芳香酯酶测定底物探讨及临床应用评价  

Study on assay arylesterase' s substrate and evaluation of clinical applicatio

作　　者：胡望平[1] 周长邵[1] 胡盈莹[2] 黄兢姚[3] 王海林[1] 孙艳[1] 

机构地区：[1]福州军区福州总医院(福建医大福总临床医学院)生化科,福建福州350025 [2]福州市传染病医院肝病研究所,福建福州350025 [3]福建医大协和医院介入科,福建福州350001

出　　处：《中国现代医学杂志》2004年第21期66-68,74,共4页China Journal of Modern Medicine

摘　　要：目的该科合成芳香酯酶(Arylester hydrolase EC 3.1.1.2,ArE)底物4-已酰氧基-3-甲氧基-ω-硝基乙烯;建立手工的和自动生化仪检测方法;评价在常见肝病中的应用.方法底物合成:香草醛加冰乙酸、乙酸铵.在乙醇溶液中用硝基四烷进行硝基化,滤取结晶.用乙酸酐进行酯化,滤取结晶,重结晶.方法建立.程序:取pH8.6Tris-HCL缓冲液2.5 ml,加血清30 ul,再加3mmol/l底物液0.4 ml,37℃10 min,加5 ml/dl正丁醇3 ml.混匀,在520nm,读取吸光度.摩尔消光系数(20345±280)A.计算:ArE U/L=971.3×A.结果反应条件:底物溶剂以甲醇13.65%为最佳.产物光吸收峰498nm,选用520 nm,吸光度达93.20%.择取pH8.6缓冲液,加0.51 mmol/L Ca2+和68.3 mmol/L Na+可提酶活力14.00%.批间CV为3.21%～4.52%.溶血、黄疸和脂血标本对ArE结果无干扰.临床应用:90例健康人ArE(470.6±49.7)U/L,参考值:＜421U/L.肝硬化、肝癌、肝炎3组ArE结果x±S,U/L分别是(266±77),(35±84)和(366±111);ROC曲线下面积分别是0.979,0.910和0.807.面积愈大诊断价值愈高,ArE愈低,肝功能愈差.尤其对肝硬化、肝癌.肝硬化组＜421 U/L阳性率为92.30%;肝癌ArE(Y)与脂碱脂酶CHE相关系数为0.909,Y=0.045X+190.53.结论ArE简便可靠.在肝硬化、肝癌、肝炎中阳性率性高,特异性好.有望成为重要常规肝功指标.Objective:To synthesize a substrate of arylesterase:4-acetoxy-3-methoxy-ω-nitrostyrene in or-der to study on action of arylesterase.To establish a new method for measurement of serum arylesterase ac-tivity and to investigate and to evaluate its clinical application on hepatogenous disease.Methods:To synthe-size a substrate.The first was nitricatio.Ammonium acetate,glacial acetic acid,nitromethane were added to a solution of vanillin,4-hydroxy-3-methoxy-ω-nitrostyrene has been crystallized out.The second was esterize.4-hydroxy-3-methoxy-ω-nitrostyrene was heated with acetic anhydrde for esterize,a yellow product waspre-cipitated and filtered off,it was4-acetoxy-3-methoxy-ω-nitrostyrene.Establish of A Novel Assay method.The procedure for arylesterase:Pipet2.5ml of pH8.6Tris-HCl buffer to a tube.Added30μl serum,add a-gain0.4ml the substrate solution,after the test has reacted for10minutes at37℃,added3ml butanol5ml/dl,mixed thoroughly.Reading absorbance at520nm.Calculation:ArE U/L=971.3×A.Mol extinction coefficient was(20345±280)A.Results:Reaction condition:The methyl alcohol13.65ml/dl has been selected as sub-strate solvent.According to Olympus AU2700analysis appearance,520nm been selected.Buffer pH8.6has been selected after compare the spontaneous hydrolysis and sensitivity.Adding0.51mmol/L Ca 2+ and68.3mmol/L Na+ may be to bring up ArE enzyme's activity 14.00%. The method CV is 3.21%~4.52% in 20 day. It was not interference that hemolisis, jaundice, lipemia serum. Clinical application: Health 90 persons ArE (470.6±49.7) U/L, ArE reference value is <421 U/L. cirrhosis of live, hepatocarcinoma, hepatitis tris series ArE mean x±SD (U/L) was respectively (266±77), (35±84), (366±111); Area under the curve to healthy subjects was separately 0.979, 0.910, 0.807. The area under the curve was more maximus the diagnosis value ever morfine, especially for cirrhosis of liver and hapatocarcinoma. The correlation coefficient was respectively 0.872, 0.840, P <0.01, between ArE of hepatopathy and albumin, CHE.

关 键 词：芳香酯酶底物 肝硬化 肝病 血清芳香酯酶 

分 类 号：R446[医药卫生—诊断学] R575[医药卫生—临床医学]