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目的:探讨电针内关、足三里对IBS模型大鼠行为学及结肠5-HT2A受体表达的影响,从靶器官痛相关物质变化角度揭示不同穴位干预慢性内脏痛的部分分子机制。方法:采用新生期母婴分离和醋酸灌肠并在后期结直肠球囊扩张刺激法联合制备肠易激综合征大鼠模型。分为空白对照组、模型组、内关穴组、足三里穴组。空白对照组和模型组不做任何处理,内关穴组和足三里穴组进行电针干预。采用腹部回撤反射及高架十字迷宫实验评价IBS模型大鼠的肠道敏感性和情绪心理变化及针刺不同穴位的干预效应。采用免疫组织化学方法检测大鼠结肠组织5-HT2A受体的免疫反应物的表达。结果:(1)腹部回撤反射:与空白对照组比较,模型组大鼠腹部回撤反射潜伏期缩短(P<0.01);模型组、内关穴组、足三里穴组的腹部回撤收缩波个数增加(P<0.01或P<0.05)。与模型组比较,内关穴组、足三里穴组的潜伏期延长(P<0.05或P<0.01),收缩波个数减少(P<0.05或P<0.01)。与内关穴组比较,足三里穴组的收缩波个数减少(P<0.05)。(2)高架十字迷宫:与空白对照组比较,模型组进入开放臂次数百分比减少(P<0.05),总距离减少(P<0.05),内关穴组开放臂停留时间百分比减少(P<0.01),总距离减少(P<0.05)。(3)5-HT2A受体表达:与空白对照组比较,模型组、内关穴组、足三里穴组的5-HT2A受体免疫反应物阳性的表达增强(P<0.01)。与模型组比较,足三里穴组的5-HT2A受体免疫反应物阳性表达减弱(P<0.01)。与内关穴组比较,足三里穴组5-HT2A受体免疫反应物阳性表达减弱(P<0.05)。结论:IBS模型大鼠内脏疼痛敏感性增高,情绪心理行为异常改变,符合IBS病理特征,表明母子分离加醋酸灌肠结合结直肠球囊扩张刺激可以成功制备IBS大鼠模型;针刺内关穴、足三里穴可降低IBS模型大鼠肠道敏感性,缓解内脏疼痛症状,且足三里穴优于内关穴。针刺可抑制5-HT2A受体免疫反应物的异常表达,参与疼痛的调节。
Objective: To investigate the effects of electro-acupuncture at Neiguan and Zusanli on behavior and colon 5-HT2A receptor expression in IBS model rats and to reveal some molecular mechanisms of different acupoint interventions on chronic visceral pain from the perspective of target organ-related substances. Methods: The model of irritable bowel syndrome (IBS) was established by neonatal maternal separation and acetic enema and colorectal balloon dilatation stimulation in the late stage. Divided into blank control group, model group, Neiguan point group, Zusanli point group. Blank control group and model group without any treatment, Neiguan acupuncture points and Zusanli acupoint intervention group. Abdominal retracement reflex and elevated plus maze test were used to evaluate the changes of intestinal sensitivity and emotion in IBS rats and the intervention effects of different acupuncture points. Immunohistochemistry was used to detect the expression of 5-HT2A receptor immunoreactive substance in rat colon tissue. Results: (1) Abdominal retraction reflex: compared with the blank control group, the abdomen retraction reflex latency of the model group was shortened (P <0.01); abdomen retraction retraction wave of the model group, Neiguan acupoint group and the Zusanli acupoint group The number increased (P <0.01 or P <0.05). Compared with the model group, the latency of Neiguan and Zusanli acupoints was prolonged (P <0.05 or P <0.01), and the number of contraction wave was decreased (P <0.05 or P <0.01). Compared with Neiguan acupoint group, the number of contraction wave in Zusanli acupoint group decreased (P <0.05). (2) Elevated plus maze: Compared with the blank control group, the percentage of open arms in the model group decreased (P <0.05), the total distance decreased (P <0.05) ), The total distance decreased (P <0.05). (3) The expression of 5-HT2A receptor: Compared with the blank control group, the expression of 5-HT2A receptor immunoreactivity in model group, Neiguan acupoint group and Zusanli acupoint group was enhanced (P <0.01). Compared with the model group, the positive expression of 5-HT2A receptor immunoassay in the Zusanli acupoint group was weakened (P <0.01). Compared with Neiguan acupoint group, the positive expression of 5-HT2A receptor immunoreactivity in Zusanli acupoint group was weakened (P <0.05). Conclusion: IBS model rats visceral pain sensitivity increased, changes in emotional and psychological behavior, in line with IBS pathological features, indicating that mother and child separated plus acetic acid enema with colorectal balloon dilatation stimulation can be successfully prepared IBS rat model; acupuncture at Neiguan, The Zusanli acupoint can reduce the intestinal sensitivity of IBS model rats, relieve the visceral pain symptoms, and the Zusanli points are better than the Neiguan points. Acupuncture can inhibit the abnormal expression of 5-HT2A receptor immune response, involved in the regulation of pain.