温和灸对IBS小鼠结肠Trp-kyn/5-HT代谢的调控机制研究

Research on the regulatory mechanism of mild warm moxibustion on Trp-kyn/5-HT metabolism in the colon of IBS mice

目的 观察温和灸干预对肠易激综合征模型小鼠结肠组织Trp-Kyn/5-HT限速酶表达及相关代谢物含量的影响,阐释艾灸治疗相关机制。方法 雄性清洁级C57BL/6J小鼠随机分为正常组、模型组、艾灸组、1-MT组、PCPA组,采用AWR评价小鼠内脏高敏感状态,采用LC-MS/MS法、免疫组织化学法及Real Time PCR法分别检测小鼠结肠组织Trp-Kyn/5-HT代谢物含量、Trp-Kyn/5-HT代谢限速酶IDO1、TPH1蛋白及m RNA表达。结果 与正常组相比,模型组小鼠30 mm Hg下AWR评分增高(P <0.05),结肠组织TPH1、IDO1蛋白及m RNA表达显著增高(P <0.05),5-HT含量及Kyn/Trp比值显著升高(P <0.05)。与模型组相比,艾灸组和1-MT组小鼠结肠组织IDO1蛋白表达明显下调(P <0.05),艾灸组和PCPA组小鼠结肠组织TPH1蛋白明显下调(P <0.05);1-MT组小鼠结肠组织IDO1 mRNA表达明显降低(P <0.05),PCPA组小鼠结肠组织TPH1 mRNA表达明显降低(P <0.05);艾灸组Kyn/Trp比值显著降低(P <0.05)。结论 温和灸可抑制IBS小鼠结肠过表达的IDO1及THP1蛋白活性,下调Kyn/Trp比值,可能是温和灸改善IBS模型小鼠内脏痛敏的机制之一。

Objective To observe the effects of mild warm moxibustion intervention on the expression of Trp-Kyn/5-HT rate-limiting enzyme and the content of related metabolites in the colon tissue of irritable bowel syndrome(IBS)mice model,and to elucidate the mechanisms of the moxa-wool moxibustion treatment.Methods Male clean C57BL/6J mice were randomly divided into a normal group,a model group,a moxibustion group,a 1-MT group and a PCPA group.The visceral hypersensitivity of mice was assessed by AWR,and the content of Trp-Kyn/5-HT metabolites,the protein and mRNA expressions of IDO1 and TPH1 in the colon tissue of mice were detected by LC-MS/MS,immunohistochemistry,and Real Time PCR,respectively.Results Compared with the normal group,the AWR score of the model group was increased under 30 mmHg(P<0.05),and the protein and mRNA expressions of TPH1,IDO1 in the colon tissue were significantly increased(P<0.05),the content of 5-HT and the Kyn/Trp ratio were significantly increased(P<0.05).Compared with the model group,the protein expression of IDO1 in the colon tissue of mice in the moxibustion group and 1-MT group was significantly down-regulated(P<0.05),while the TPH1 protein in the colon tissue of mice in the moxibustion group and PCPA group was significantly down-regulated(P<0.05),and the mRNA expression of IDO1 in the colon tissue of mice in the 1-MT group was significantly reduced(P<0.05),while the mRNA expression of TPH1 in the colon tissue of mice in the PCPA group was significantly decreased(P<0.05),and the Kyn/Trp ratio was significantly decreased in the moxibustion group(P<0.05).Conclusion Mild warm moxibustion can inhibit the overexpression of IDO1 and THP1 protein activity in the colon tissue of IBS model mice,down-regulate the Kyn/Trp ratio,which may be one of the mechanisms of the treatment on improving visceral hyperalgesia in IBS model mice.