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张之涵,刘羽彤,顾玲,赵旭宇,花炀,葛飞,陈美娟.基于5-HT探讨电针刺激天枢、足三里对IBS-D大鼠的影响[J].湖南中医药大学学报,2023,43(12):2264-2271[点击复制] |
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| 基于5-HT探讨电针刺激天枢、足三里对IBS-D大鼠的影响 |
| 张之涵,刘羽彤,顾玲,赵旭宇,花炀,葛飞,陈美娟 |
| (南京中医药大学医学院, 江苏 南京 210023;南京中医药大学附属海安市中医院, 江苏 海安 226600) |
| 摘要: |
| 目的 观察电针刺激天枢、足三里对腹泻型肠易激综合征（diarrheal irritable bowel syndrome，IBS-D）模型大鼠血清5-羟色胺（5-hydroxytryptamine，5-HT）含量及对结肠5-羟色胺受体3A型（5-hydroxytryptamine 3A receptor，5-HT3AR）蛋白表达量的影响，探讨“同经异腑合募配穴”指导下，电针刺激天枢、足三里治疗IBS-D大鼠的作用机制，为电针的临床应用提供实验依据。方法 将24只SD雄性大鼠随机分为空白组、模型组、阿洛司琼组及电针组，每组6只。除空白组外，其余3组大鼠采用慢性束缚、避水应激及灌服番泻叶煎剂相结合的方法建立IBS-D大鼠模型。造模成功后，对电针组予以电针刺激双侧天枢、足三里，15 min/次；空白组、模型组和电针组均给予1 mL/100 g体质量生理盐水灌胃；阿洛司琼组给予1 mg/mL浓度的阿洛司琼溶液按1 mL/100 g体质量灌胃；4组均治疗1次/d，连续治疗14 d。观察大鼠一般情况；通过腹壁回撤反射（abdominal wall retraction reflex，AWR）实验进行内脏敏感性评估；记录大鼠体质量增长率的变化；计算大鼠粪便含水量；HE染色观察结肠组织形态；ELISA法检测血浆中的5-HT含量；Western blot检测目的蛋白5-HT3AR相对表达量。结果 （1）大鼠一般情况：与模型组比较，电针组大鼠精神良好，活动正常，大便干湿适中，为成形粪粒。（2）体质量增长率：与治疗前比较，治疗后空白组、模型组、阿洛司琼组及电针组体质量增长率显著降低（P<0.001）；与空白组比较，模型组与阿洛司琼组体质量增长率降低（P<0.01，P<0.05）；与模型组、阿洛司琼组比较，电针组体质量增长率升高（P<0.05）。（3）粪便含水量：与治疗前比较，电针组粪便含水量显著降低（P<0.01）；与空白组比较，模型组、阿洛司琼组粪便含水量高（P<0.001，P<0.05）；与模型组、阿洛司琼组比较，电针组的粪便含水量降低（P<0.01，P<0.05）。（4）肠道容量阈值：与空白组比较，模型组肠道容量阈值显著降低（P<0.001）；与模型组比较，阿洛司琼组与电针组肠道容量阈值升高（P<0.05）。（5）组织HE染色：与模型组比较，阿洛司琼组黏膜上皮完整，坏死范围明显减小；电针组黏膜上皮完整，吸收细胞呈柱状，黏膜皱壁中可见淋巴细胞和少量中性粒细胞等炎细胞浸润。与阿洛司琼组比较，电针组上皮完整，无坏死炎性损伤程度轻。（6）5-HT通路检测：与空白组比较，模型组血清中5-HT的含量上升（P<0.05）；与模型组比较，电针组血清中5-HT含量下降（P<0.05）。（7）结肠5-HT3AR蛋白表达：与空白组比较，模型组结肠的5-HT3AR蛋白表达升高（P<0.05）；与模型组比较，阿洛司琼组和电针组结肠的5-HT3AR蛋白表达均显著降低（P<0.001）；与阿洛司琼组比较，电针组结肠5-HT3AR蛋白表达显著降低（P<0.001）。结论 在“同经异腑合募配穴”指导下，电针刺激天枢、足三里对IBS-D大鼠有显著的治疗效果，其作用机制可能通过降低血清5-HT含量及结肠5-HT3AR蛋白表达，恢复肠道运动和内脏感觉功能，从而缓解IBS-D的临床症状。 |
| 关键词: 腹泻型肠易激综合征 同经异腑合募配穴 内脏高敏感性 5-HT 5-HT3AR |
| DOI：10.3969/j.issn.1674-070X.2023.12.017 |
| 投稿时间：2023-09-09 |
| 基金项目:国家自然科学基金项目（81503374）；江苏省自然科学基金项目（BK20181235）；江苏省创新训练项目（202210315047Y）。 |
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| Effects of electroacupuncture stimulation of “Tianshu”(ST25) and “Zusanli”(ST36) on IBS-D rats based on 5-HT |
| ZHANG Zhihan,LIU Yutong,GU Ling,ZHAO Xuyu,HUA Yang,GE Fei,CHEN Meijuan |
| (School of Medicine, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, China;Hai'an Hospital of Chinese Medicine affiliated to Nanjing University of Chinese Medicine, Hai'an, Jiangsu 226600, China) |
| Abstract: |
| Objective To observe the effects of electroacupuncture at "Tian shu"(ST25) and "Zusanli"(ST36) on 5-hydroxytryptamine(5-HT) content in serum and 5-hydroxytryptamine 3A receptor(5-HT3AR) protein expression in colon of model rats with diarrheal irritable bowel syndrome(IBS-D), and to explore the mechanism of action of treating IBS-D rats by electroacupuncture stimulation of "Tianshu"(ST25) and "Zusanli"(ST36) under the guidance of "combining he and mu points from the same type of meridians of different fu-organs", thus providing experimental evidence for the clinical application of electroacupuncture.Methods Twenty-four SD male rats were randomized into blank group, model group, alosetron group, and electroacupuncture group, with six rats in each group. Except the blank group, the other three groups of rats were used to establish IBS-D rat models by the combination of chronic restraint, water avoidance stress, and oral administration of Fanxieye(Sennae Folium) decoction. After successful modeling, the electroacupuncture group was given electroacupuncture at bilateral "Tianshu"(ST25) and "Zusanli"(ST36), 15 min each time.The blank group, model group, and electroacupuncture group were administered 1 m L/100 g normal saline by gavage according to the body mass of rats, while the alosetron group was given 1 mg/mL concentration of alosetron solution by gavage at a dose of 1mL/100 g based on body mass. All the four groups were treated once a day for 14 consecutive days. The general condition of rats was observed. Visceral sensitivity was evaluated by abdominal wall retraction reflex(AWR) experiment. In addition, the changes of body mass growth rate were recorded and the fecal water content was calculated. Besides, the morphology of colonic tissues was observed by HE staining, the 5-HT content in plasma was determined by ELISA, and the relative expression of target protein 5-HT3AR was measured by Western blot. Results(1) General condition of rats:Compared with the model group, the rats in electroacupuncture group were in good spirit with normal activity, and the stool were moderately dry and wet with well-formed faecal pellets.(2) The growth rate of body mass:Compared with that before treatment, the growth rate of body mass in the blank group, model group, alosetron group, and electroacupuncture group decreased significantly after treatment(P<0.001). Additionally, compared with the blank group, the growth rate of body mass in model group and alosetron group decreased(P<0.01, P<0.05).Besides, compared with the model group and alosetron group, the growth rate of body mass in electroacupuncture group increased(P<0.05).(3) Fecal water content:Compared with that before treatment, the fecal water content in electroacupuncture group significantly decreased(P<0.01); compared with the blank group, the fecal water content in model group and alosetron group was higher(P<0.001, P<0.05). Moreover, compared with the model group and alosetron group, the electroacupuncture group showed a decrease in fecal water content(P<0.01, P<0.05).(4) Intestinal volume threshold:Compared with the blank group, the intestinal volume threshold in model group significantly decreased(P<0.001). Besides, compared with the model group, the intestinal volume threshold of alosetron group and electroacupuncture group increased(P<0.05).(5) Tissue HE staining:Compared with the model group, the mucosal epithelium in alosetron group was intact, and the necrotic area was obviously reduced. Additionally, in the electroacupuncture group, the mucosa epithelium was also intact, the absorptive cells were columnar, and the inflammatory cells such as lymphocytes and a small amount of neutrophils were infiltrated in the mucosal folds. Compared with the alosetron group, the electroacupuncture group exhibited intact epithelium with a mild degree of necrotic inflammatory damage.(6) Pathway of 5-HT determination:Compared with the blank group, the serum 5-HT content in the model group increased(P<0.05), while compared with the model group, the serum 5-HT content in electroacupuncture group decreased(P<0.05).(7) Protein expression of colonic 5-HT3AR:Compared with the blank group, the model group showed an increased protein expression of colonic 5-HT3AR(P<0.05). In addition, compared with the model group, both the alosetron and electroacupuncture groups exhibited significantly decreased protein expression of colonic 5-HT3AR(P<0.001). Furthermore, compared with the alosetron group, the electroacupuncture group showed a significant reduction in colonic 5-HT3AR protein expression(P<0.001).Conclusion Under the guidance of "combining he and mu points from the same type of meridians of different fu-organs", electroacupuncture at "Tianshu"(ST25) and "Zusanli"(ST36) had notable therapeutic effects on IBS-D rats. Its mechanism of action may involve reducing the content of 5-HT in serum and the protein expression of 5-HT3AR in colon, restoring intestinal motility and visceral sensory function, thereby alleviating the clinical symptoms of IBS-D. |
| Key words: diarrheal irritable bowel syndrome combining he and mu points from the same type of meridians of different fu-organs visceral hypersensitivity 5-hydroxytryptamine 5-hydroxytryptamine 3A receptor |
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