Zhou Xu-Liu, Wang Hua, Li Jia, Wu Song, Wu Fan, Lu Wei, Fu Yi-Meng
College of Acupuncture-moxibustion and Orthopedics-traumatology, Hubei University of Chinese Medicine, Wuhan 430061, China.
Zhen Ci Yan Jiu. 2023 Jul 25;48(7):635-42. doi: 10.13702/j.1000-0607.20220711.
To observe the effect of acupuncture and moxibustion of the combined "Biao-Ben" acupoints on autonomic nervous function and related factors in rats with irritable bowel syndrome diarrhea (IBS-D).
Female SD rats were randomly divided into normal control, IBS-D model, "Biao-Ben" acupoint combination, and conventional acupoint combination groups, with 10 rats in each group. The IBS-D model was established by repeated chronic stress stimulation (water or food deprivation, painful tail pinching, exposure to a 43 ℃ environment, forced swimming in 4 ℃ water, day-night inversion and horizontal vibration) for 28 d, and followed by acute restraint stress (wrapping of shoulders, forelimbs and trunk) for 1 h and gavage of senna fluid, once daily for 28 d. For rats of the "Biao-Ben" acupoint combination group, acupuncture and moxibustion were applied to "Guanyuan" (CV4), bilateral "Zusanli" (ST36), and "Neiguan" (PC6), and for rats of the conventional acupoint combination group, acupuncture and moxibustion were applied to bilateral ST36, "Tianshu"(ST25), and "Shangjuxu"(ST37). Both acupuncture and suspension moxibustion treatment were conduced for 15 min, once daily for 21 days. The fecal water content was calculated, and the spontaneous activity behaviors (total distance of crossing and the number of squares crossed in 5 min) were evaluated by open-field tests. The abdominal withdrawal reflex (AWR) was detected. H.E. staining was used to observe the histopathological changes of colon tissue. The domains of heart rate variability (HRV) including low frequency (LF), high frequency (HF), and LF/HF were analyzed using an electrophysiological recorder. The contents of calcitonin gene-related peptide (CGRP) in serum and noradrenaline (NE) in plasma were detected using ELISA. The expression level and immunoactivity of 5-hydroxytryptamine (5-HT) in colon tissues were detected by Western blot and immunofluorescence histochemistry, separately.
Compared with the normal group, the model group had a striking increase in fecal water content, AWR scores at 20, 40, 60, and 80 mm Hg, NE and CGRP contents, LF, LF/HF, and 5-HT protein expression and immunoactivity (<0.01), and an obvious decrease in the total distance of crossing, the number of squares crossed, and HF of HRV (<0.01). In comparison with the model group, the fecal water content, AWR scores at 20, 40, 60, and 80 mm Hg, NE and CGRP contents, LF, LF/HF, and 5-HT protein expression and immunoactivity were significantly decreased (<0.01,<0.05), while the total distance of cros-sing, number of squares crossed, HF of HRV were considerably increased (<0.01, <0.05) in both "Biao-Ben" acupoint combination and conventional acupoint combination groups. The effects of the "Biao-Ben" acupoint combination were apparently superior to those of conventional acupoint combination in down-regulating the fecal water content, AWR score at 20, 40, 60 and 80 mm Hg, NE and CGRP contents, LF, LF/HF, and 5-HT expression and immunoactivity, and in increasing the number of squares crossed, and HF of HRV (<0.01, <0.05). HE staining showed no pathological changes in colonic mucosa in all groups.
Acupuncture and moxibustion stimulation of combined "Biao-Ben" acupoints can effectively improve the symptoms (spontaneous activities, visceral hypersensitivity) of IBS-D model rats, which may be related to its functions in regulating autonomic nervous activities, and down-regulating blood NE and CGRP contents and colonic 5-HT protein expression and immunoactivity, and the effects of "Biao-Ben" acupoint combination are superior to those of conventional acupoint combination.
观察针刺与艾灸联合运用“标本”穴位对腹泻型肠易激综合征（IBS-D）大鼠自主神经功能及相关因子的影响。
将雌性SD大鼠随机分为正常对照组、IBS-D模型组、“标本”穴位组合组和传统穴位组合组，每组10只。通过重复慢性应激刺激（禁食或禁水、夹尾致痛、暴露于43℃环境、4℃水中强迫游泳、昼夜颠倒及水平振动）28天建立IBS-D模型，随后进行急性束缚应激（肩部、前肢及躯干包裹）1小时，并灌胃番泻叶液，每日1次，共28天。“标本”穴位组合组大鼠针刺与艾灸“关元”（CV4）、双侧“足三里”（ST36）及“内关”（PC6），传统穴位组合组大鼠针刺与艾灸双侧ST36、“天枢”（ST25）及“上巨虚”（ST37）。针刺与悬灸治疗均为15分钟，每日1次，共21天。计算粪便含水量，通过旷场试验评估自发活动行为（5分钟内穿越的总距离及穿越的方格数）。检测腹部退缩反射（AWR）。采用苏木精-伊红（H.E.）染色观察结肠组织的组织病理学变化。使用电生理记录仪分析心率变异性（HRV）的低频（LF）、高频（HF）及LF/HF等指标。采用酶联免疫吸附测定（ELISA）法检测血清中降钙素基因相关肽（CGRP）含量及血浆中去甲肾上腺素（NE）含量。分别通过蛋白质免疫印迹法（Western blot）和免疫荧光组织化学法检测结肠组织中5-羟色胺（5-HT）的表达水平及免疫活性。
与正常组相比，模型组粪便含水量、20、40、60及80 mmHg时的AWR评分、NE及CGRP含量、LF、LF/HF以及5-HT蛋白表达和免疫活性显著升高（<0.01），旷场试验中穿越的总距离、穿越的方格数及HRV的HF显著降低（<0.01）。与模型组相比，“标本”穴位组合组和传统穴位组合组的粪便含水量、20、40、60及80 mmHg时的AWR评分、NE及CGRP含量、LF、LF/HF以及5-HT蛋白表达和免疫活性均显著降低（<0.01，<0.05），而旷场试验中穿越的总距离、穿越的方格数及HRV的HF均显著增加（<0.01，<0.05）。“标本”穴位组合组在降低粪便含水量、20、40、60及80 mmHg时的AWR评分、NE及CGRP含量、LF、LF/HF以及5-HT表达和免疫活性，以及增加穿越的方格数和HRV的HF方面，其效果明显优于传统穴位组合组（<0.01，<0.05）。H.E.染色显示各组结肠黏膜均无病理改变。
针刺与艾灸联合刺激“标本”穴位能有效改善IBS-D模型大鼠的症状（自发活动、内脏高敏感性），这可能与其调节自主神经活动、降低血液中NE和CGRP含量以及结肠5-HT蛋白表达和免疫活性的作用有关，且“标本”穴位组合的效果优于传统穴位组合。
