摘要
目的建立反相高效液相色谱(RP-HPLC)法测定4-羟基-3-甲氧基苯甲酸含量和观察间苯三酚对儿茶酚氧位甲基转移酶(COMT)的抑制作用。方法利用COMT催化3,4-二羟基苯甲酸生成4-羟基-3-甲氧基苯甲酸的原理,通过提取小鼠肝脏匀浆中的COMT 20μL,在反应体系中分别加入1×10-3mol·L-1邻苯二酚及5×10-3,1×10-3,2×10-4mol·L-1间苯三酚各10μL,利用RP-HPLC法测定产物生成量来分析间苯三酚对COMT的抑制作用。结果间苯三酚在5×10-3,1×10-3,2×10-4mol·L-13个浓度下对COMT活性均有抑制作用,抑制率分别为25.3%,17.6%,8.9%。结论间苯三酚对COMT活性具有抑制作用,但抑制作用弱于同浓度的邻苯二酚。
Objective To detect content of 4-hydroxy-3-methoxy benzoic acid by RP-HPLC and observe the inhibitory effect of phloroglucinol on catecholamine-O-methyl transferase（ COMT）. Methods This study used the principle of 3,4-dihydroxy benzoic acid transforming to 4-hydroxy-3-methoxy benzoic acid under COMT＇s catalytic action. COMT（20 μL） was extracted from mouse liver homogenate. In a reaction system,10 μL catecol（1 × 10^-3mol·L^-1 and 10 μL phloroglucinol（5 ×10^-3,1 ×10^-3and 2 × 10^-4mol·L^-1respectively） were added. Products were determined by RP-HPLC to analyze effects of 4-hydroxy-3-methoxy benzoic acid on COMT. Results Phloroglucinol had inhibitory effect on COMT activity at concentrations of 5× 10^-3mol·L^-11 × 10^-3mol·L^-1nd 2 × 10^-4mol · L-1,with the inhibition rate being 25. 3%,17. 6% and 8. 9%,respectively. Conclusion Phloroglucinol has an inhibitory effect on COMT activity,which is weaker than the effect of catechol of the same concentration.
出处
《医药导报》
CAS
北大核心
2014年第10期1303-1306,共4页
Herald of Medicine
