针刺对腹泻型肠易激综合征大鼠神经肽及相关炎症因子表达的影响。

Effect of acupuncture on the expression of neuropeptides and related inflammatory factors in rats with diarrhea-predominant irritable bowel syndrome.

机构信息

Acupuncture-moxibustion and Tuina College, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, China.

出版信息

Zhen Ci Yan Jiu. 2023 Nov 25;48(11):1142-1150. doi: 10.13702/j.1000-0607.20221401.

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(<0.05, <0.01), increased fecal pellet number (<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(<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(<0.01, <0.05), significantly decreased fecal pellet number (<0.05), significantly decreased MC number in the colon tissue, serum TNF-α contents, and CGRP mRNA expressions in the hypothalamus and colon tissue(<0.01);at the same time, the acupuncture group showed significantly decreased CRF expressions in the hypothalamus and colon tissue(<0.01, <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.

摘要

目的

观察针刺对腹泻型肠易激综合征(IBS-D)大鼠神经肽及相关炎症因子表达的影响,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。除空白组外,其余各组采用醋酸灌肠联合束缚应激的方法连续造模 14 天,建立 IBS-D 大鼠模型。造模成功后,药物组给予溴匹斯的明(15mg/kg)灌胃,每日 1 次,针刺组针刺“百会”(GV20)和双侧“天枢”(ST25)、“上巨虚”(ST37)、“足三里”(ST36)、“太冲”(LR3),每日 20min,连续治疗 14 天。观察各组大鼠一般状态,测量大鼠体重,进行旷场实验,测量大鼠水平和垂直运动以及粪便颗粒数,HE 染色观察大鼠下丘脑和结肠组织的病理形态学改变,甲苯胺蓝染色观察并计数大鼠结肠组织中的肥大细胞(MC),ELISA 法检测大鼠血清肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)含量,实时荧光定量 PCR 法检测大鼠下丘脑和结肠组织中降钙素基因相关肽(CGRP)mRNA 表达,Western blot 法检测大鼠下丘脑和结肠组织中促肾上腺皮质释放激素(CRF)表达。

结果

HE 染色显示,模型组大鼠结肠固有层有炎症细胞浸润,各治疗组均有减轻。与空白组比较,模型组大鼠体重明显降低(P<0.01),旷场实验中水平运动距离和直立次数明显减少(P<0.01),血清 IL-10 含量明显降低(P<0.01,P<0.05),粪便颗粒数明显增多(P<0.01),结肠组织 MC 数、血清 TNF-α含量、下丘脑和结肠组织 CGRP mRNA 表达及 CRF 表达均明显升高(P<0.01);与模型组比较,药物组和针刺组大鼠体重明显增加(P<0.01,P<0.05),旷场实验中水平运动距离和直立次数明显增加(P<0.01,P<0.05),血清 IL-10 含量明显升高(P<0.01,P<0.05),粪便颗粒数明显减少(P<0.05),结肠组织 MC 数、血清 TNF-α含量、下丘脑和结肠组织 CGRP mRNA 表达及 CRF 表达均明显降低(P<0.01);针刺组大鼠下丘脑和结肠组织 CRF 表达明显降低(P<0.01,P<0.05)。药物组与针刺组比较,上述各指标差异均无统计学意义。

结论

针刺可改善 IBS-D 大鼠一般状态和情绪,调节炎症反应和神经肽表达,缓解 IBS-D 症状,其作用机制可能与调节神经肽和炎症因子水平有关。

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