HPLC法测定脑梗死患者血清PDE活性及临床意义  

Clinical significance of determination of activity of serum PDE in cerebral infarction patients by HPLC.

作　　者：林月好[1] 吴婉玲[2] 姚晓黎[2] 李才明[3] 莫照龙[4] 苏全喜[5] 张为西[2] 卢锡林[2] 

机构地区：[1]中山大学附属肿瘤医院检验科,广东广州510060 [2]中山大学附属第一医院神经科,广东广州510080 [3]惠州市中心人民医院,广东惠州516001 [4]海南省农垦总局医院,海南海口570311 [5]广东药学院第一附属医院神经科,广东广州510080

出　　处：《中国热带医学》2008年第11期1949-1950,2022,共3页China Tropical Medicine

基　　金：广东省博士启动基金资助项目(编号:5300783;04300353);广东省科技计划项目(编号:2006B36003003);广东省医学科研基金(编号:B2001121);海南省自然科学基金(编号:30325)

摘　　要：目的建立血清磷酸二酯酶(PDE)活性测定的HPLC方法并探讨其在脑梗死患者的临床意义。方法分离293例脑梗死患者和148例对照组外周血白细胞,匀浆可溶部分为酶样品,以cAMP为反应底物,采用底物的减少来测定PDE的活性。色谱柱为Nova-Pak18C(3.9 mm×150 mm,4μm),流动相为甲醇-磷酸盐缓冲液(10:90,V/V);流速为0.8mL/min,柱温为室温;检测波长为254nm。结果PDE底物(cAMP、cGMP)和系列产物(5’AMP、5’GMP、腺苷、次黄嘌呤和腺嘌呤)分离良好,无干扰。cAMP和cGMP检测限分别为0.125μmoL/L和0.0625μmoL/L;平均回收率91.2%～102.8%;日内及日间RSD均<10%。脑梗死组外周血PDE活性为(13.805±3.121),对照组为(7.334±2.324),差异有统计学意义(P<0.001)。结论脑梗死患者PDE活性增高;HPLC法简单易行,能精确可靠地测定外周血白细胞的PDE活性。Objective To establish a high performance liquid chromatography （HPLC） method for determining phosphodiesterase activity in serum of ischemic stroke（IS） patients and approach the clinical significance. Methods There 293 IS patients and 148 controls were recruited in this study, their PDE activities were detected using HPLC. The WBC were separate and the parts of dissolution were serviced as enzyme samples, cyclic adenosine monophosphate （cAMP） and cyclic guanylic acid（cGMP） were used as substrates, using double -tubes method to determine phosphodiesterase （PDE） activities. Nova- Pak 18C （3.9mm × 150mm,4 μm） column was used. The mobile phase consisted of methyl alcohol and phosphate buffer （ 10：90, V/V）. The flow rate of mobile phase was 0.8 mL/min ; the UV detection wave length was 254 nm. Results The contents of cAMP, cGMP, 5＇ AMP, 5＇ GMP, adenosine, hypoxanthine and adenine were separated well and not interfered. The lowest detectable limit of cAMP and cGMP was 0. 125 μmoL/L and 0. 0625 μmoL/L respectively. The average recovery was 91.2% - 102.8%. The RSD of within - day and between - day was less than 10%. PDE activities in IS patients （ 13. 805 ±3.121 ） were higher than those in controls （7. 334 ±2. 324） （P 〈 0. 001 ）. Conclusion The activity of PDE in IS patients is higher than those in controls. HPLC is a convenient, sensitive and accurate method for determining PDE activity.

关 键 词：脑梗死 磷酸二酯酶 高效液相色谱法 

分 类 号：R743[医药卫生—神经病学与精神病学]