莫诺苷对晚期糖基化终末产物诱导人脐静脉内皮细胞损伤的代谢组学研究  

Metabolomic study of morroniside on human umbilical vein endothelial cells injury induced by advanced glycation end products

作　　者：刘俊超[1,2,3] 刘丹[1,2,3] 贾银芝 吴云[1,2,3] 王振中[1,2,3] 萧伟[1,2,3] L LIU Dan JIA Yin-zhi WU Yun WANG Zhen-zhong XIAO Wei(Jiangsu Kanion Pharmaceutical Co., Ltd., Lianyungang 222001, China State Key Laboratory of Pharmaceutical Process New-teach for Chinese Medicine, Lianyungang 222001, China The Key Laboratory for the New Technique Research of TCM Extraction and Purification, Lianyungang 222001, China)

机构地区：[1]江苏康缘药业股份有限公司,江苏连云港222001 [2]中药制药过程新技术国家重点实验室,江苏连云港222001 [3]中药提取精制新技术重点研究室,江苏连云港222001

出　　处：《中草药》2017年第13期2674-2681,共8页Chinese Traditional and Herbal Drugs

基　　金：科学技术部国家重大新药创制项目(2013ZX09402203)

摘　　要：目的研究晚期糖基化终末产物(AGEs)诱导损伤人脐静脉内皮细胞(HUVECs)模型导致的细胞内源性代谢物的变化及山茱萸环烯醚萜苷类特征成分莫诺苷对异常代谢的调节作用,寻找莫诺苷保护损伤HUVECs潜在的代谢生物标志物,探索莫诺苷保护AGEs诱导损伤HUVECs的作用机制。方法体外培养HUVECs,AGEs诱导制备HUVECs损伤模型。将细胞分为对照组、模型组和给药组,通过高效液相色谱-串联四极杆飞行时间质谱(UPLC/Q-TOF-MS)技术,对3组细胞样本进行分离测定。采用主成分分析(PCA)、偏最小二乘法-判别分析(PLS-DA)、正交偏最小二乘法-判别分析(OPLS-DA)等模式识别方法对变量数据进行分析,t检验进行显著性统计分析,筛选出潜在生物标志物。通过独特通路分析(IPA),构建代谢组学特征网络图。结果模式识别能够较好地区分对照组、模型组和给药组细胞;进一步根据VIP数值大小筛选出30个对分类有显著贡献的离子,定性鉴定出10种潜在代谢生物标志物,这些内源性物质在给药组细胞内的水平均得到了不同程度的恢复。找到5条代谢通路,构建了莫诺苷保护AGEs诱导HUVECs损伤的代谢组学特征网络图。结论莫诺苷可使AGEs诱导损伤的HUVECs中代谢物水平有不同程度的恢复,其保护作用可能与5条相关代谢通路的调节有关。Objective To research endogenous metabolites changes of model group on human umbilical vein endothelial cells（HUVECs）injury induced by advanced glycation end products（AGEs）and the accommodation mechanism of morroniside（active components in Cornus officinalis）on the abnormal metabolism.To find potential biomarkers which morroniside protected the injured HUVECs,and to explore mechanisms of morroniside in treatment of HUVECs injury induced by AGEs.Methods HUVECs were cultivated in vitro,HUVECs injury model was established by the induction of AGEs.Cells were divided into three groups,control group,model group,and treatment group.Cell samples of three groups were determined with UPLC-Q-TOF/MS.Pattern recognition methods including PCA,PLS-DA,and OPLS-DA were applied to analyze multivariate data,and t-test was used in significant statistical analysis.It was used to find out potential biomarkers.Metabolomic feature network graphs were constructed ingenuity pathway analysis（IPA）.Results In pattern recognition,control group,model group,and treatment group could be distinguished better from each other.According to VIP values,30 ions which had a significant contribution to the classification were screened further,10potential metabolic biomarkers were identified qualitatively.These endogenous substances of the cells in treatment group in vivo had varying degrees of recovery.Five metabolic pathways were identified,and a metabolomic feature network of morroniside to protect against HUVECs injury induced by AGEs was constructed.Conclusion Changed metabolities on HUVECs injury induced by AGEs can be certainly recovered by morroniside,and the treatment effect of morroniside can be connected with the regulation of five related metabolic pathways.

关 键 词：莫诺苷 晚期糖基化终末产物 人脐静脉内皮细胞 代谢组学 模式识别 独特通路分析 

分 类 号：R285.5[医药卫生—中药学]