i TRAQ联合NanoLC-LTQ-Orbitrap技术分析二仙汤干预去卵巢骨质疏松大鼠的骨组织差异表达蛋白  

Analysis of the Differentially Expressed Proteins of Ovariectomized Osteoporosis Rats with Er-xian Decoction Intervention by iTRAQ Coupled with NanoLC-LTQ-Orbitrap

作　　者：刘波[1,2] 刘志文 吴琪[1] 谢慧慧[1] 杨硕[1,2] 王莹[1] LIU Bo;LIU Zhiwen;WU Qi;XIE Huihui;YANG Shuo;WANG Ying(Pharmacy College, Jiangxi University of Traditional Chinese Medicine,Nanchang 330004 Jiangxi,China;Jiangxi Key laboratory of Chinese Medicine Pharmacology,Nanchang 330004 Jiangxi,China)

机构地区：[1]江西中医药大学药学院,江西南昌330004 [2]江西省中药药理学重点实验室,江西南昌330004

出　　处：《中药新药与临床药理》2019年第2期194-199,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(81860802;81560723);江西省自然科学基金项目(20151BAB205072)

摘　　要：目的筛选以二仙汤干预去卵巢骨质疏松大鼠后的差异表达蛋白质,探讨二仙汤防治骨质疏松的分子作用机制。方法雌性SD大鼠,按体质量随机分为3组,即假手术组、模型组、二仙汤组,其中假手术组和模型组灌胃生理盐水,二仙汤组按生药量12 g·kg^(-1)灌胃,每日1次,连续给药90 d。取大鼠股骨,运用同位素相对和绝对定量(Isobaric tags for relative and absolute quantitation,iTRAQ)联合纳升高效液相色谱-线性离子阱-静电场轨道阱(NanoLC-LTQ-Orbitrap)高分辨质谱技术的定量蛋白质组学方法检测骨蛋白,采用ProteinDiscovery软件进行分析和鉴定差异蛋白质,通过生物信息学分析差异表达的蛋白质功能。结果质谱分析共鉴定出862个差异蛋白,其中二仙汤组与模型组相比较,共发现82个与假手术组趋势一致的差异蛋白,上调蛋白45个,下调蛋白37个。生物信息学分析发现,差异表达的蛋白大多数属于水解酶、核酸结合类蛋白、受体类蛋白等。结论细胞色素b-245轻链、促分裂原活化蛋白激酶-1、Ras相关C3肉毒杆菌毒素底物-1等相关差异蛋白质可能是二仙汤防治骨质疏松症的作用靶点。Objective To detect protein expression differences after Er-xian decoction intervened the ovariectomized osteoporosis rats and to explore the molecular mechanism of Er- xian decoction on prevention and cure of the osteoporosis. Methods Female SD rats were randomly divided into three groups, the Sham group, control group, and Er-xian decoction group. Rats in Sham group and control group were gavaged with normal saline every day,and the rats in Er-xian decoction group were gavaged with the dose of 12 g·kg^-1 once a day. All the treatments were last for 90 d. At the end of the treatments,the bone proteins were detected by iTRAQ-coupled NanoLC-LTQ-Orbitrap, and identified by Protein Discovery software. Functions of identified proteins were analyzed by bioinformatics. Results A total of 862 differentially expressed proteins were identified. Compared with control group, 82 differentially expressed proteins had the variation trends consistent with the Sham group were found in the Er- xian decoction group;among which 45 were up- regulated, 37 were down- regulated. These differentially expressed proteins included hydrolase,nucleic acid binding and receptor protein,etc. Conclusion The differentially expressed proteins,such as Cytochrome b-245 light chain,Mitogen-activated protein kinase 1,Ras-related C3 botulinum toxin substrate 1 may be the targets of Er-xian decoction.

关 键 词：同位素相对和绝对定量 纳升高效液相色谱-线性离子阱-静电场轨道阱高分辨质谱 二仙汤 骨质疏松症 蛋白质组学 生物信息学 

分 类 号：R285.5[医药卫生—中药学] R857.3[医药卫生—中医学]