Anhui University of Chinese Medicine, Hefei 230038, China.
The Second Affiliated Hospital of Anhui University of Chinese Medicine, Hefei 230061.
Zhen Ci Yan Jiu. 2024 Jan 25;49(1):47-56. doi: 10.13702/j.1000-0607.20221092.
To observe the effect of moxibustion intervention on the hypothalamus-spinal cord-colon axis of rats with irritable bowel syndrome with diarrhea (IBS-D) and explore the mechanism of moxibustion in improving visceral hypersensitivity in rats with IBS-D.
A total of 36 SD rats were randomly divided into normal, model, and moxibustion groups, with 12 rats in each group. The IBS-D model was established by maternal separation + acetic acid stimulation + chronic restraint. Rats of the moxibustion group received bilateral moxibustion on "Tianshu" (ST25) and "Shangjuxu" (ST37) for 15 min, once a day for 7 consecutive days. The body weight, loose stool rate, and minimum threshold volume of abdominal withdrawal reflex (AWR) were measured before and after moxibustion intervention, respectively. The histopathological changes in the colon tissue were observed after HE staining. The number of colonic mucosal mast cells (MCs) was measured by toluidine blue staining. The activation of MCs was determined by tryptase positive expression level and examined by immunohistochemical staining. The content, protein and mRNA expression levels and positive expression levels of corticotropin releasing factor (CRF), substance P (SP), and calcitonin gene-related peptide (CGRP) in the hypothalamus, spinal cord and colon tissues were measured by ELISA, Western blot, real-time fluorescent quantitative PCR and immunofluorescence staining, respectively.
Compared with the normal group, the loose stool rate was increased (<0.01)；the body weight and minimum threshold volume of AWR were decreased (<0.01)；the inflammatory infiltration of colon tissues was obvious；the number of MCs and positive expression level of tryptase in the colon tissue were increased (<0.01)；the contents, positive expression le-vels, protein and mRNA expression levels of CRF, SP and CGRP in the hypothalamus, spinal cord and colon tissues were increased (<0.01, <0.05) in the model group. After the intervention, compared with the model group, all these indicators showed opposite trends (<0.01, <0.05) in the moxibustion group.
Moxibustion can improve visceral hypersensitivity in rats with IBS-D, and its mechanism may be related to regulating the hypothalamic-spinal-colon axis to reduce the release of CRF, SP and CGRP, and thus to inhibite MC in colon tissue.
观察艾灸干预对腹泻型肠易激综合征（IBS-D）大鼠下丘脑-脊髓-结肠轴的影响，探讨艾灸改善 IBS-D 大鼠内脏高敏性的机制。
36 只 SD 大鼠随机分为正常组、模型组和艾灸组，每组 12 只。采用母婴分离+醋酸刺激+慢性束缚的方法建立 IBS-D 模型。艾灸组大鼠双侧“天枢”（ST25）和“上巨虚”（ST37）艾灸 15min，每日 1 次，连续 7 天。艾灸干预前后分别测量大鼠体质量、稀便率和腹壁反射最小阈值容积（AWR）。HE 染色观察结肠组织的病理变化，甲苯胺蓝染色法测量结肠黏膜肥大细胞（MC）数量，免疫组织化学染色法检测 MC 脱颗粒水平（以胰蛋白酶阳性表达水平表示）。采用 ELISA 法、Western blot 法、实时荧光定量 PCR 法和免疫荧光染色法分别检测下丘脑、脊髓和结肠组织中促肾上腺皮质释放因子（CRF）、P 物质（SP）和降钙素基因相关肽（CGRP）的含量、蛋白和 mRNA 表达水平及其阳性表达水平。
与正常组比较，模型组大鼠稀便率升高（P<0.01），体质量和 AWR 最小阈值容积降低（P<0.01），结肠组织炎症浸润明显，结肠组织 MC 数量和胰蛋白酶阳性表达水平升高（P<0.01），下丘脑、脊髓和结肠组织中 CRF、SP 和 CGRP 的含量、阳性表达水平、蛋白和 mRNA 表达水平均升高（P<0.01，P<0.05）。与模型组比较，艾灸组上述指标均呈现出相反的变化趋势（P<0.01，P<0.05）。
艾灸可以改善 IBS-D 大鼠内脏高敏性，其机制可能与调节下丘脑-脊髓-结肠轴，减少 CRF、SP 和 CGRP 的释放，从而抑制结肠组织中的 MC 有关。
