Nguyen H T, Rice G E, Farrugia W, Wong M, Brennecke S P
Department of Physiology, Monash University, Clayton, Victoria, Australia.
Biol Reprod. 1994 Mar;50(3):526-34. doi: 10.1095/biolreprod50.3.526.
The aim of this study was to establish the effect of bacterial endotoxin lipopolysaccharide (LPS) on type II phospholipase A2 (PLA2) content and in vitro net PLA2 enzymatic activity in human choriodecidua. More particularly, the objective was to ascertain whether an increase in type II PLA2 tissue content and PLA2 enzymatic activity is associated with the previously documented stimulatory effect of LPS on prostaglandin E2 (PGE2) release from choriodecidua. Choriodecidua explants were incubated in RPMI 1640 (control) or RPMI 1640 containing LPS (0.1 ng/ml to 10 micrograms/ml) for up to 24 h. Under the incubation conditions utilized, LPS (1 microgram/ml) stimulated the release of PGE2 3-fold (p < 0.001, n = 4 tissues), in a time-dependent manner (p < 0.005). Tissue immunoreactive (ir)-type II PLA2 content and PLA2 enzymatic activity were determined by monoclonal antibody sandwich ELISA and radiolabeled enzyme assay procedures, respectively. The tissue content of type II PLA2 immunoreactivity in choriodecidua obtained from women (n = 7) at term but not in labor averaged 155 +/- 24 ng/ml DNA. PLA2 enzymatic activity average 29.8 +/- 5.1 pmol phosphatidylethanolamine (PE)/mg DNA/h. LPS at a concentration of 1 microgram/ml increased immunoreactive type II PLA2 tissue content by 180% of the control value (p < 0.05, n = 7) within 1 h of exposure and remained significantly elevated for 18 h when compared to controls (174% of control, p < 0.01, n = 7).(ABSTRACT TRUNCATED AT 250 WORDS)
本研究的目的是确定细菌内毒素脂多糖(LPS)对人绒毛膜蜕膜中II型磷脂酶A2(PLA2)含量及体外净PLA2酶活性的影响。更具体地说,目标是确定II型PLA2组织含量和PLA2酶活性的增加是否与先前记录的LPS对绒毛膜蜕膜中前列腺素E2(PGE2)释放的刺激作用相关。将绒毛膜蜕膜外植体在RPMI 1640(对照)或含LPS(0.1 ng/ml至10 μg/ml)的RPMI 1640中孵育长达24小时。在所采用的孵育条件下,LPS(1 μg/ml)以时间依赖性方式刺激PGE2释放3倍(p < 0.001,n = 4个组织)(p < 0.005)。分别通过单克隆抗体夹心ELISA和放射性标记酶测定程序测定组织免疫反应性(ir)-II型PLA2含量和PLA2酶活性。从足月但未临产的妇女(n = 7)获得的绒毛膜蜕膜中II型PLA2免疫反应性的组织含量平均为155±24 ng/ml DNA。PLA2酶活性平均为29.8±5.1 pmol磷脂酰乙醇胺(PE)/mg DNA/小时。浓度为1 μg/ml的LPS在暴露1小时内使免疫反应性II型PLA2组织含量比对照值增加180%(p < 0.05,n = 7),与对照相比,在18小时内仍显著升高(为对照的174%,p < 0.01,n = 7)。(摘要截短于250字)