Zheng Xue, Zhi Mu-Jun, Zhao Li-Li, Chen Li, He Zong-Qing, Jing Shuang-Shuang, Wang Qing-Bo
First Affiliated Hospital of Henan University of CM, Zhengzhou 450000, China.
Zhongguo Zhen Jiu. 2022 Jul 12;42(7):773-8. doi: 10.13703/j.0255-2930.20210524-k0001.
To observe the effect of blistering moxibustion on the expression levels of 5-hydroxytyptamine (5-HT) and its receptors of the colon tissue in the mice with visceral hypersensitivity of irritable bowel syndrome (IBS), so as to explore the effect mechanism of blistering moxibustion in treatment of IBS.
Forty SPF-grade newborn Kunming mice were randomly divided into a normal group, a model group, an antagonist group and a blistering moxibustion group, 10 mice in each one. Before modeling, the injection with 0.2 mL parachlorophenylalanine (PCPA) was given on the lateral ventricle in the antagonist group. The endorectal glacial acetic acid stimulation combined with tail clipping was used to prepare the model of visceral hypersensitivity of IBS in the model group, the antagonist group and the blistering moxibustion group. After modeling, in the blistering moxibustion group, the intervention with blistering moxibustion was exerted at "Zhongwan" (CV 12), "Tianshu" (ST 25) and "Zusanli" (ST 36), once herbal irritant plaster at each acupoint, for 2 h each time, once a week, consecutively for 3 weeks. Abdominal withdrawal reflex (AWR) score and electromyographic (EMG) amplitude of abdominal muscles were adopted to evaluate the visceral hypersensitivity. HE staining was applied to observe the morphological changes in colon tissue, and immunohistochemistry was to determine the expression levels of 5-HT and its receptors.
Compared with the normal group, EMG amplitude of abdominal muscles was increased under 20, 40 mm Hg (1 mm Hg=0.133 kPa) in the model group (<0.05), AWR scores and EMG amplitude of abdominal muscles under 60, 80 mm Hg were all increased in the model group (<0.05). In comparison with the model group, EMG amplitude of abdominal muscles was reduced under 20 mm Hg in the blistering moxibustion group (<0.05), AWR scores were increased under 40 mm Hg in both the blistering moxibustion group and the antagonist group (<0.05); AWR scores and EMG amplitude of abdominal muscles under 60, 80 mm Hg were all reduced in both the blistering moxibustion group and the antagonist group (<0.05). Compared with the normal group, in the model group, the mucosa was slightly disturbed, while, the moderate inflammatory cells were visible in the submucosa. In comparison with the model group, the inherent glands of mucosa were regular in shape and a small number of inflammatory cells were visible in both the blistering moxibustion group and the antagonist group. In comparison with the normal group, the average positive staining area percentage (APSAP) of 5-HT and 5-HTR of the colon tissue was increased, while, APSAP of 5-HTR was reduced in the model group (<0.05). Compared with the model group, APSAP of 5-HT and 5-HTR was reduced in both the blistering moxibustion group and the antagonist group (<0.05).
Blistering moxibustion can relieve the visceral hypersensitivity of the mice with visceral hypersensitive IBS and the underlying mechanism is related to the regulation of the gut-brain axis mediated by 5-HT signaling pathway.
观察隔物灸对肠易激综合征（IBS）内脏高敏感小鼠结肠组织5-羟色胺（5-HT）及其受体表达水平的影响，以探讨隔物灸治疗IBS的作用机制。
将40只SPF级新生昆明小鼠随机分为正常组、模型组、拮抗剂组和隔物灸组，每组10只。造模前，拮抗剂组侧脑室注射0.2 mL对氯苯丙氨酸（PCPA）。模型组、拮抗剂组和隔物灸组采用直肠内注入冰醋酸刺激并结合夹尾法制备IBS内脏高敏感模型。造模后，隔物灸组于“中脘”（CV 12）、“天枢”（ST 25）、“足三里”（ST 36）穴进行隔物灸干预，每个穴位贴敷1次中药发泡贴，每次2 h，每周1次，连续3周。采用腹部回撤反射（AWR）评分及腹肌肌电图（EMG）幅值评估内脏高敏感性。采用苏木精-伊红（HE）染色观察结肠组织形态学变化，采用免疫组织化学法检测5-HT及其受体的表达水平。
与正常组比较，模型组在20、40 mmHg（1 mmHg = 0.133 kPa）时腹肌EMG幅值升高（P<0.05），在60、80 mmHg时AWR评分及腹肌EMG幅值均升高（P<0.05）。与模型组比较，隔物灸组在20 mmHg时腹肌EMG幅值降低（P<0.05），隔物灸组和拮抗剂组在40 mmHg时AWR评分升高（P<0.05）；隔物灸组和拮抗剂组在60、80 mmHg时AWR评分及腹肌EMG幅值均降低（P<0.05）。与正常组比较，模型组黏膜轻度紊乱，黏膜下层可见中度炎性细胞。与模型组比较，隔物灸组和拮抗剂组黏膜固有腺体形态规则，可见少量炎性细胞。与正常组比较，模型组结肠组织5-HT及5-羟色胺受体（5-HTR）平均阳性染色面积百分比（APSAP）升高，5-HTR的APSAP降低（P<0.05）。与模型组比较，隔物灸组和拮抗剂组5-HT及5-HTR的APSAP均降低（P<0.05）。
隔物灸可缓解内脏高敏感IBS小鼠的内脏高敏感性，其作用机制可能与调节5-HT信号通路介导的肠-脑轴有关。
