Pruzanski W, Vadas P, Kim J, Jacobs H, Stefanski E
Immunology Diagnostic and Research Centre, University of Toronto, Wellesley Hospital, Canada.
J Rheumatol. 1988;15(5):791-4.
High activity of phospholipase A2 (PLA2) has been detected in synovial fluids (SF) in inflammatory arthritides. Since the source(s) of SF PLA2 has not been identified, we tested PLA2 content in SF cells obtained from 11 SF. Cell sonicates were prepared at 5 X 10(6) cells/ml. In the supernatants of the sonicated SF cells (n = 11), PLA2 activity ranged from 38-755 U/ml, mean 368 +/- 243 (SD) U/ml, compared to 5-64 U/ml, mean 31 +/- 15 (SD) U/ml in sonicates of normal peripheral blood PMN (n = 5) (p less than 0.0001). Spontaneous release of PLA2 from unstimulated SF cells ranged from 26-365 U/ml, mean 131 +/- 144 (SD) U/ml (n = 5), whereas spontaneous release from peripheral blood PMN was negligible. Neither 10(-8) M FMLP nor 5 X 10(-6) M dexamethasone altered extracellular PLA2 release. To assess whether PLA2 adsorbs passively to cell membranes through hydrophobic interaction, normal peripheral PMN were incubated in crude SF (n = 7) with PLA2 ranging from 4,000-24,300 U/ml, or with purified human SF PLA2 or Naja naja PLA2. We found that soluble PLA2 adsorbed to PMN membranes in a concentration dependent fashion. PLA2 activity in sonicates of PMN incubated in crude SF ranged from 185-358 U/ml compared to controls of 21-64 U/ml. Sonicates of PMN incubated with purified human SF PLA2 (5,000-30,000 U/ml) showed progressive concentration dependent increase in PLA2 from 7 +/- 4 to 212 +/- 11 (SD) U/ml (p less than 0.001). PMN incubated with Naja naja PLA2 also showed marked increase in the content of PLA2.(ABSTRACT TRUNCATED AT 250 WORDS)
在炎性关节炎的滑液(SF)中已检测到磷脂酶A2(PLA2)的高活性。由于尚未确定SF PLA2的来源,我们检测了从11份SF中获取的SF细胞中的PLA2含量。细胞超声匀浆以5×10⁶个细胞/毫升的浓度制备。在超声处理的SF细胞的上清液中(n = 11),PLA2活性范围为38 - 755 U/毫升,平均为368±243(标准差)U/毫升,而正常外周血中性粒细胞(PMN)的超声匀浆(n = 5)中为5 - 64 U/毫升,平均为31±15(标准差)U/毫升(p < 0.0001)。未刺激的SF细胞中PLA2的自发释放范围为26 - 365 U/毫升,平均为131±144(标准差)U/毫升(n = 5),而外周血PMN的自发释放可忽略不计。10⁻⁸ M的N-甲酰甲硫氨酰-亮氨酰-苯丙氨酸(FMLP)和5×10⁻⁶ M的地塞米松均未改变细胞外PLA2的释放。为评估PLA2是否通过疏水相互作用被动吸附到细胞膜上,将正常外周PMN在粗制SF(n = 7)中与PLA2浓度范围为4000 - 24300 U/毫升孵育,或与纯化的人SF PLA2或眼镜蛇毒PLA2孵育。我们发现可溶性PLA2以浓度依赖性方式吸附到PMN膜上。在粗制SF中孵育的PMN超声匀浆中的PLA2活性范围为185 - 358 U/毫升,而对照组为21 - 64 U/毫升。与纯化的人SF PLA2(5000 - 30000 U/毫升)孵育的PMN超声匀浆显示PLA2从7±4逐渐增加到212±11(标准差)U/毫升(p < 0.001)。与眼镜蛇毒PLA2孵育的PMN也显示PLA2含量显著增加。(摘要截断于250字)