Journal of Traditional Chinese Medicine ›› 2025, Vol. 45 ›› Issue (2): 368-375.DOI: 10.19852/j.cnki.jtcm.2025.02.013
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Regulation of mild moxibustion on non-neuronal cholinergic system in ulcerative colitis rats
PENG Guangbin1, LI Han1, ZHU Lu1, QI Qin1(
), ZHENG Shiyu1, ZHANG Linshan2, LIN Yaying1, MA Zhe1, WU Luyi3, HUANG Yan1, WU Huangan1()
- 1 Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200437, China
2 Shanghai Qigong Research Institute, Shanghai 200030, China
3 Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
-
Received:2024-02-12Accepted:2024-05-20Online:2025-04-15Published:2025-03-10 -
Contact:Prof. WU Huangan, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200437, China. wuhuangan@126.com; MD. QI Qin, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200437, China. 719928895@qq.com, Telephone: +86-13601611239; +86-18616826239 -
Supported by:National Natural Science Foundation of China: Study for the Mechanism of Moxibustion in Ulcerative Colitis based on the α7 Nicotinic Acetylcholine Receptor Mediated Cholinergic Anti-inflammatory Pathway(82205293);National Natural Science Foundation of China: Study of the Central Nervous System Regulatory Mechanism of Moxibustion Repair of Ulcerative Colitis Gut Vascular Barrier Based on Ubiquitin Specific Peptidase 14 Deubiquitination(82274641);National Natural Science Foundation of China: Moxibustion Regulates P300-mediated Histone H3K27 Acetylation Modification in the Treatment of Crohn's Disease(82205262);National Natural Science Foundation of China: Study on the Protective Mechanism of Moxibustion on Intestinal Mucosal Barrier in Ulcerative Colitis based on GABAergic System(82105012);Shanghai Sailing Program: to Study the Protective Effect of Moxibustion on Intestinal Mucosal Barrier in Crohn's Disease Based on Histone H3 Acetylation Modification(22YF1444100);State Administration of Traditional Chinese Medicine High-level Key Discipline Construction Project(zyyzdxk-2023068)
Cite this article
PENG Guangbin, LI Han, ZHU Lu, QI Qin, ZHENG Shiyu, ZHANG Linshan, LIN Yaying, MA Zhe, WU Luyi, HUANG Yan, WU Huangan. Regulation of mild moxibustion on non-neuronal cholinergic system in ulcerative colitis rats[J]. Journal of Traditional Chinese Medicine, 2025, 45(2): 368-375.
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Figure 1 Histopathological manifestations of colonic tissues in rats in each group A: the colonic histopathological manifestations of normal and model rats (hematoxylin-eosin staining, × 200). A1: normal rats; A2: model rats. B: the colonic histopathological manifestations of rats in experiment 1 (hematoxylin-eosin staining, × 200). B1: Con group; B2: Mod group; B3: Mox group; B4: α-BGT group; B5: α-BGT + Mox group. C: the colonic histopathological manifestations of rats in experiment 2 (hematoxylin-eosin staining, × 200). C1: Con group; C2: Mod group; C3: Mox group; C4; VH group; C5: VH + Mox group. D: the colonic histopathological manifestations of rats in experiment 3 (hematoxylin-eosin staining, × 200). D1: Con group; D2: Mod group; D3: Mox group; D4: Qu group; D5: Qu + Mox group. Con group: drink and eat freely + the same fixation as the Mox group; Mod group: drink 4% dextran sodium sulfate for 7 d, and 1% DSS starting from the 8th day + the same fixation as the Mox group; Mox group: mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d; α-BGT: injected with 1 μg/kg α-BGT; α-BGT + Mox: injected with 1 μg/kg α-BGT + mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d; VH: injected with 0.5 mM VH (0.1 mL); VH+ Mox: injected with 0.5 mM VH (0.1 mL) + mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d; Qu group: injected with 100 μM Qu (0.1 mL); Qu + Mox: injected with 100 μM Qu (0.1 mL) + mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d. Con: control group; Mod: UC model group; Mox: mild moxibustion group; α-BGT: alpha-bungarotoxin group; α-BGT + Mox: alpha-bungarotoxin + mild moxibustion group; VH: vesamicol hydrochloride group; VH + Mox: vesamicol hydrochloride + mild moxibustion group; Qu: quinine group; Qu + Mox: quinine + mild moxibustion group. Arrows indicate ulcerated (Red) or healed mucous membranes (Blue).
Figure 1 Histopathological manifestations of colonic tissues in rats in each group A: the colonic histopathological manifestations of normal and model rats (hematoxylin-eosin staining, × 200). A1: normal rats; A2: model rats. B: the colonic histopathological manifestations of rats in experiment 1 (hematoxylin-eosin staining, × 200). B1: Con group; B2: Mod group; B3: Mox group; B4: α-BGT group; B5: α-BGT + Mox group. C: the colonic histopathological manifestations of rats in experiment 2 (hematoxylin-eosin staining, × 200). C1: Con group; C2: Mod group; C3: Mox group; C4; VH group; C5: VH + Mox group. D: the colonic histopathological manifestations of rats in experiment 3 (hematoxylin-eosin staining, × 200). D1: Con group; D2: Mod group; D3: Mox group; D4: Qu group; D5: Qu + Mox group. Con group: drink and eat freely + the same fixation as the Mox group; Mod group: drink 4% dextran sodium sulfate for 7 d, and 1% DSS starting from the 8th day + the same fixation as the Mox group; Mox group: mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d; α-BGT: injected with 1 μg/kg α-BGT; α-BGT + Mox: injected with 1 μg/kg α-BGT + mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d; VH: injected with 0.5 mM VH (0.1 mL); VH+ Mox: injected with 0.5 mM VH (0.1 mL) + mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d; Qu group: injected with 100 μM Qu (0.1 mL); Qu + Mox: injected with 100 μM Qu (0.1 mL) + mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d. Con: control group; Mod: UC model group; Mox: mild moxibustion group; α-BGT: alpha-bungarotoxin group; α-BGT + Mox: alpha-bungarotoxin + mild moxibustion group; VH: vesamicol hydrochloride group; VH + Mox: vesamicol hydrochloride + mild moxibustion group; Qu: quinine group; Qu + Mox: quinine + mild moxibustion group. Arrows indicate ulcerated (Red) or healed mucous membranes (Blue).
Figure 2 Expression of acetylcholine-related molecules in the colon of rats in each group A: representative western blot bands of NF-κB p65 expression in the colon; B: the protein expressions of NF-κB p65 in the colon; C: relative mRNA expression of NF-κB p65 in the colon; D: results of ELISA of ChAT in the colon in experiment 1; E-F: relative mRNA expressions of ChAT and CarAT in the colon in experiment 1, respectively; G-H: results of ELISA of ChAT in the colon in experiment 2 and experiment 3, respectively. Con group: drink and eat freely + the same fixation as the Mox group; Mod group: drink 4% dextran sodium sulfate for 7 d, and 1% DSS starting from the 8th day + the same fixation as the Mox group; Mox group: mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d; α-BGT: injected with 1 μg/kg α-BGT; α-BGT + Mox: injected with 1 μg/kg α-BGT + mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d; VH: injected with 0.5 mM VH (0.1 mL); VH+ Mox: injected with 0.5 mM VH (0.1 mL) + mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d; Qu group: injected with 100 μM Qu (0.1 mL); Qu + Mox: injected with 100 μM Qu (0.1 mL) + mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d. Con: control group; Mod: UC model group; Mox: mild moxibustion group; α-BGT: alpha-bungarotoxin group; α-BGT + Mox: alpha-bungarotoxin + mild moxibustion group; VH: vesamicol hydrochloride group; VH + Mox: vesamicol hydrochloride + mild moxibustion group; Qu: quinine group; Qu + Mox: quinine + mild moxibustion group. ELISA: enzyme-linked immunosorbent assay; NF-κB p65: nuclear factor kappa-B p65 subunit; ChAT: choline acetyltransferase; CarAT: carnosine acetyltransferase. B-D, F-H: data were presented as mean ± standard deviation (in experiment 2, Con group: n = 10, Mod group: n = 8, Mox, VH, and VH + Mox groups: n = 9. Other groups: n = 10). Statistical analyses were measured using one-way analysis of variance followed by least significant difference test for pairwise comparisons. E: data were presented as median (P25, P75) (n = 10). Statistical analyses were measured using the nonparametric Kruskal-Wallis H test. Compared with the control group, aP < 0.01, dP < 0.05; compared with the UC model group, bP < 0.01, fP < 0.05; compared with the mild moxibustion group, cP < 0.01, eP < 0.05; compared with the vesamicol hydrochloride group, gP < 0.05.
Figure 2 Expression of acetylcholine-related molecules in the colon of rats in each group A: representative western blot bands of NF-κB p65 expression in the colon; B: the protein expressions of NF-κB p65 in the colon; C: relative mRNA expression of NF-κB p65 in the colon; D: results of ELISA of ChAT in the colon in experiment 1; E-F: relative mRNA expressions of ChAT and CarAT in the colon in experiment 1, respectively; G-H: results of ELISA of ChAT in the colon in experiment 2 and experiment 3, respectively. Con group: drink and eat freely + the same fixation as the Mox group; Mod group: drink 4% dextran sodium sulfate for 7 d, and 1% DSS starting from the 8th day + the same fixation as the Mox group; Mox group: mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d; α-BGT: injected with 1 μg/kg α-BGT; α-BGT + Mox: injected with 1 μg/kg α-BGT + mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d; VH: injected with 0.5 mM VH (0.1 mL); VH+ Mox: injected with 0.5 mM VH (0.1 mL) + mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d; Qu group: injected with 100 μM Qu (0.1 mL); Qu + Mox: injected with 100 μM Qu (0.1 mL) + mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d. Con: control group; Mod: UC model group; Mox: mild moxibustion group; α-BGT: alpha-bungarotoxin group; α-BGT + Mox: alpha-bungarotoxin + mild moxibustion group; VH: vesamicol hydrochloride group; VH + Mox: vesamicol hydrochloride + mild moxibustion group; Qu: quinine group; Qu + Mox: quinine + mild moxibustion group. ELISA: enzyme-linked immunosorbent assay; NF-κB p65: nuclear factor kappa-B p65 subunit; ChAT: choline acetyltransferase; CarAT: carnosine acetyltransferase. B-D, F-H: data were presented as mean ± standard deviation (in experiment 2, Con group: n = 10, Mod group: n = 8, Mox, VH, and VH + Mox groups: n = 9. Other groups: n = 10). Statistical analyses were measured using one-way analysis of variance followed by least significant difference test for pairwise comparisons. E: data were presented as median (P25, P75) (n = 10). Statistical analyses were measured using the nonparametric Kruskal-Wallis H test. Compared with the control group, aP < 0.01, dP < 0.05; compared with the UC model group, bP < 0.01, fP < 0.05; compared with the mild moxibustion group, cP < 0.01, eP < 0.05; compared with the vesamicol hydrochloride group, gP < 0.05.
Figure 3 Comparison of nAch protein expression in colon tissues of rats in each group in experiment 2 (immunofluorescence staining, × 200) A: Con group. A1: Merge; A2: DAPI; A3: ChAT; A4: PGP9.5. B: Mod group. B1: Merge; B2: DAPI; B3: ChAT; B4: PGP9.5. C: Mox group. C1: Merge; C2: DAPI; C3: ChAT; C4: PGP9.5. D: VH group. D1: Merge; D2: DAPI; D3: ChAT; D4: PGP9.5. E: VH + Mox group. E1: Merge; E2: DAPI; E3: ChAT; E4: PGP9.5. Con group: drink and eat freely + the same fixation as the Mox group; Mod group: drink 4% dextran sodium sulfate for 7 d, and 1% DSS starting from the 8th day + the same fixation as the Mox group; Mox group: mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d; VH: injected with 0.5 mM VH (0.1 mL); VH + Mox: injected with 0.5 mM VH (0.1 mL) + mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d. Con: control group; Mod: UC model group; Mox: mild moxibustion group; VH: vesamicol hydrochloride group; VH + Mox: vesamicol hydrochloride + mild moxibustion group; DAPI: 4',6-diamidino-2-phenylindole; ChAT: choline acetyltransferase; PGP9.5: protein gene product 9.5.
Figure 3 Comparison of nAch protein expression in colon tissues of rats in each group in experiment 2 (immunofluorescence staining, × 200) A: Con group. A1: Merge; A2: DAPI; A3: ChAT; A4: PGP9.5. B: Mod group. B1: Merge; B2: DAPI; B3: ChAT; B4: PGP9.5. C: Mox group. C1: Merge; C2: DAPI; C3: ChAT; C4: PGP9.5. D: VH group. D1: Merge; D2: DAPI; D3: ChAT; D4: PGP9.5. E: VH + Mox group. E1: Merge; E2: DAPI; E3: ChAT; E4: PGP9.5. Con group: drink and eat freely + the same fixation as the Mox group; Mod group: drink 4% dextran sodium sulfate for 7 d, and 1% DSS starting from the 8th day + the same fixation as the Mox group; Mox group: mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d; VH: injected with 0.5 mM VH (0.1 mL); VH + Mox: injected with 0.5 mM VH (0.1 mL) + mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d. Con: control group; Mod: UC model group; Mox: mild moxibustion group; VH: vesamicol hydrochloride group; VH + Mox: vesamicol hydrochloride + mild moxibustion group; DAPI: 4',6-diamidino-2-phenylindole; ChAT: choline acetyltransferase; PGP9.5: protein gene product 9.5.
Figure 4 Comparison of nnAch protein expression in colon tissues of rats in each group in experiment 3 (immunofluorescence staining, × 200) A: Con group. A1: Merge; A2: DAPI; A3: ChAT; A4: OCT1. B: Mod group. B1: Merge; B2: DAPI; B3: ChAT; B4: OCT1. C: Mox group. C1: Merge; C2: DAPI; C3: ChAT; C4: OCT1. D: Qu group. D1: Merge; D2: DAPI; D3: ChAT; D4: OCT1. E: Qu + Mox group. E1: Merge; E2: DAPI; E3: ChAT; E4: OCT1. Con group: drink and eat freely + the same fixation as the Mox group; Mod group: drink 4% dextran sodium sulfate for 7 d, and 1% DSS starting from the 8th day + the same fixation as the Mox group; Mox group: mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d; Qu group: injected with 100 μM Qu (0.1 mL); Qu + Mox: injected with 100 μM Qu (0.1 mL) + mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d. Con: control group; Mod: UC model group; Mox: mild moxibustion group; Qu: quinine group; Qu + Mox: quinine + mild moxibustion group; DAPI: 4',6-diamidino-2-phenylindole; ChAT: choline acetyltransferase; OCT1: organic cation transport 1.
Figure 4 Comparison of nnAch protein expression in colon tissues of rats in each group in experiment 3 (immunofluorescence staining, × 200) A: Con group. A1: Merge; A2: DAPI; A3: ChAT; A4: OCT1. B: Mod group. B1: Merge; B2: DAPI; B3: ChAT; B4: OCT1. C: Mox group. C1: Merge; C2: DAPI; C3: ChAT; C4: OCT1. D: Qu group. D1: Merge; D2: DAPI; D3: ChAT; D4: OCT1. E: Qu + Mox group. E1: Merge; E2: DAPI; E3: ChAT; E4: OCT1. Con group: drink and eat freely + the same fixation as the Mox group; Mod group: drink 4% dextran sodium sulfate for 7 d, and 1% DSS starting from the 8th day + the same fixation as the Mox group; Mox group: mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d; Qu group: injected with 100 μM Qu (0.1 mL); Qu + Mox: injected with 100 μM Qu (0.1 mL) + mild moxibustion at the bilateral Zusanli (ST36) for 10 min per day for 7 d. Con: control group; Mod: UC model group; Mox: mild moxibustion group; Qu: quinine group; Qu + Mox: quinine + mild moxibustion group; DAPI: 4',6-diamidino-2-phenylindole; ChAT: choline acetyltransferase; OCT1: organic cation transport 1.
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