Nevalainen T J
Department of Pathology, University of Turku, Finland.
Clin Chem. 1993 Dec;39(12):2453-9.
Phospholipase A2 (EC 3.1.1.4; PLA2) is detected in serum by determination of either the catalytic activity of the enzyme or the concentration of the enzyme protein by immunoassays. The most sensitive methods for determining PLA2 catalytic activity are radiometric assays, with a substrate of synthetic phospholipid (e.g., phosphatidylcholine or phosphatidylethanolamine) containing a 14C- or 3H-labeled fatty acid at the sn-2-position. Membranes of autoclaved Escherichia coli grown in the presence of radioactive oleic acid may also be used as a substrate. The released fatty acids are separated from the unreacted substrate and quantified by liquid scintillation counting. PLA2 catalytic activities are increased in serum in sepsis, acute pancreatitis, peritonitis, multiple injuries, rheumatoid arthritis, and other arthropathies. Immunoassays--radioimmunoassay, enzyme-linked immunosorbent assay, or time-resolved fluoroimmunoassay--are based on the use of either polyclonal or monoclonal antibodies to purified PLA2s. Specific assays have been developed for both pancreatic group I PLA2 (PLA2-I) and nonpancreatic group II PLA2 (PLA2-II). The cellular source of PLA2-I in serum is the pancreatic acinar cell. Increased serum PLA2-I values have been reported in acute pancreatitis, pancreatic cancer, and abdominal trauma. Increased PLA2-II values are found in conditions involving inflammation, e.g., sepsis, infections, acute pancreatitis, various forms of arthritis, cancer, complications of pregnancy, and postoperative states. Good correlations have been found in serum samples between the catalytic activity of PLA2 and the concentration of PLA2-II but not PLA2-I. PLA2-II may represent an acute-phase protein. The cellular source of the PLA2-II in serum is unknown; it is present in large amounts in cartilage and Paneth cells, prostatic gland cells, seminal fluid, lacrimal gland cells, and tears, but cannot be demonstrated by immunohistochemical or immunochemical methods in inflammatory cells.
通过测定酶的催化活性或采用免疫测定法测定酶蛋白浓度,可在血清中检测到磷脂酶A2(EC 3.1.1.4;PLA2)。测定PLA2催化活性最灵敏的方法是放射性测定法,其底物为在sn-2位含有14C或3H标记脂肪酸的合成磷脂(如磷脂酰胆碱或磷脂酰乙醇胺)。在放射性油酸存在下生长的高压灭菌大肠杆菌的膜也可用作底物。释放出的脂肪酸与未反应的底物分离,并通过液体闪烁计数进行定量。在败血症、急性胰腺炎、腹膜炎、多发伤、类风湿关节炎和其他关节病中,血清中的PLA2催化活性会升高。免疫测定法——放射免疫测定法、酶联免疫吸附测定法或时间分辨荧光免疫测定法——是基于使用针对纯化的PLA2的多克隆或单克隆抗体。已针对胰腺I型PLA2(PLA2-I)和非胰腺II型PLA2(PLA2-II)开发了特异性测定法。血清中PLA2-I的细胞来源是胰腺腺泡细胞。据报道,急性胰腺炎、胰腺癌和腹部创伤时血清PLA2-I值会升高。在涉及炎症的情况下,如败血症、感染、急性胰腺炎、各种形式的关节炎、癌症、妊娠并发症和术后状态,可发现PLA2-II值升高。在血清样本中,已发现PLA2的催化活性与PLA2-II的浓度之间具有良好的相关性,但与PLA2-I无相关性。PLA2-II可能代表一种急性期蛋白。血清中PLA2-II的细胞来源尚不清楚;它大量存在于软骨和潘氏细胞、前列腺细胞、精液、泪腺细胞和泪液中,但在炎症细胞中无法通过免疫组织化学或免疫化学方法证实其存在。