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...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。展开更多
A lab-made off line 2D-LC-MS system was developed for protein analysis. In case the off line 2D-LC was utilized for peptide separation, and the working conditions for strong cation exchange(SCX) chromatography and the...A lab-made off line 2D-LC-MS system was developed for protein analysis. In case the off line 2D-LC was utilized for peptide separation, and the working conditions for strong cation exchange(SCX) chromatography and the succeeding reversed phase(RP) chromatography can be individually optimized, contrasted to the on-line system. The experimental results show that the mass fractions of acetonitrile for the SCX separation can be increased form 5% as references reported up to 20% so that the separation of peptides has been substantially improved. The off line system has been applied to the analysis of proteins extracted from the cancer tissue of nude mice inoculated by a human-hepatocellular-carcinoma cell-line D20. Some extreme basic proteins(pI>10) and larger proteins(M w>200 000), which are difficult to be isolated by the traditional 2D-PAGE method, have been identified in D20 sample.展开更多
文摘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。
文摘A lab-made off line 2D-LC-MS system was developed for protein analysis. In case the off line 2D-LC was utilized for peptide separation, and the working conditions for strong cation exchange(SCX) chromatography and the succeeding reversed phase(RP) chromatography can be individually optimized, contrasted to the on-line system. The experimental results show that the mass fractions of acetonitrile for the SCX separation can be increased form 5% as references reported up to 20% so that the separation of peptides has been substantially improved. The off line system has been applied to the analysis of proteins extracted from the cancer tissue of nude mice inoculated by a human-hepatocellular-carcinoma cell-line D20. Some extreme basic proteins(pI>10) and larger proteins(M w>200 000), which are difficult to be isolated by the traditional 2D-PAGE method, have been identified in D20 sample.
