摘要
目的 建立一种用同一底物检测分泌型磷脂酶A2 (sPLA2 )和胞浆型磷脂酶A2 (cPLA2 )的放射化学方法。方法 用3 H 花生四烯酸参入大肠杆菌膜 ,在一定条件下被待测PLA2 水解 ,以水解率的大小表示酶活性的高低。结果 测定cPLA2 和sPLA2 反应体系中最适pH值分别是 7.8和 8 0 ,最适钙离子浓度是 5和 13mmol/L ;cPLA2 批内CV 5 .2 % ,批间CV 10 .9% ,sPLA2 则分别是 4.9%和 7.8% ;倍比稀释实验结果具有良好的线性关系 ;急性胆囊炎患者血清sPLA2 活性明显高于正常人 ,内毒素 (LPS)诱导前后白血病K5 62细胞内和培养液中PLA2 活性差异有显著性 (P <0 .0 5 )。
Objective To develop a new radiochemistry method to assay the secretory phospholipase A 2(sPLA 2) and cytosolic phospholipase A 2 (cPLA 2) with a same substrate. Methods E.coli membrane doped by 3H arachidonic acid was prepared and hydrolyzed by PLA 2 in certain condition, and the enzyme activity was expressed with the hydrolyzing rate. Results Intra day coefficient of variation( CV ) of cPLA 2 was 5.2% and inter day CV was 10.9%, and 4.9% and 7.8% for sPLA 2 respectively. Results of a series proportional dilution assay showed a good linear relationship. Serum sPLA 2 activities of patients with acute cholecystitis were significantly higher than that of normal control subjects. There was a significant difference of activities of sPLA 2 and cPLA 2 between the endotoxin induced leukemia cell K562 and control. Conclusions This method is specific, stable and sensitive, it may be used in clinical and scientific research.
出处
《中华核医学杂志》
CAS
CSCD
北大核心
2001年第4期242-243,共2页
Chinese Journal of Nuclear Medicine
基金
重庆市教委重点科研项目基金资助 (995 2 )
