Sane A C, Mendenhall T, Bass D A
Section on Pulmonary, Allergy and Critical Care Medicine, The Bowman Gray School of Medicine of Wake Forest University, Winston Salem, North Carolina 27157-1054, USA.
J Leukoc Biol. 1996 Dec;60(6):704-9. doi: 10.1002/jlb.60.6.704.
Arachidonic acid (AA), the precursor of eicosanoids, is released from the sn-2 position of phospholipids by both secretory (sPLA2) and cytosolic phospholipase A2 (cPLA2). Eicosanoids have been shown to contribute to bronchospasm in asthma. We measured the enzymatic activity of sPLA2 and cPLA2 in the bronchoalveolar lavage fluid and cells, respectively, in male Hartley guinea pigs sensitized with ovalbumin. sPLA2 activity was also measured from alveolar macrophages (AM) in culture from unsensitized and sensitized animals. There was an increase in sPLA2 activity and AA content in the lavage fluid following sensitization (18.73 +/- 1.33 to 25.74 +/- 3.22% hydrolysis and 17.97 +/- 12.39 to 44.76 +/- 13.37 pmol AA/mL BAL, mean +/- SD), which remained elevated but without further increase 4 or 24 h after antigen challenge. AM from unsensitized and sensitized-unchallenged animals did not secrete sPLA2 activity in culture for 3 h and therefore do not appear to be the cell source of the sPLA2 activity present in the alveolar lavage fluid following OA sensitization. In contrast to the increase in sPLA2 in lung lavage fluid, Western blotting for cPLA2 from lung lavage cells showed no increase 4 or 24 h after antigen challenge compared with sensitization alone. cPLA2 enzymatic activity of the cytosol fraction of lung lavage cells showed no changes with antigen sensitization or challenge. In summary, intraperitoneal sensitization with ovalbumin in male Hartley guinea pigs caused an increase in both sPLA2 and AA in bronchoalveolar lavage fluid without a need for antigen challenge. The increased sPLA2 enzymatic activity following sensitization may be responsible for the elevation of AA in the bronchoalveolar lavage fluid observed after antigen sensitization.
花生四烯酸(AA)是类花生酸的前体,可通过分泌型磷脂酶A2(sPLA2)和胞质磷脂酶A2(cPLA2)从磷脂的sn-2位释放出来。类花生酸已被证明与哮喘中的支气管痉挛有关。我们分别测量了用卵清蛋白致敏的雄性Hartley豚鼠支气管肺泡灌洗液和细胞中sPLA2和cPLA2的酶活性。还从未致敏和致敏动物培养的肺泡巨噬细胞(AM)中测量了sPLA2活性。致敏后灌洗液中sPLA2活性和AA含量增加(水解率从18.73±1.33%增至25.74±3.22%,AA含量从17.97±12.39 pmol AA/mL BAL增至44.76±13.37 pmol AA/mL BAL,均值±标准差),抗原激发后4或24小时该值仍保持升高但未进一步增加。未致敏和未激发的致敏动物的AM在培养3小时内未分泌sPLA2活性,因此似乎不是卵清蛋白致敏后肺泡灌洗液中sPLA2活性的细胞来源。与肺灌洗液中sPLA2增加相反,肺灌洗细胞cPLA2的蛋白质印迹显示,与仅致敏相比,抗原激发后4或24小时无增加。肺灌洗细胞胞质部分的cPLA2酶活性在抗原致敏或激发后无变化。总之,雄性Hartley豚鼠腹腔注射卵清蛋白致敏导致支气管肺泡灌洗液中sPLA2和AA均增加,无需抗原激发。致敏后sPLA2酶活性增加可能是抗原致敏后支气管肺泡灌洗液中AA升高的原因。