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Title: Effect of acupuncture on the expression of neuropeptides and related inflammatory factors in rats with diarrhea-predominant irritable bowel syndrome.
Author: Zhu Z, Yang XF, Yu HM, Chen PB, Jin LM, Li Y, Han D, Shang XM.
Journal: Zhen Ci Yan Jiu; 2023 Nov 25; 48(11):1142-1150. PubMed ID: 37984912.
Abstract:
OBJECTIVES: To observe the effect of acupuncture on the expressions of neuropeptides and related inflammatory factors in rats with diarrhea-predominant irritable bowel syndrome(IBS-D), so as to explore the mechanism of acupuncture in the treatment of IBS-D. METHODS: Male Wistar rats were randomly divided into blank group, model group, medication group, and acupuncture group, with 6 rats in each group. Except for the blank group, the other groups were subjected to 14-day "acetic acid enema + restraint stress" to establish the IBS-D rat model. After successful modeling, the medication group received gavage of pinaverium bromide(15 mg/kg) once a day, and the acupuncture group received acupuncture at "Baihui"(GV20) and bilateral "Tianshu"(ST25), "Shangjuxu"(ST37), "Zusanli"(ST36), and "Taichong"(LR3) for 20 min every day, both groups were treated continuously for 14 days. The general state of the rats in each group was observed, and the body weight of the rats was measured. The open-field experiment was conducted to measure the horizontal and vertical movements, and the number of fecal pellets of rats. The histopathological morphology of hypothalamus and colon of rats was observed by HE staining. Toluidine blue staining was used to observe and count the mast cells(MCs) in the colon tissue of rats. ELISA was used to detect the serum contents of tumor necrosis factor-α(TNF-α) and interleukin(IL)-10. Real-time fluorescence quantitative PCR was performed to detect the mRNA expressions of calcitonin gene-related peptide(CGRP) in the hypothalamus and colon tissue. Western blot was used to detect the expressions of corticotropin-releasing factor(CRF) in the hypothalamus and colon tissue. RESULTS: HE staining showed that there was inflammatory cell infiltration in the lamina propria of colon in the model group, and it was reduced in the other groups. Compared with the blank group, the model group showed significantly decreased body weight, decreased walking distance and upright times in open field experiment, decreased serum IL-10 contents(P<0.05, P<0.01), increased fecal pellet number (P<0.01), increased MC number in the colon tissue, serum TNF-α contents, and CGRP mRNA expressions and CRF expressions in the hypothalamus and colon tissue(P<0.01). Compared with the model group, both medication and acupuncture groups showed significantly increased body weight, walking distance and upright times in the open-field experiment, and serum IL-10 contents(P<0.01, P<0.05), significantly decreased fecal pellet number (P<0.05), significantly decreased MC number in the colon tissue, serum TNF-α contents, and CGRP mRNA expressions in the hypothalamus and colon tissue(P<0.01)；at the same time, the acupuncture group showed significantly decreased CRF expressions in the hypothalamus and colon tissue(P<0.01, P<0.05). There was no significant difference in the above indicators between the medication group and the acupuncture group. CONCLUSIONS: Acupuncture can improve the general and emotional state, inflammatory response, and neuropeptide expression in rats with IBS-D, and alleviate the symptoms of IBS-D, which may be related to the regulation of neuropeptides and inflammatory factors levels. 目的: 观察针刺对腹泻型肠易激综合征（IBS-D）大鼠脑肠肽及相关炎性因子表达的影响，探讨针刺治疗IBS-D的作用机制。方法: 雄性Wistar大鼠随机分为空白组、模型组、药物组和针刺组，每组6只。除正常组外，其余各组采用“冰醋酸灌肠+束缚应激”方式建立IBS-D大鼠模型，共14 d。造模成功后药物组予匹维溴铵（15 mg/kg）灌胃，1次/d；针刺组予针刺“百会”及双侧“天枢”“上巨虚”“足三里”“太冲”，20 min/d，两组均连续治疗14 d。观察各组大鼠一般状态，测定大鼠体质量，旷场实验测定大鼠横向运动行走距离、纵向运动直立次数和粪便粒数，HE染色法观察大鼠下丘脑及结肠组织病理形态，甲苯胺蓝染色法观察大鼠结肠组织肥大细胞（MC）并计数，ELISA法检测血清肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-10含量，实时荧光定量PCR法检测下丘脑、结肠组织降钙素基因相关肽（CGRP）mRNA表达；Western blot法检测下丘脑、结肠组织促肾上腺皮质激素释放因子（CRF）表达。结果: HE染色示，模型组大鼠结肠固有层有炎性细胞浸润，其余各组有所减轻。与空白组比较，模型组大鼠体质量，旷场实验行走距离及直立次数，血清IL-10含量均显著降低（P<0.05，P<0.01），粪便粒数及结肠组织MC数量显著增加（P<0.01），血清TNF-α含量，下丘脑、结肠组织CGRP mRNA、CRF表达显著升高（P<0.01）。与模型组比较，药物组、针刺组大鼠体质量，旷场实验行走距离及直立次数，血清IL-10含量显著升高（P<0.01，P<0.05）、粪便粒数及结肠组织MC数量显著减少（P<0.05，P<0.01），血清TNF-α含量，下丘脑、结肠组织中CGRP mRNA表达水平显著降低（P<0.01）；同时针刺组下丘脑、结肠组织CRF表达水平显著降低（P<0.01，P<0.05）。以上指标药物组和针刺组两组间比较差异无统计学意义。结论: 针刺可改善IBS-D大鼠一般状态及情绪状态、炎性反应及相关脑肠肽表达，减轻IBS-D症状，其机制可能与调控脑肠肽进而影响炎性因子水平有关。.

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