基于细胞代谢组学技术的人参皂苷Rb1对SH-SY5Y细胞保护机制研究  

Metabolomics Study of Protection Mechanism of Ginsenoside Rb_1 on Injury of SH-SY5Y Cell Induced by Glutamic Acid

作　　者：杨银平 律广富 张乔 姚梦杰 梁新合 李茎 张辉 孙佳明 YANG Yin-Ping;LV Guang-Fu;ZHANG Qiao;YAO Meng-Jie;LIANG Xin-He;LI Jing;ZHANG Hui;SUN Jia-Ming(Jilin Ginseng Academy,Changchun University of Chinese Medicine,Changchun 130117,China)

机构地区：[1]长春中医药大学吉林省人参科学研究院,长春130117

出　　处：《分析化学》2019年第1期49-58,共10页Chinese Journal of Analytical Chemistry

基　　金：国家自然基金项目(Nos.31570347;81704001);吉林省科技发展计划项目(No.20180414072GH);吉林省卫生与计划生育委员会科研计划项目(No.2018J111)资助~~

摘　　要：采用基于1H NMR的细胞代谢组学技术探讨人参皂苷Rb1对谷氨酸诱导SH-SY5Y细胞凋亡的保护作用机制。应用MTT比色法及流式细胞术分析SH-SY5Y细胞在造模和给药前后的细胞存活率和凋亡率,利用1H NMR技术平台结合多元统计分析对细胞代谢的动态变化进行研究,揭示SH-SY5Y细胞在造模和给药前后内源性代谢物变化情况。结果表明,人参皂苷Rb1能通过减少谷氨酸诱导SH-SY5Y细胞凋亡而产生保护作用,结合正交偏最小二乘法判别分析(OPLS-DA)共鉴定出谷氨酸、牛磺酸、乙酸、精氨酸、肌酸、丙氨酸和脯氨酸共7个差异代谢物。经Metaboanalyst分析,主要涉及牛磺酸和亚牛磺酸代谢、精氨酸和脯氨酸代谢、丙氨酸、天冬氨酸和谷氨酸代谢、D-谷氨酰胺和D-谷氨酸代谢、丙酮酸代谢共5条能量代谢及氨基酸代谢途径。结果表明,人参皂苷Rb1主要通过影响多种能量及氨基酸代谢对氧化损伤的神经细胞产生保护作用。Metabolomics based on 1H nuclear magnetic resonance(NMR)was established to explore the protection mechanism of ginsenoside Rb 1 on injury of SH-SY5Y cell induced by glutamic acid(Glu).Cell viability was evaluated by methyl thiazolyl tetrazolium(MTT)method,apoptotic rates were analyzed with PI/Annexin V flow cytometry,and the dynamic changes of cell metabolism were studied by using 1H NMR technology platform and high-throughput multivariate statistical analysis to reveal the changes of endogenous metabolites of SH-SY5Y cell.It was found that ginsenoside Rb 1 could protect the SH-SY5Y cell by reducing apoptosis induced by glutamate and 7 metabolites of glutamic acid,taurine,acetic acid,arginine,creatine alanine,and proline were identified through orthogonal partial least squares discriminant analysis(OPLS-DA).Metaboanalyst showed taurine and hypotaurine metabolism,arginine and proline metabolism,alanine,aspartate and glutamate metabolism,D-Glutamine and D-glutamate metabolism and pyruvate metabolism were mainly involved.The results showed that ginsenoside Rb 1 had a protective effect on SH-SY5Y cell by affecting various energy and amino acid metabolism.

关 键 词：人参皂苷RB1 细胞代谢组学 1H核磁共振 凋亡保护 正交偏最小二乘法判别分析 

分 类 号：R285[医药卫生—中药学]