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Title: Effects of moxibustion on visceral hypersensitivity and colonic inflammatory response in mice with chronic ulcerative colitis.
Author: Chen W, Cheng B, Yu SG, Wu QF.
Journal: Zhen Ci Yan Jiu; 2024 Feb 25; 49(2):145-154. PubMed ID: 38413035.
Abstract:
OBJECTIVES: To observe the effects of moxibustion at "Zusanli" (ST36) on the expression levels of tumor necrosis factor (TNF)-α, TNF receptor 1 (TNF-R1), p38 mitogen-activated protein kinase (P38 MAPK), and transient receptor potential vanilloid 1 (TRPV1) in the colon tissue of mice with chronic ulcerative colitis (UC), so as to explore the underlying mechanisms of moxibustion in improving visceral hypersensitivity in chronic UC. METHODS: Male C57BL/6J mice were randomly divided into normal group, normal with moxibustion (NM) group, model group, and model with moxibustion (MM) group, with 10 mice in each group. The chronic UC model was established by drinking 2.5% dextran sodium sulfate for 3 cycles. Mice in the NM and MM groups received moxibustion at ST36 for 20 min, 5 days per week with a 2-day break, for a total of 4 weeks. The disease activity index (DAI) score of each group was evaluated before and after treatment. The minimum volume threshold of abdominal wall retraction reflex (AWR) was measured to observe the intestinal sensitivity of mice. The colon length was measured. The pathological changes of colon tissue were observed by HE staining. The expression of mucin in colon goblet cells was detected by periodate Scheff staining. The intestinal fibrosis was observed by Masson staining. The number of trypsin-positive cells (i.e., mast cell) and the expression level of TNF-α in colon tissue were detected by immunofluorescence staining. The expression levels of TNF-R1, P38 MAPK and TRPV1 in colon tissue were detected by immunohistochemistry. RESULTS: Compared with the normal group after treatment, the model group showed increased DAI score (P<0.001), decreased AWR minimum volume threshold (P<0.01), shortened colon length (P<0.001), significant inflammatory infiltration in the colon tissue, reduced mucin secretion (P<0.01), increased collagen fiber deposition (P<0.001), and elevated expression levels of TNF-α, TNF-R1, P38 MAPK, and TRPV1 (P<0.001, P<0.01, P<0.05). Compared with the model group, the MM group showed decreased DAI score (P<0.01), increased AWR minimum volume threshold (P<0.001), elongated colon length (P<0.001), reduced inflammatory cell infiltration, improved integrity of mucosal glandular structure, enhanced mucin secretion (P<0.01), decreased collagen fiber deposition (P<0.001), decreased number of mast cells in the colon tissue (P<0.001), and decreased expression levels of TNF-α, TNF-R1, P38 MAPK, and TRPV1 (P<0.001, P<0.01, P<0.05). There were no significant differences in the above index between the NM group and the normal group. CONCLUSIONS: Moxibustion can reduce visceral hypersensitivity, alleviate inflammatory infiltration and fibrotic damage in the colon tissue of mice with chronic UC. These effects may be associated with the down-regulation of TNF-α, TNF-R1, P38 MAPK, and TRPV1 expression in colon. 目的: 观察艾灸“足三里”对慢性溃疡性结肠炎（UC）小鼠结肠组织中肿瘤坏死因子-α（TNF-α）、肿瘤坏死因子受体1（TNF-R1）、P38丝裂原活化蛋白激酶（P38 MAPK）、瞬时感受器电位香草酸受体1（TRPV1）表达的影响，探讨艾灸改善慢性UC内脏高敏的可能机制。方法: 雄性C57BL/6J小鼠按体质量随机分为正常组、正常+艾灸组、模型组、模型+艾灸组，每组10只。以2.5%的葡聚糖硫酸钠循环3个周期饮用法建立慢性UC模型。正常+艾灸组和模型+艾灸组小鼠予以“足三里”艾灸，每次20 min，每周连续干预5 d，休息2 d，共4周。评价各组小鼠治疗前后的疾病活动指数（DAI）评分；测定各组小鼠腹壁回撤反射（AWR）最小容量阈值以观察小鼠肠道敏感性；测量各组小鼠结肠长度；HE染色观察各组小鼠结肠组织病理变化；过碘酸雪夫染色检测各组小鼠结肠组织杯状细胞中的黏液素分泌；Masson染色观察各组小鼠肠道纤维化情况；免疫荧光染色法检测各组小鼠结肠组织类胰蛋白酶阳性细胞数（即肥大细胞数）及TNF-α表达水平；免疫组织化学法检测各组小鼠结肠组织TNF-R1、P38 MAPK、TRPV1表达水平。结果: 艾灸治疗后，与正常组相比，模型组小鼠DAI评分升高（P<0.001），AWR最小容量阈值降低（P<0.01），结肠长度缩短（P<0.001），结肠组织炎性浸润明显，深入基底层，黏液素分泌减少（P<0.01），胶原纤维增加（P<0.001），结肠组织中肥大细胞数量增加（P<0.001），TNF-α、TNF-R1、P38 MAPK、TRPV1表达水平均明显升高（P<0.001，P<0.01，P<0.05）。与模型组相比，模型+艾灸组小鼠DAI评分降低（P<0.01），AWR最小容量阈值升高（P<0.001），结肠长度增加（P<0.001），炎性细胞浸润减少，黏膜肠腺结构完整性增加，黏液素分泌增多（P<0.01），胶原纤维减少（P<0.001），结肠组织中肥大细胞数量减少（P<0.001），TNF-α、TNF-R1、P38 MAPK、TRPV1表达水平均明显降低（P<0.001，P<0.01，P<0.05）。正常+艾灸组小鼠各项指标与正常组相比差异无统计学意义。结论: 艾灸可减轻慢性UC小鼠内脏高敏，减轻肠道肥大细胞炎性浸润及纤维化损伤，该效应可能与艾灸下调结肠TNF-α、TNF-R1、P38 MAPK、TRPV1表达有关。.

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