Effect of silica on phospholipase D activity in rat alveolar macrophages.
Silica may act as a stimulator of pulmonary inflammation and fibrosis. The effect of silica on phospholipase D (PLD) activity assayed as accumulation of [3H]phosphatidylethanol ([3H]PtdEt) was examined in [3H]palmitic acid-labeled primary cultures of rat alveolar macrophages. Silica induced a rapid accumulation of [3H]PtdEt in a time (0, 15, 30 and 45 min)- and concentration (0.5, 1.0, 2.5 and 5.0 mg/ml)-dependent manner indicating PLD activation. This silica-stimulated PLD activity was attenuated by the pretreatment with calcium chelator ethylene glycol-bis(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA) or/and 1,2-bis(2-aminophenoxy)ethane-N,N,N,N-tetraacetic acid acetoxymethyl ester (BAPTA/AM) (EGTA: 54.3 +/- 8.6%, BAPTA/AM: 67.5 +/- 7.8% and EGTA + BAPTA/AM: 35.8 +/- 2.9, respectively). Also, silica-induced PLD activation was partially inhibited by the pretreatment with nonspecific phospholipase C (PLC) and PLD inhibitor (neomycin; 66.4 +/- 4.8%) or specific PLC inhibitor (U73122; 70.8 +/- 4.6%). Sphingosine as a protein kinase C (PKC) inhibitor did not change silica-induced PLD activity indicating that PKC might not play a role in PLD activation by silica. Based on these results, we concluded that a silica-stimulated phospholipase D activity is present in the rat alveolar macrophages and is predominantly regulated by PLC-mediated intracellular calcium.
Cha
S H
SH
Department of Pharmacology, Catholic University Medical College, Seoul, Korea.
Lee
W K
WK
Kim
K A
KA
Lim
Y
Y
Han
J S
JS
Lee
K H
KH
D000704:Analysis of Variance; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D016676:Macrophages, Alveolar / Q000187:drug effects* / Q000201:enzymology*; D008297:Male; D010739:Phospholipase D / Q000187:drug effects* / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012822:Silicon Dioxide / Q000494:pharmacology*; D013110:Sphingosine / Q000494:pharmacology; D010738:Type C Phospholipases / Q000378:metabolism
D002614:Chelating Agents; D004533:Egtazic Acid; D012822:Silicon Dioxide; D010738:Type C Phospholipases; D010739:Phospholipase D; C025603:1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; D013110:Sphingosine; D002118:Calcium