Urothelial cell platelet-activating factor production mediated by calcium-independent phospholipase A2γ.
Urothelial cell platelet-activating factor production mediated by calcium-independent phospholipase A2γ.
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DOI：
10.1016/j.urology.2010.08.032
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发表时间：
2011-01
期刊：
影响因子：
2.1
通讯作者：
McHowat J
中科院分区：
文献类型：
--
作者：
Rastogi P;Rickard A;Klumpp DJ;McHowat J
确定磷脂酶A2（PLA 2）抑制剂对类胰蛋白酶刺激下尿路上皮细胞血小板活化因子（PAF）产生的影响。从正常人输尿管分离的尿路上皮细胞用HPV 16 E6 E7型永生化（TEU-2细胞）。在存在和不存在钙的情况下，使用（16：0，[3 H]18：1）血浆胆碱和磷脂酰胆碱底物测量TEU-2细胞中的PLA 2活性。在用[3 H]乙酸预标记的TEU-2细胞中测量了[3 H]PAF的产生。通过测定与TEU-2细胞蛋白孵育的[3 H]PAF水解的[3 H]乙酸的量来测量PAF-乙酰水解酶活性。通过测量与TEU-2细胞孵育后粘附的PMN中的髓过氧化物酶活性来评估人PMN对TEU-2细胞的粘附。在TEU-2细胞中测量的大多数PLA 2活性被确定为膜相关的、钙非依赖性的（iPLA 2）并且对胞浆胆碱底物具有选择性。用类胰蛋白酶刺激TEU-2细胞导致血小板活化因子（PAF）的产生增加和多形核白细胞（PMN）粘附增加，而用iPLA 2 γ-选择性抑制剂（R）-溴烯醇内酯（（R）-BEL）预处理可完全抑制。用胞质PLA 2抑制剂甲基花生四烯酰氟膦酸酯（MAFP）预处理导致类胰蛋白酶刺激的PAF产生和PMN粘附TEU-2细胞的增强，这是PAF乙酰水解酶（PAF-AH）抑制的结果。TEU-2细胞的类胰蛋白酶刺激导致iPLA 2 γ的活化，导致PAF产生增加和PMN粘附增加。用MAFP抑制TEU-2细胞PAF-AH活性增强类胰蛋白酶刺激的PAF产生和PMN粘附。
To determine the effect of phospholipase A2 (PLA2) inhibitors on urothelial cell platelet-activating factor (PAF) production in response to tryptase stimulation. Urothelial cells isolated from normal human ureters were immortalized with HPV type 16E6E7 (TEU-2 cells). PLA2 activity in TEU-2 cells was measured using (16:0, [3H]18:1) plasmenylcholine and phosphatidylcholine substrates in the presence and absence of calcium. [3H]PAF production was measured in TEU-2 cells prelabeled with [3H] acetic acid. PAF-acetylhydrolase activity was measured by determine the amount of [3H] acetate hydrolysed from [3H]PAF incubated with TEU-2 cellular protein. Adherence of human PMN to TEU-2 cells was assessed by measuring myeloperoxidase activity in adherent PMN following incubation with TEU-2 cells. The majority of PLA2 activity measured in TEU-2 cells was determined to be membrane-associated, calcium-independent (iPLA2) and selective for plasmenylcholine substrate. Stimulation of TEU-2 cells with tryptase results in increased production of platelet-activating factor (PAF) and increased polymorphonuclear leukocyte (PMN) adherence that were inhibited completely by pretreatment with the iPLA2γ-selective inhibitor (R)-bromoenol lactone ((R)-BEL). Pretreatment with the cytosolic PLA2 inhibitor methyl arachidonyl fluorophosphonate (MAFP) results in potentiation of tryptase-stimulated PAF production and PMN adherence to TEU-2 cells that is a result of PAF-acetylhydrolase (PAF-AH) inhibition. Tryptase stimulation of TEU-2 cells results in activation of iPLA2γ leading to an increase in PAF production and increased PMN adherence. Inhibition of TEU-2 cell PAF-AH activity with MAFP potentiated tryptase-stimulated PAF production and PMN adherence.
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影响因子：
--
作者：
Joannidis, M;Truebsbach, S;Wiedermann, CJ
通讯作者：
Wiedermann, CJ
影响因子：
2.5
作者：
Iatrou, C;Moustakas, G;Ziroyiannis, P
通讯作者：
Ziroyiannis, P
DOI：
10.1152/ajprenal.00072.2002
发表时间：
2002-11-01
影响因子：
4.2
作者：
Rickard, A;McHowat, J
通讯作者：
McHowat, J
DOI：
10.1161/01.atv.0000016153.47693.b2
发表时间：
2002-05-01
影响因子：
8.7
作者：
Prescott, SM;McIntyre, TM;Stafforini, DM
通讯作者：
Stafforini, DM
影响因子：
--
作者：
UGAILYTHULESIUS, L;THULESIUS, O;SABHA, M
通讯作者：
SABHA, M