Study on the efficacy and mechanism of Umbilicus acupuncture on diarrhea-type irritable bowel syndrome based on gut microbiota and tryptophan metabolism
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摘要: 目的 探讨脐针通过调节肠道菌群及色氨酸代谢在腹泻型肠易激综合征(diarrhea-type irritable bowel syndrome，IBS-D)治疗上的作用机制。方法 选择30例符合罗马Ⅳ标准的IBS-D患者入组，予脐针治疗，疗程4周。征集30例健康志愿者作基线检测对照。观察健康志愿者及IBS组患者治疗前后的临床症状评分，检测粪便肠道菌群，检测血清色氨酸(TRP)、5-羟色胺(5-HT)、犬尿氨酸(KYN)及吲哚2，3-双加氧酶1(IDO1)浓度，以及结肠色氨酸羟化酶1(TPH1)蛋白表达。结果 经脐针治疗后，IBS患者临床症状评分较治疗前降低(P< 0.01)；肠道菌群多样性增加，结构差异明显；IBS患者血清TRP、5-HT、KYN、IDO1浓度较前下降(P< 0.01)；结肠TPH1蛋白表达上升(P< 0.05)。结论 脐针治疗能改善IBS-D患者临床症状，其作用机制可能与通过调节肠道菌群及色氨酸代谢，抑制5-HT及KYN生成相关。Abstract: Objective To investigate the mechanism of Umbilicus acupuncture on diarrhea-type irritable bowel syndrome(IBS-D) based on gut microbiota and tryptophan metabolism.Methods Thirty patients with IBS-D who met the Rome Ⅳ diagnostic criteria for IBS-D were enrolled and treated with Umbilicus acupuncture for 4 weeks. Thirty healthy volunteers served as baseline controls. The changes of clinical symptom scores before and after treatment were observed. Gut microbiota were also detected. And serum tryptophan(TRP), 5-hydroxytryptamine(5-HT), kynurenine(KYN), as well as indoleamine 2, 3-dioxygenase1(IDO1) were recorded. The colonic tryptophan hydroxylase1(TPH1) protein expression was detected before and after treatment.Results The clinical symptom scores of IBS patients after treatment in IBS group were lower than those before treatment(P< 0.01). The diversity of gut microbiota was increased, and significant differences were in gut microbiota structure. The levels of TRP, 5-HT, KYN and IDO1 were also decreased(P< 0.01). Colonic TPH1 expression levels were increased after treatment(P< 0.05).Conclusion Umbilicus acupuncture can improve the clinical symptoms of IBS-D patients, and its mechanism of action may be related to the inhibition of 5-HT and KYN production by regulating gut microbiota and tryptophan metabolism.
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表 1 2组患者治疗前后IBS-SSS评分
分，X±S 组别 例数 观测时点 IBS-SSS 正常组 30 20.17±4.11 IBS组 30 治疗前 281.80±14.371) 治疗后 102.70±10.112) 与正常组比较，1)P < 0.01；与IBS组治疗前比较，2)P < 0.01。 
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表 2 2组患者治疗前后Bristol评分及排便频率评分
分，M(P25，P75) 组别 例数 观测时点 Bristol评分 排便频率 正常组 30 4(4，5) 3(3，3) IBS组 30 治疗前 6(6，7)1) 5(4.75，6)1) 治疗后 4(4，5)2) 3(3，4)2) 与正常组比较，1)P < 0.01；与IBS组治疗前比较，2)P < 0.01。
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表 3 正常组和IBS组治疗前门及属水平差异菌群
菌群结构 正常组(丰度) IBS组治疗前(丰度) P 增减 门 Bacteroidota 0.313201197 0.250301947 0.037 减少 Cyanobacteria 0.000003460 0.001300590 ＜0.001 增加 Acidobacteriota 0.000001390 0.000043600 0.003 增加 属 Faecalibacterium 0.160179645 0.116923599 0.040 减少 Clostridium 0.002557550 0.006413613 0.046 增加 Fusicatenibacter 0.009211474 0.014392365 0.020 增加 Streptococcus 0.004804842 0.009632538 0.038 增加 Romboutsia 0.014589739 0.008054240 0.009 减少 Alistipes 0.018322530 0.007267514 0.001 减少
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表 4 IBS组治疗前后门及属水平差异菌群
菌群结构 IBS组治疗前(丰度) IBS组治疗后(丰度) P 增减 门 unidentified_Bacteria 0.001425247 0.002543699 0.040 增加 属 Faecalibacterium 0.116923599 0.151519433 0.017 增加 Klebsiella 0.005671210 0.001345605 0.006 减少 Agathobacter 0.022396050 0.033899831 0.005 增加 Clostridium 0.006413613 0.002336630 0.016 减少 Alloprevotella 0.002608798 0.000909305 0.038 减少 Alistipes 0.007267514 0.011891604 0.021 增加 Christensenellaceae 0.001611540 0.004626167 0.004 增加
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表 5 正常组及IBS组患者治疗前后TRP、5-HT、KYN、IDO1浓度
X±S 组别 例数 观测时点 TRP/(μmol·L-1) 5-HT/(ng·mL-1) KYN/(nmol·L-1) IDO1/(IU·mL-1) 正常组 30 51.45±1.77 164.61±7.98 1530.80±38.29 18.17±0.82 IBS组 30 治疗前 61.90±2.031) 265.48±9.691) 1795.14±36.791) 35.46±1.711) 治疗后 54.23±1.642) 219.17±8.772) 1587.12±33.732) 29.77±1.412) 与正常组比较，1)P < 0.01；与IBS组治疗前比较，2)P < 0.01。
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