“标本配穴”针刺法对腹泻型肠易激综合征焦虑模型大鼠焦虑样行为及肠道菌群的影响

Effect of Biaoben(标本)Acupoints Combination Needling on Anxiety-like behavior and Gut flora in Model Rats of Irritable Bowel Syndrome with Predominant Diarrhea and Anxiety

目的探讨“标本配穴”针刺法治疗腹泻型肠易激综合征(IBS-D)伴焦虑的可能作用机制。方法从60只SD大鼠中随机抽取10只作空白组,剩余50只大鼠制备IBS-D焦虑模型,将造模成功的30只大鼠随机分为模型组、常规取穴组、标本配穴组,每组10只。模型组不干预,常规取穴组予“天枢”“足三里”“上巨虚”针刺干预,标本配穴组予“内关”“足三里”“关元”针刺干预,两组均每次针刺20 min,每天1次,共干预14天。各组大鼠进行Bristol粪便性状量表评分及腹壁撤退反射实验,并进行糖水偏好实验、旷场实验、高架十字迷宫实验评价其焦虑状态,绝对荧光定量PCR法检测肠道大肠杆菌、肠球菌、拟杆菌、乳酸杆菌、双歧杆菌、消化链球菌、芽孢杆菌7种目的菌的含量。结果与空白组比较,模型组大鼠Bristol粪便性状量表评分升高,糖水偏好率降低,大鼠在扩张压力为40、60、80 mmHg时腹壁撤退反射评分均升高,内脏疼痛阈值降低,旷场试验大鼠运动距离、直立次数、中央区域停留时间均减少,高架十字迷宫实验大鼠开放臂停留时间、进入开放臂次数、总路程均显著减少(P<0.05)。与模型组比较,常规取穴组和标本配穴组上述各指标均显著改善(P<0.05),且标本配穴组在改善糖水偏好率、内脏疼痛阈值及旷场试验、高架十字迷宫实验指标方面优于常规取穴组(P<0.05)。肠道菌群方面,常规取穴组、标本配穴组均可降低大肠杆菌、肠球菌、拟杆菌、消化链球菌的含量,升高乳酸杆菌、双歧杆菌、芽孢杆菌的含量,且以标本配穴组效果更明显(P<0.05)。结论标本配穴针刺法能显著改善IBS-D焦虑模型大鼠粪便性状和焦虑样负性情绪,其机制可能与调控肠道有益菌和致病菌菌群含量有关。

Objective To explore the possible mechanism of Biaoben(标本)acupoints combination needling in the treatment of irritable bowel syndrome with predominant diarrhea(IBS-D)with anxiety.Methods Ten rats were randomly selected from 60 SD rats as blank group and the remaining rats were prepared with the IBS-D and anxiety model.The successful modeling rats were randomly divided into model group,routine acupoints needling group and Biaoben acupoints combination needling group,with 10 rats in each group.The model group was left untreated.In the routine acupoints needling group,“Tianshu(ST25)”,“Zusanli(ST36)”and“Shangjuxu(ST37)”were selected,while in the Biaoben acupoints combination needling group,“Neiguan(PC6)”,“Zusanli(ST36)”,and“Guanyuan(CV4)”were used.Both groups received acupuncture for 20 minutes each time,once a day for 14 days.Bristol stool form scale(BSFS)and abdominal withdrawal reflex were assessed.Anxiety was evaluated by sucrose preference test,open field test,and elevated plus maze test.The absolute fluorescence quantitative PCR method was used to detect Escherichia coli,Enterococcus,Bacteroides,Lactobacillus,Bifidobacterium,Peptostreptococcus,and Bacillus contents.Results Compared to those in the blank group,the model group had higher BSFS score,lower sucrose preference percentage,increased abdominal withdrawal reflex score at the diastolic pressure of 40,60,and 80 mmHg,decreased visceral pain threshold,reduced distance of locomotion,number of uprights,and dwell time in the central area in the open field test,as well as the dwell time in the open arm,number of times into the open arm,and total distance in the elevated plus maze(P<0.05).Compared to those in the model group,the above-mentioned outcomes were significantly improved in both needling groups(P<0.05),and more improvements were found in the Biaoben acupoints combination needling group in terms of the sucrose preference percentage,visceral pain threshold,open field test and elevated plus maze indicators(P<0.05).Regarding the gut flora,both needling groups could decrease the content of Escherichia coli,Enterococcus,Bacteroides,and Peptostreptococcus,and increase the content of Lactobacillus,Bifidobacterium,and Bacillus,with more improvements in the Biaoben acupoints combination needling group(P<0.05).Conclusion The Biaoben acupoints combination needling is effective in improving the stool properties and relieving the anxiety-like negative emotions of IBS-D rats with anxiety,and the mechanism may be related to the regulation of the number of intestinal beneficial and pathogenic bacteria.