基于UPLC-MS/MS的热毒宁注射液治疗大鼠急性肺损伤的血浆代谢组学研究  

Metabonomic study on the anti-acute lung injury effect of Reduning injection based on UPLC-MS/MS in rat plasma

作　　者：汪衍敏 王艳娟[1] 赵龙山[1] 王振中[2] 萧伟[2] 熊志立[1] WANG Yanmin;WANG Yanjuan;ZHAO Longshan;WANG Zhenzhong;XIAO Wei;XIONG Zhili(School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China;Jiangsu Kanion Pharmaceutical Co. ,Ltd.,Lianyungang 222001, China)

机构地区：[1]沈阳药科大学药学院,辽宁沈阳110016 [2]江苏康缘药业股份有限公司,江苏连云港222001

出　　处：《沈阳药科大学学报》2018年第5期374-380,共7页Journal of Shenyang Pharmaceutical University

基　　金：国家自然科学基金资助项目(81403062)

摘　　要：目的采用基于超高效液相色谱串联质谱(UPLC-MS/MS)技术的代谢组学方法,分析大鼠血浆内源性代谢物的变化,研究热毒宁注射液治疗急性肺损伤的作用机制。方法将21只SD大鼠随机分为3组,即对照组、模型组和热毒宁治疗组。模型组和热毒宁治疗组通过尾静脉注射内毒素(lipopolysaccharide,LPS,7.5 mg·kg-1)建立急性肺损伤(acute lung injury,ALI)模型,治疗组0.5 h后注射热毒宁注射液(5 m L·kg-1)。6 h后,收集3组血浆样品,采用UPLC-MS/MS技术分析测定大鼠血浆代谢物谱,对得到的数据进行偏最小二乘法-判别分析(partial least squares discriminant analysis,PLS-DA)。结果对照组、模型组和热毒宁治疗组血浆代谢物谱明显分离,发现并鉴定了8个潜在的生物标志物。与对照组大鼠相比,模型组大鼠血浆中溶血磷脂酰胆碱LPC 18∶2、LPC 20∶4、LPC 16∶0、LPC 18∶0和肌酸的浓度显著降低,色氨酸、苯丙氨酸和不对称二甲基精氨酸(asymmetric dimethylarginine,ADMA)的浓度显著升高。给药后,大鼠体内的相关生物标志物水平趋于正常。结论这些发生显著变化的标志物与体内氨基酸代谢、能量代谢和磷脂类代谢有关,推测热毒宁注射液通过调节这些代谢途径来达到治疗急性肺损伤的作用。本实验同时表明,代谢组学可以作为一个研究平台来探究中药复方的整体效应及其作用机制。Objective A metabonomic method based on UPLC-MS/MS was established to study the plasma metabolic changes in rat acute lung injury（ ALI） model,and to investigate therapeutic effect of Reduning（ RDN） injection. Methods Twenty-one male Sprague-Dawley（ SD） rats were divided into three groups：control group,model group and treatment group. The model and treatment rats were challenged with intravenous injection of lipopolysaccharide（ LPS,7. 5 mg·kg-1） to induced ALI,and treatment group was given a single dose of RDN（ 5 m L·kg-1） 30 min after LPS challenged. After 6 h,all plasma samples of the three groups were collected and there metabolic profiling changes were analyzed by UPLC-MS/MS,and the resulting dataset was analyzed by partial least squares discriminant analysis（ PLS-DA）. Results Obvious separation trend was found among three groups,and eight potential biomarkers were identified in the rat plasma. Comparing with the control group,the levels of LPC 18 ∶ 2,LPC 20 ∶ 4,LPC 16 ∶ 0,LPC 18 ∶ 0 and creatine were decreased in model group,while the levels of tryptophan,phenylalanine and asymmetric dimethylarginine（ ADMA） were increased. The levels of these metabolites were recovered in different degrees after Reduning treatment. Conclusions Based on the identified biomarkers,we speculated that RDN could provide anti-ALI effect through regulating the perturbed metabolic pathways including amino acid metabolism,energy metabolism and phospholipid metabolism. This work also highlights that metabonomics is a potentially helpful tool for studying the holistic effects and action mechanism of TCM.

关 键 词：热毒宁注射液 急性肺损伤 代谢组学 UPLC-MS/MS 大鼠血浆 

分 类 号：R917[医药卫生—药物分析学]