摘要
目的 采用代谢组学技术,分析识别结肠组织的代谢模式,探讨针灸抑制腹泻型肠易激综合征(irritable bowel syndrome with predominant diarrhea, IBS-D)模型大鼠肠道炎症反应的作用机制。方法 随机将45只SD大鼠分为3组,即对照组、模型组、针灸组。模型组与针灸组采用急、慢性应激相结合方法制备IBS-D模型大鼠。针灸组在造模同时给予针灸“足三里”“关元”“内关”治疗,每日1次,共28日。通过观察稀便率、结肠组织病理学变化、ELISA检测血清和结肠黏膜肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白介素(interleukin, IL)-6、IL-1β含量及测定结肠转运功能评价肠道功能。利用TM广靶代谢组学技术对三组大鼠的结肠组织样品进行代谢轮廓分析,利用PCA和OPLS-DA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找到富集代谢通路。结果 造模后,模型组稀便率较对照组升高,差异有统计学意义(P<0.01)。与模型组比较,针灸组稀便率降低(P<0.01)。模型组与对照组比较血清TNF-α、IL-6、IL-1β含量上升,差异有统计学意义(P<0.05);针灸组与模型组比较血清TNF-α、IL-6、IL-1β含量下降,差异有统计学意义(P<0.05)。模型组与对照组比较结肠黏膜TNF-α、IL-1β含量上升,差异有统计学意义(P<0.05)。模型组与对照组比较,结肠黏膜IL-6含量差异无统计学意义(P>0.05)。针灸组与模型组比较,结肠黏膜TNF-α、IL-6、IL-1β含量差异均无统计学意义(P>0.05)。与对照组比较,模型组玻璃小球排出时间减少(P<0.05);与模型组比较,针灸组玻璃小球排出时间增加(P<0.05)。模型组大鼠结肠组织可见明显粘膜层厚度变薄,较多腺体消失,局部可见明显腺体损伤,同时可见较多细胞核固缩胞体消失,空洞明显;针灸组大鼠有较多炎症细胞浸润于粘膜层顶端,少量腺体损伤。与对照组比较,模型组的代谢物肉碱C13:0上调,肉碱C7:DC水平下调,L-苯丙氨酸-L-亮氨酸水平上调,环(酪氨酸—亮氨酸)水平上调,甘油2-磷酸水平下调;与模型组比较,针灸组代谢物肉碱C13:0水平下调,肉碱C7:DC水平上调,L-苯丙氨酸-L-亮氨酸水平下调,环(酪氨酸-亮氨酸)水平下调,甘油2-磷酸水平上调。模型组与对照组对比差异代谢途径有2条,分别是色氨酸代谢、类固醇激素生物合成。针灸组与模型组对比差异代谢通路有1条,是卟啉与叶绿素代谢。结论 从结肠组织代谢组学的角度,针灸可能是通过调节代谢物肉碱C13:0、肉碱C7:DC、L-苯丙氨酸-L-亮氨酸、环(酪氨酸—亮氨酸)、甘油2-磷酸的水平,调节卟啉与叶绿素代谢通路,发挥抑制肠易激综合征肠道炎症反应的作用。
Objective Metabolomics technique was used to analyze and identify the metabolic pattern of colon tissue,and to explore the mechanism by which acupuncture and moxibustion inhibit intestinal inflammatory reaction in IBS-D model rats.Methods Forty-five SD rats were randomly divided into three groups:a control group,a model group and an acupuncture and moxibustion group.In addition to the control group,the model group and acupuncture and moxibustion group used the combination of acute and chronic stress to prepare IBS-D model rats.In the acupuncture and moxibustion group,the acupoints of ST36,CV4 and PC6 were given to acupuncture and moxibustion once a day for^(2)8 days while modeling.Detection of TNF-α,IL-6,IL-1βin serum and colon mucosa by ELISA,and observing the rate of loose stool,histopathological changes of colon and the colonic transport function was measured to evaluate the intestinal function.The metabolic profile of colonic tissue samples of three groups of rats was analyzed by TM wide-target metabolomics technology,and multiple statistical analysis methods such as PCA and OPLS-DA were used,and the differential metabolites were screened by combining with the database,and the enriched metabolic pathway was found by KEGG.Results After modeling,the loose stool rate in the model group was significantly higher than that in the control group(P<0.01).Compared with the model group,the rate of loose stool in the acupuncture and moxibustion group was lower(P<0.01).Comparison of TNF-α,IL-6,IL-1βin serum between model group and control group,the difference was statistically significant(P<0.05).Comparison of TNF-α,IL-6,IL-1βin serum between acupuncture and moxibustion group and model group,the difference was statistically significant(P<0.05).Comparison of TNF-α,IL-6,IL-1βin colon mucosa between model group and control group,the difference was statistically significant(P<0.05).There was no statistically significant difference in IL-6 content in the colon mucosa between the model group and the control group(P>0.05).Comparison of TNF-α,IL-6,IL-1βin colon mucosa between acupuncture and moxibustion group and model group,the difference was no statistically significant(P>0.05).Compared with the control group,the ejection time of the model group decreased(P<0.05).Compared with the model group,the excretion time of glass globules in the acupuncture and moxibustion group increased(P<0.05).In the model group,significant thinning of the mucosal layer was observed in the colon tissue with more glands disappearing and obvious glandular damage visible locally,and more pyknotic cells disappeared and cavities were evident.In the acupuncture and moxibustion group,more inflammatory cells infiltrated into the top of the mucous membrane,and a small amount of glands were injured.Compared with the control group,the level of the metabolite Carnitine C13:0 in the model group was up-regulated,the level of Carnitine C7:DC was down regulated,the level of Phe-Leu was up regulated,the level of Cyclo(Tyr-Leu)was up regulated,and the level of Glucerol2-phosphate was down regulated;Compared with the model group,the level of the metabolite Carnitine C13:0 in the acupuncture and moxibustion group was down regulated,the level of Carnitine C7:DC was up regulated,the level of Phe-Leu was down regulated,the level of Cyclo(Tyr-Leu)was down regulated,and the level of Glucerol2-phosphate was up regulated.Compared with the control group,the model group had two different metabolic pathways,namely tryptophan metabolism and steroid hormone biosynthesis.Compared with the model group,the acupuncture and moxibustion group has one differential metabolic pathway,which is porphyrin and chlorophyll metabolism.Conclusion From the perspective of colonic metabolomics,acupuncture and moxibustion may play a role in inhibiting the intestinal inflammatory response of irritable bowel syndrome by regulating the levels of metabolites carnitine C13:0,carnitine C7:DC,L-phenylalanine L-leucine,Cyclo(Tyr-Leu),and Glycerol 2-phosphate.
作者
邱宇驰
韩红伟
张家玮
付伊萌
QIU Yuchi;HAN Hongwei;ZHANG Jiawei;FU Yimeng(Maternal and child health hospital of Hubei Province,Wuhan 430075,China)
出处
《环球中医药》
CAS
2024年第3期384-392,共9页
Global Traditional Chinese Medicine
基金
2022年度湖北省教育厅科学研究计划指导性项目(B2022114)。
关键词
针灸
肠易激综合征
结肠
代谢组学
acupuncture and moxibustion
irritable bowel syndrome
colon
metabolomics
