复方贞术调脂方调节HMG-CoA还原酶活性成分的快速筛选  

A Quick Method for Screening Active Components of Fufang Zhenqi Tiaozhi Prescription on Regulating HMG-CoA Reductase Activity

作　　者：胡旭光[1] 郭姣[1] 贝伟剑[1] 胡因铭[1] 黄利华[1] 曹扬[1] 

机构地区：[1]广东药学院/国家中医药管理局脂代谢三级实验室及高脂血症调肝降脂重点研究室,广州510006

出　　处：《中药新药与临床药理》2010年第5期468-471,共4页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(30873414;30973913);国家科技重大专项(2009zx09103-407;2009B090300349)

摘　　要：目的优化快速筛选影响羟甲基戊二酸单酰辅酶A还原酶(HMG-CoA还原酶)活性成分的方法,并筛选复方贞术调脂方(FTZ)中抑制HMG-CoA还原酶的活性成分。方法利用纯化的猪肝HMG-CoA还原酶,从HMG-CoA还原酶浓度、HMG-CoA和NADPH底物浓度三方面优化HMG-CoA还原酶筛选方法,评价FTZ和FTZ中不同药物成分对HMG-CoA还原酶活性的影响。结果猪肝HMG-CoA还原酶粗酶制剂具有明显的酶促动力学特性,反应体系中30μgHMG-CoA还原酶粗酶、200mmol/LNADPH、6μmol/LHMG-CoA是检测HMG-CoA还原酶活性和筛选影响酶活性成分的优化条件;FTZ和人参皂苷Rb2、三七总皂苷和阳性对照普伐他汀对HMG-CoA还原酶活性有显著的抑制作用,最大抑制率分别为23.7%、44.6%、35.5%、45.7%(P<0.05或P<0.01)。结论 HMG-CoA还原酶是FTZ调节脂代谢的靶点之一,人参皂苷Rb2、三七总皂苷是FTZ中抑制HMG-CoA还原酶的活性成分。Objective To study the new approach for rapid screening the active ingredients which affect the hydroxymethyl glutaryl coenzyme A（HMG-CoA）reductase activity from Chinese herbal medicine of Fufang Zhenqi Tiaozhi（FZT）Prescription.Methods Purified HMG-CoA reductase isolated from normal pig liver homogenate and colorimetric assays were used to investigate the effects of FZT and different components of FZT on HMG-CoA reductase activity with the concentrations of HMG-CoA,HMG-CoA reductase and NADPH substrate as evaluation parameters.Results Normal pig liver crude HMG-CoA reductase preparation in the pH7.0 buffer solution had the significant enzymatic kinetic properties of HMG-CoA reductase,The reaction system containing 200 mmol/L NADPH,30 μg HMG-CoA reductase,6 μmol/L HMG-CoA was suitable for the evaluation of the inhibition of HMG-CoA reductase activity by Chinese herbal ingredients.FZT,ginsenoside Rb2,saponins of Radix Notoginseng and positive control of pravastatin had obvious inhibition on HMG-CoA reductase activity,the maximum inhibitive rate being 23.7 %,44.6 %,35.5 %,45.7% respectively（P 0.05 or P 0.01）.Conclusion A simple,rapid method for screening the active ingredient of Chinese herbal medicine affected HMG-CoA reductase activity has been established.It is indicated that HMG-CoA reductase is one of the targets of FZT for regulating lipid metabolism,and ginsenoside Rb2 and saponins of Radix Notoginseng are the active ingredients from FZT for inhibiting HMG-CoA reductase activity.

关 键 词：复方贞术调脂方 羟甲基戊二酸单酰辅酶A还原酶 调脂活性成分 

分 类 号：R285.5[医药卫生—中药学]