B1 ACE; B2: Chao1; B3: Simpson; C: comparison of Beta diversity; C1: PCA; C2: PCoA; D: comparison of differences in the gut microbiota; D1: phylum levels; D2: genus levels; E: Spearman correlation analysis. Con group: rats were allowed free access to distilled water without UC induction and received intraperitoneal injections of an equal volume of 0.9% saline; UC group: Rats were allowed free access to a 5% DSS solution for 7 consecutive days to establish the UC model. After successful model establishment, they received intraperitoneal injections of an equal volume of 0.9% saline once daily for 3 consecutive days; EA group: after UC model establishment, rats received electroacupuncture at Zhongwan (CV12), bilateral Tianshu (ST25), and bilateral Shangjuxu (ST37), using alternating sparse-dense waves at 10/50 Hz for 20 min once daily for 3 consecutive days. They also received intraperitoneal injections of an equal volume of 0.9% saline; RAPA group: after UC model establishment, rats received intraperitoneal injections of RAPA at 2 mg/kg once daily for 3 consecutive days; 3-MA group: after UC model establishment, rats received intraperitoneal injections of 3-MA at 1.5 mg/100 g once daily for 3 consecutive days. ACE: abundance-based coverage estimator; Chao1: Chao1 richness estimator; PCA: principal component analysis; PCoA: principal coordinates analysis. The results were analyzed using one-way analysis of variance and Spearman correlation analysis. Data are presented as mean ± standard deviation (n = 8). Compared with the Con group, aP < 0.01; compared with the UC group, bP < 0.01.
Figure 4 FMT improved colonic injury in UC rats A: comparison of DAI scores; B: comparison of colon length; C: comparison of colon histopathology assessed by HE staining. Green arrows indicate crypt structural damage, and yellow arrows denote inflammatory cell infiltration. C1: FMT-EA group; C2: FMT-UC group; D: comparison of inflammatory cytokine levels; D1: TNF-α; D2: IL-β; D3: IL-2; D4: IL-10; E: comparison of protein expression levels of ATG5, ATG12, LC3-II/LC3-I and p62; E1: Western blot bands of each sample; E2: ATG5 protein; E3: ATG12 protein; E4: LC3 protein; E5: p62 protein; F: comparison of protein expression levels of PI3K, p-AKT/AKT and p-mTOR/mTOR; F1: Western blot bands of each sample; F2: PI3K protein; F3: m-TOR protein; F4: AKT protein. FMT-UC group: UC rats received fecal microbiota from UC donor rats by oral gavage for 7 consecutive days; FMT-EA group: UC rats received fecal microbiota from EA-treated donor rats by oral gavage for 7 consecutive days. DAI: disease activity index; HE: hematoxylin and eosin; FMT: fecal microbiota transplantation; UC: ulcerative colitis; EA: electroacupuncture; ATG5: autophagy-related protein 5; ATG12: autophagy-related protein 12; LC3: microtubule-associated protein 1 light chain 3; p62: sequestosome 1; PI3K: phosphoinositide 3-kinase; AKT: protein kinase B; p-AKT: phosphorylated AKT; mTOR: mechanistic target of rapamycin; p-mTOR: phosphorylated mTOR. Differences between two groups were analyzed using a Student t test. Data are presented as mean ± standard deviation (n = 8). Compared with the FMT-UC group, aP < 0.01, bP < 0.05.
Figure 4 FMT improved colonic injury in UC rats A: comparison of DAI scores; B: comparison of colon length; C: comparison of colon histopathology assessed by HE staining. Green arrows indicate crypt structural damage, and yellow arrows denote inflammatory cell infiltration. C1: FMT-EA group; C2: FMT-UC group; D: comparison of inflammatory cytokine levels; D1: TNF-α; D2: IL-β; D3: IL-2; D4: IL-10; E: comparison of protein expression levels of ATG5, ATG12, LC3-II/LC3-I and p62; E1: Western blot bands of each sample; E2: ATG5 protein; E3: ATG12 protein; E4: LC3 protein; E5: p62 protein; F: comparison of protein expression levels of PI3K, p-AKT/AKT and p-mTOR/mTOR; F1: Western blot bands of each sample; F2: PI3K protein; F3: m-TOR protein; F4: AKT protein. FMT-UC group: UC rats received fecal microbiota from UC donor rats by oral gavage for 7 consecutive days; FMT-EA group: UC rats received fecal microbiota from EA-treated donor rats by oral gavage for 7 consecutive days. DAI: disease activity index; HE: hematoxylin and eosin; FMT: fecal microbiota transplantation; UC: ulcerative colitis; EA: electroacupuncture; ATG5: autophagy-related protein 5; ATG12: autophagy-related protein 12; LC3: microtubule-associated protein 1 light chain 3; p62: sequestosome 1; PI3K: phosphoinositide 3-kinase; AKT: protein kinase B; p-AKT: phosphorylated AKT; mTOR: mechanistic target of rapamycin; p-mTOR: phosphorylated mTOR. Differences between two groups were analyzed using a Student t test. Data are presented as mean ± standard deviation (n = 8). Compared with the FMT-UC group, aP < 0.01, bP < 0.05.
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