基于iTRAQ技术结合2D-LC-MS/MS研究黄连对CYP450同工酶表达的影响  

iTRAQ technology combined with 2D-LC-MS/MS to analyze effect of Coptidis Rhizoma on cytochrome P450 isoenzyme expression

作　　者：杨欣[1] 王秋红[1] 王蒙[1] 王悦[1] 杨炳友[1] 匡海学[1] 

机构地区：[1]黑龙江中医药大学教育部北药基础与应用研究重点实验室黑龙江省中药及天然药物药效物质基础研究重点实验室,黑龙江哈尔滨150040

出　　处：《中国中药杂志》2016年第4期731-736,共6页China Journal of Chinese Materia Medica

基　　金：国家重点基础研究发展计划(973)项目(2013CB531801);黑龙江中医药大学研究生创新科研项目(2015009)

摘　　要：黄连是临床常用药材,在中医药中被广泛应用,具有清热燥湿、泻火解毒之功,中药成分比较复杂,细胞色素P450(cytochrome P450,CYP450)同工酶是药物的主要代谢酶,研究黄连对CYP450酶的影响具有很重要的临床意义。实验选用大鼠肝脏组织,差速离心法分离肝微粒体,BCA(bicinchoninic acid)蛋白定量,应用同位素标记相对和绝对定量(isobaric tags for relative and absolute quantification,iTRAQ)技术结合2D-LC-MS/MS筛选差异蛋白。黄连一共鉴定出30个CYP450同工酶,7个表达上调,8个表达下调,15个没有发生变化。iTRAQ技术结合2D-LC-MS/MS可以全面研究CYP450酶,但黄连有必要进一步在体外水平对其进行评价,预防潜在的临床药物相互作用。Coptidis Rhizoma is commonly used in clinical medicine. It has been widely used in traditional Chinese medicine（ TCM）,with the functions of clearing heat,drying dampness,purging intense heat relieve toxins. However,the herb contains complex ingredients,and CYP450 isoenzyme is the main metabolic enzyme of the drug,so it is of important clinical significance to study the effect of Coptidis Rhizoma on cytochrome P450 enzymes. In the experiment,liver tissues of rats were selected and liver microsomes were separated by differential centrifugation. Bicinchoninic acid（ BCA） Protein quantitation was done by enzyme linked immunosorbent assay,and i TRAQ（ isobaric tags for relative and absolute quantification） was combined with the 2D-LC-MS / MS analysis method for identification of CYP450 proteins. With Coptidis Rhizoma,30 CYP450 isoenzymes were identified,with 7 proteins significantly increased and 8 decreased in response to Coptidis Rhizoma,while the rest 15 had no change. iTRAQ technology combined with 2D-LCMS / MS method could be used to comprehensively study CYP450 enzyme,but it is necessary to further evaluate in vitro levels of Coptidis Rhizoma and avoid any potential clinical drug-drug interaction.

关 键 词：黄连 同位素标记相对和绝对定量技术(iTRAQ) CYP45同工酶 2D-LC-MS/MS 蛋白质组学 

分 类 号：R284[医药卫生—中药学]