基于UPLC/Q-TOF-MS技术的大黄牡丹汤治疗IBD大鼠的血清代谢组学研究  

Serum Metabolomic Study on Dahuang Mudan Decoction in Treatment of IBD Rats Based on UPLC/QTOF-MS

作　　者：农菲菲 罗爽[1] 赵钟祥[1] 罗霞[1] 林铭军 文彬[2] 周联[1] NONG Feifei;LUO Shuang;ZHAO Zhongxiang;LUO Xia;LIN Mingjun;WEN Bin;ZHOU Lian(School of Pharmaceutical Science,Guangzhou University of Chinese Medicine,Guangzhou 510006 Guangdong,China;Pi-Wei Institute,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China)

机构地区：[1]广州中医药大学中药学院,广东广州510006 [2]广州中医药大学脾胃研究所,广东广州510405

出　　处：《中药新药与临床药理》2019年第5期571-579,共9页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(81673668);广东省自然科学基金项目(2017A030313657)

摘　　要：目的从代谢组学角度探讨大黄牡丹汤治疗炎症性肠病(IBD)的作用机制。方法采用灌肠给予2,4,6-三硝基苯磺酸(TNBS)-乙醇(90mg·kg^-1)诱导IBD模型;SD大鼠随机分正常组、模型组、美沙拉嗪组(430mg·kg^-1)和大黄牡丹汤组(7.5g·kg^-1),每组8只,连续灌胃给药7d,每天1次。观察大鼠一般情况,测定体质量及疾病活动指数(DAI);全自动血液分析仪检测大鼠红细胞、白细胞、粒细胞和淋巴细胞数量;采用ELISA法检测血清中1L-6、TNF-α的含量;HE染色观察结肠组织病理变化。采用超高效液相色谱-串联四极杆飞行时间质谱(UPLC/Q-TOF-MS)技术建立大鼠血清代谢轮廓图,分析各组大鼠血清内源性代谢物的变化,鉴定潜在的生物标志物并进行生物学意义分析。结果与正常组比较,模型组大鼠的DAI评分明显升高(P<0.001),体质量明显降低(P<0.001),白细胞、红细胞、淋巴细胞及粒细胞数目均显著上升(P<0.05,P<0.01),血清中IL-6、TNF-α含量显著升高(P<0.01,P<0.001),结肠组织出现不同程度的水肿、糜烂,腺体破坏严重且紊乱,有隐窝炎及脓肿形成。与模型组比较,给药组大鼠的DAI评分明显降低(P<0.05,P<0.01,P<0.001),体质量明显升高(P<0.05,P<0.01),白细胞、红细胞、淋巴细胞和粒细胞数目均显著下降(P<0.05,P<0.01),血清中IL-6、TNF-α的含量显著降低(P<0.01),结肠黏膜上皮逐渐恢复,肠绒毛形态结构趋于完整,肠腺结构明显。结果鉴定出IBD大鼠血清中的8个生物标志物,大黄牡丹汤可以上调苯乙酰甘氨酸、鞘氨醇、植物鞘氨醇、黄嘌吟、色氨酸、吲哚的含量(P<0.05,P<0.01),下调甘氨酸、苯丙氨酸的含量(P<0.05,P<0.01)。结论大黄牡丹汤对TNBS-乙醇诱导的IBD大鼠具明显的治疗作用,可能与其改善血清中内源性代谢物水平,恢复体内正常代谢活动有关。Objective To investigate the mechanism of Dahuang Mudan decoction (DMD) on the treatment of inflammatory bowel disease (IBD) by metabolomics approach.Methods The IBD model was induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS)/ethanol (90 mg·kg^-1).SD rats were randomly divided into control group,model group,mesalazine group (430 mg·kg^-1) and DMD group (7.5 g·kg^-1),8 rats in each group,and were continuously given correspondent drugs by gavage once a day for 7 days.The general conditions of the rats were evaluated by measuring the body weight and disease activity index (DAI).The white blood cell counts (WBC),lymphocyte counts (Lymph),granulocyte counts(Gran) and red blood cell(RBC) counts were measured by automatic blood analyzer.The contents of inflammatory cytokines including IL-6 and TNF-α in serum were determined by ELISA.Pathological changes of colon tissues were observed by HE staining.Ultrahigh performance liquid chromatography-tandem quadrupole time-of flight mass spectrometry (UPLC/Q-TOF-MS) was used to establish the serum metabolic profile of rats,and analyze the changes of serum endogenous metabolites to identify the potential biomarkers.The biological significance of the treatments was then interpreted.Results Compared with the control group,DAI score of model group increased significantly (P < 0.001),and the body weight significantly decreased (P < 0.001).The counts of WBC,RBC,Lymph and Gran of model group rats were significantly increased (P < 0.05,P < 0.01).Meanwhile,IL-6,TNF-α contents of model group rats increased significantly (P < 0.01,P < 0.001).Different degree of edema,erosion,destroyed and disordered glands,fossae inflammation and abscess formation appeared in colon tissues of model group rats.Compared with model group,DAI scores of DMD and mesalazine groups rats significantly decreased (P < 0.05,P < 0.01,P < 0.001),and body weights increased significantly (P < 0.05,P < 0.01);WBC,RBC,Lymph and Gran counts were significantly decreased (P < 0.05,P < 0.01);and the contents of IL-6 and

关 键 词：大黄牡丹汤 炎症性肠病 血清代谢组学 生物标志物 超高效液相色谱-串联四极杆飞行时间质谱 

分 类 号：R285.5[医药卫生—中药学]