同位素标记相对和绝对定量技术在药物蛋白质组学研究中的应用

目的建立应用同位素标记相对和绝对定量(iTRAQ)技术进行药物蛋白质组学研究的技术平台。方法分别测定12例高血压患者在给予降压药氨氯地平治疗前和治疗后8周的收缩压和舒张压,并同时收集患者在降压药治疗前和治疗后8周的血清,应用蛋白质组学iTRAQ技术对治疗前后的血清蛋白质组进行定量分析。结果高血压患者口服降压药苯磺酸氨氯地平片8周后平均收缩压和舒张压均明显降低(P<0.01)。经iTRAQ和ProteinPilot软件分析,共鉴定出232个差异蛋白质,其中降压药物治疗后表达上调和下调超过1.5倍的蛋白质分别有12种和13种。上调的蛋白质主要参与应激、防御和免疫反应,而下调的蛋白质主要参与应激、防御、炎症和凝血反应。本研究结果提示,降压药氨氯地平治疗可以双向调节机体的应激和防御反应,可以增强机体的免疫力,抑制炎症反应和改善高血凝状态,从而有利于血压的降低。结论应用iTRAQ技术进行药物蛋白质组学研究可筛选获得多种与药物疗效相关的差异蛋白质,为药物作用靶点和分子机制等方面的研究提供了一个良好的技术平台。

Application of Isotope Tagging for Relative and Absolute Protein Quantitation in the Drug Proteomic Study

Objective To establish the technology platform of isotope tagging for relative and absolute protein quantitation（iTRAQ） for the drug proteomic study.Methods The systolic and diastolic blood pressure prior to therapy and 8 weeks after therapy with amlodipine were determined in 12 patients with hypertension,and at the same time,the individual serum from each patient was collected for iTRAQ analyses.Results The systolic and diastolic blood pressure were signifi cantly decreased 8 weeks after therapy with amlodipine compared with those prior to therapy in 12 patients with hypertension（P0.01）.Total 232 differentially expressed proteins were identif ied by iTRAQ and ProteinPilot software analyses,of which 12 proteins were up-regulated with above 1.5-fold and 13 proteins were down-regulated with above 1.5-fold 8 weeks after therapy.The up-regulated proteins mainly involved in stress,defense and immune response.Whereas,the down-regulated proteins mainly participate in stress,defense,in ammatory response,and blood coagulation.These results suggested that antihypertensive treatment with amlodipine exert biphasic regulation effects on stress and defense,enhance immunity,inhibit in ammatory response,and improve hypercoagulable state.Conclusion iTRAQ technology may help us to identify differentially expressed proteins in drug proteomic study,which is benef icial to clarifi cation of targets and molecular mechanism of drugs.