电针通过非神经元胆碱能系统对溃疡性结肠炎小鼠黏膜愈合的影响及机制
Effect and Mechanisms of Electroacupuncture on Mucosal Healing in Ulcerative Colitis Mice via Non-neuroronal Cholinergic System.
作者信息
Li Han, He Wei, Wang Xiao-Yu, Wan Hong-Ye, Chen Li-Zhen, Zhan Li-Bin, Zhang Qi, Jing Xiang-Hong
机构信息
Department of Acupuncture and Moxibustion, Changzhou Affiliated Hospital of Nanjing University of Chinese Medicine, Changzhou, Jiangsu Province, 213002, China.
Research Center of Meridians, Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
出版信息
Chin J Integr Med. 2025 Jun 16. doi: 10.1007/s11655-025-3811-3.
OBJECTIVE
To explore the effect of electroacupuncture (EA) on enhancing local acetylcholine (ACh) synthesis through activation of the nonneuronal cholinergic system (NNCS) and its action on the muscarinic ACh receptor (mAChR)-mediated signalling pathway for repair of mucosal barrier in ulcerative colitis (UC) mice.
METHODS
Twenty-four wild-type male C57BL/6 mice were randomly divided into 4 groups using a random number table method, including control, model, EA, and sham EA (SEA) groups, 6 mice in each group. The UC model mice were induced by 2.5% dextran sodium sulfate (DSS) in free drinking water for 1 week. EA was administered by electrical stimulation (parameters: 2/15 Hz, 0.8 mA, 30 min/d) at the bilateral Zusanli (ST 36) for 7 consecutive days. SEA was performed at the same acupoints without electrical stimulation. The disease activity index (DAI) was determined and colonic permeability were analysed by fluorescein isothiocyanate dextran. The mucosal barrier and tight junctions (TJs) were observed by transmission electron microscopy. Zonula occludens-1 (ZO-1) and colocalization of choline acetyltransferase (ChAT) with organic cation transporters (OCT) and vesicular ACh transporter (VAChT) were tested by immunofluorescence analysis. The protein expressions of myosin light chain (MLC), phosphorylation MLC (pMLC), phospholipase C (PLC), ChAT, nuclear factor kappa-B p65 (NF-κ Bp65), mAChR, OCT and VAChT were measured by Western blot.
RESULTS
Compared with the model group, DAI score, colonic permeability, as well as the protein expressions of pMLC, NF-κ Bp65 and VAChT were decreased in the EA group, and the mucosal barrier and TJs were repaired by EA treatment (P<0.05 or P<0.01). The expressions of ZO-1, mAChR and ChAT in the EA group were increased (P<0.05). The fluorescence intensities of ChAT relative to OCT and the ratio of the colocalization of ChAT with OCT and VAChT were increased as well (P<0.05 or P<0.01).
CONCLUSION
EA at bilateral ST 36 may activate the NNCS, promote ACh release by OCT, activate mAChR-related signaling by inhibiting pMLC expression, upregulate ZO-1, reduce colonic permeability and repair TJs to achieve mucosal barrier repair in UC.
目的
探讨电针（EA）通过激活非神经元胆碱能系统（NNCS）增强局部乙酰胆碱（ACh）合成的作用及其对毒蕈碱型ACh受体（mAChR）介导的信号通路在溃疡性结肠炎（UC）小鼠黏膜屏障修复中的作用。
方法
采用随机数字表法将24只野生型雄性C57BL/6小鼠随机分为4组，即对照组、模型组、电针组和假电针组（SEA组），每组6只。UC模型小鼠通过自由饮用2.5%葡聚糖硫酸钠（DSS）诱导1周。电针组在双侧足三里（ST 36）进行电刺激（参数：2/15 Hz，0.8 mA，30 min/d），连续7天。假电针组在相同穴位进行无电刺激操作。测定疾病活动指数（DAI），并通过异硫氰酸荧光素葡聚糖分析结肠通透性。通过透射电子显微镜观察黏膜屏障和紧密连接（TJs）。通过免疫荧光分析检测紧密连接蛋白1（ZO-1）以及胆碱乙酰转移酶（ChAT）与有机阳离子转运体（OCT）和囊泡型ACh转运体（VAChT）的共定位情况。采用蛋白质印迹法检测肌球蛋白轻链（MLC）、磷酸化MLC（pMLC）、磷脂酶C（PLC）、ChAT、核因子κB p65（NF-κBp65）、mAChR、OCT和VAChT的蛋白表达。
结果
与模型组相比，电针组的DAI评分、结肠通透性以及pMLC、NF-κBp65和VAChT的蛋白表达降低，电针治疗修复了黏膜屏障和TJs（P<0.05或P<0.01）。电针组中ZO-1、mAChR和ChAT的表达增加（P<0.05）。ChAT相对于OCT的荧光强度以及ChAT与OCT和VAChT的共定位比例也增加（P<0.05或P<0.01）。
结论
双侧ST 36电针可能激活NNCS，通过OCT促进ACh释放，通过抑制pMLC表达激活mAChR相关信号，上调ZO-1，降低结肠通透性并修复TJs，从而实现UC小鼠黏膜屏障的修复。
Zotero 插件