Objective: To investigate the effect of acupuncture-moxibustion with "Biao-Ben" acupoint combination (BB) on the serum metabolites and metabolic pathway in irritable bowel syndrome (IBS) model rats, so as to explore the mechanisms of BB in the prevention and treatment of IBS.
Methods: Thirty SD rats were randomly divided into three groups: control group, model group, and BB group, with 10 rats in each group. The IBS model was established by the combination of acute stress method and chronic stress method, and the success of the model establishment was evaluated by abdominal wall reflex (AWR). BB group received acupuncture-moxibustion treatment at "Neiguan" (PC6), "Zusanli" (ST36), and "Guanyuan" (CV4) for 15 min, once a day, for a total of 28 days. Bristol's fecal character score was evaluated, and intestinal propulsion rate was calculated. The open-field experiment was used to observe the behaviour of rats. Pathological changes in the colon were observed by H.E. staining. TM widely targeted metabolomics technology was used to detect the metabolic profile of serum samples from 3 groups of rats. Principal component analysis and orthogonal partial least-squares discrimination analysis techniques were combined with database screening to screen differential metabolites, and the KEGG database was utilized to map the enriched metabolic pathway.
Results: Compared with the control group, AWR, and the total distance, speed, duration traveled autonomously, the distance of central grid traveling, the number of central grid crossings of the open-field experiment were significantly decreased (P<0.01，P<0.05), while Bristol's fecal character score, intestinal propulsion rate and rest duration in the open-field experiment were significantly increased(P<0.01) in the model group. Compared with the model group, Bristol's fecal character score, the intestinal propulsion rate, rest duration, and rest episode were significantly decreased(P<0.01, P<0.05), while AWR, the total distance, speed, duration traveled autonomously, the distance of central grid traveling, the number of central grid crossings, and the residence time of the central grid were significantly increased(P<0.01，P<0.05) in the BB group. H.E. staining showed a discontinuous mucosal layer of colon tissue, a slightly disordered arrangement of glands, and more inflammatory cell infiltration in the submucosa and muscle layer in the model group, which was relatively milder in the BB group. After comparing the model and control group, 123 differential metabolites were screened, while 57 were screened after comparing the model and BB group. Six differential metabolic pathways were acquired when comparing the model and the control group, while 8 were acquired when comparing the model and BB group using KEGG enrichment analysis, both of which included the arachidonic acid metabolism pathway.
Conclusion: "Biao-Ben" acupoint combination can improve symptoms of IBS by regulating metabolites of the arachidonic acid metabolism pathway, which may be a potential target for the treatment of IBS.
目的：基于TM广靶代谢组学的技术，探讨标本配穴针灸对肠易激综合征(IBS)大鼠代谢模式的影响。方法：SD大鼠随机分为对照组、模型组和标本配穴组，每组10只。通过慢性、急性应激结合的方法制备IBS模型。标本配穴组给予针刺+艾灸“内关”“足三里”“关元”，每次15 min，1次/d，共28 d。测定腹壁撤退反射(AWR)压力阈值，评价Bristol粪便性状评分，计算小肠推进率，旷场实验对大鼠的行为学进行评定，HE染色法观察结肠的病理改变。利用TM广靶代谢组学技术对3组大鼠的血清样品进行代谢轮廓分析，利用主成分分析和正交偏最小二乘判别分析等多元统计分析方法，并结合数据库筛选差异代谢物，利用KEGG数据库进行代谢通路富集分析。结果：治疗后与对照组比较，模型组AWR压力阈值显著降低(P<0.01)，Bristol粪便性状评分和小肠推进率显著升高(P<0.01)。与模型组比较，标本配穴组AWR压力阈值显著升高(P<0.01)，Bristol粪便性状评分和小肠推进率显著降低(P<0.01，P<0.05)。旷场实验中，对照组大鼠的运动轨迹自由且总路程长，模型组大鼠的运动轨迹多在外围且总路程短；与对照组比较，模型组运动总距离、中央运动距离、速度、运动持续时间、穿格次数减少(P<0.01，P<0.05)，休息持续时间增加(P<0.01)；与模型组比较，标本配穴组运动总距离、速度、运动持续时间、中央运动距离、中央格停留时间、穿格次数增加(P<0.01)，休息片段、休息持续时间减少(P<0.05，P<0.01)。模型组大鼠结肠组织黏膜层内可见大量炎性细胞浸润，且肠腺厚度变薄，溃疡严重；标本配穴组大鼠结肠组织肠腺结构基本完整，杯状细胞数量较丰富，中度溃疡。与对照组比较，模型组共筛选出差异代谢物有123个；与模型组比较，标本配穴组共筛选出差异代谢物有57个。利用KEGG富集分析得出模型组与对照组有6条差异代谢通路，标本配穴组与模型组比较有8条差异代谢通路，均包含花生四烯酸代谢通路。结论：花生四烯酸代谢通路可能是标本配穴防治IBS的潜在生物标记物和潜在靶点。.
Keywords: Acupuncture and moxibustion; Irritable bowel syndrome; TM widely targeted metabolomics; “Biao-Ben” acupoint combination.