College of Acupuncture-moxibustion and Tuina, Chengdu University of Traditional Chinese Medicine, Chengdu 610075, China.
Zhen Ci Yan Jiu. 2024 Feb 25;49(2):145-154. doi: 10.13702/j.1000-0607.20221282.
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.
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.
Compared with the normal group after treatment, the model group showed increased DAI score (<0.001), decreased AWR minimum volume threshold (<0.01), shortened colon length (<0.001), significant inflammatory infiltration in the colon tissue, reduced mucin secretion (<0.01), increased collagen fiber deposition (<0.001), and elevated expression levels of TNF-α, TNF-R1, P38 MAPK, and TRPV1 (<0.001, <0.01, <0.05). Compared with the model group, the MM group showed decreased DAI score (<0.01), increased AWR minimum volume threshold (<0.001), elongated colon length (<0.001), reduced inflammatory cell infiltration, improved integrity of mucosal glandular structure, enhanced mucin secretion (<0.01), decreased collagen fiber deposition (P<0.001), decreased number of mast cells in the colon tissue (<0.001), and decreased expression levels of TNF-α, TNF-R1, P38 MAPK, and TRPV1 (<0.001, <0.01, <0.05). There were no significant differences in the above index between the NM group and the normal group.
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.
观察艾灸“足三里”(ST36)对慢性溃疡性结肠炎(UC)模型小鼠结肠组织中肿瘤坏死因子(TNF)-α、TNF 受体 1(TNF-R1)、p38 丝裂原活化蛋白激酶(P38 MAPK)和瞬时受体电位香草酸 1(TRPV1)表达水平的影响,探讨艾灸改善慢性 UC 内脏高敏性的作用机制。
雄性 C57BL/6J 小鼠随机分为正常组、正常艾灸组(NM 组)、模型组和模型艾灸组(MM 组),每组 10 只。采用饮用 2.5%葡聚糖硫酸钠溶液的方法复制慢性 UC 模型,共 3 个周期,每周 5 天,休息 2 天。NM 组和 MM 组于造模第 1 天开始艾灸 ST36,每次 20 min,每周 5 天,共 4 周。治疗前后评估各组疾病活动指数(DAI)评分,测量腹壁回缩反射(AWR)最小容积阈值以观察小鼠肠敏感性,测量结肠长度,HE 染色观察结肠组织病理变化,过碘酸雪夫染色观察结肠杯状细胞黏蛋白分泌情况,Masson 染色观察肠纤维化,免疫荧光染色检测结肠组织中胰蛋白酶阳性细胞(即肥大细胞)数和 TNF-α表达水平,免疫组化检测结肠组织中 TNF-R1、P38 MAPK 和 TRPV1 表达水平。
与正常组比较,模型组 DAI 评分升高(P<0.001),AWR 最小容积阈值降低(P<0.01),结肠长度缩短(P<0.001),结肠组织炎症浸润明显,黏蛋白分泌减少(P<0.01),胶原纤维沉积增加(P<0.001),TNF-α、TNF-R1、P38 MAPK 和 TRPV1 表达水平升高(P<0.001,P<0.01,P<0.05);与模型组比较,MM 组 DAI 评分降低(P<0.01),AWR 最小容积阈值升高(P<0.001),结肠长度延长(P<0.001),炎症细胞浸润减少,肠黏膜腺结构完整性改善,黏蛋白分泌增加(P<0.01),胶原纤维沉积减少(P<0.001),结肠组织中肥大细胞数减少(P<0.001),TNF-α、TNF-R1、P38 MAPK 和 TRPV1 表达水平降低(P<0.001,P<0.01,P<0.05)。NM 组与正常组比较,上述各指标差异均无统计学意义。
艾灸可降低慢性 UC 模型小鼠内脏高敏性,减轻结肠组织炎症浸润和纤维化损伤,其作用机制可能与下调 TNF-α、TNF-R1、P38 MAPK 和 TRPV1 的表达有关。