Cigarette smoke extract induces endothelin-1 via protein kinase C in pulmonary artery endothelial cells.
We examined the mechanism of endothelin (ET)-1 regulation by cigarette smoke extract (CSE) and the effect of platelets on CSE-induced stimulation of ET-1 gene expression in human and bovine pulmonary artery endothelial cells (PAECs). Our data show that CSE (1%) induces ET-1 gene expression (after 1 h) and ET-1 peptide synthesis (after 4 h) in bovine PAECs. The induction of preproET-1 mRNA level was due to de novo transcription, and new protein synthesis was not required for this induction. The protein kinase C inhibitors staurosporine (10(-8) mol/l) and calphostin C (10(-7) mol/l) abolished the induction of ET-1 gene expression by CSE in bovine and human PAECs. Although a lower concentration of platelets (10(6) cells/ml in bovine PAECs; 10(7) cells/ml in human PAECs) did not significantly alter ET-1 gene expression in PAECs, incubation of platelets with CSE (1%) and PAECs produced a significant increase in preproET-1 mRNA and ET-1 peptide compared with the values in the presence of CSE (1%) alone. CSE (1%) induced platelet aggregation and increased the expression of platelet membrane glycoproteins ex vivo. Thus our data suggest that CSE stimulates ET-1 gene expression via PKC in PAECs. CSE and platelets showed a synergistic effect on ET-1 gene expression, possibly through the activation of platelets by CSE.
Lee
S D
SD
Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, Asan Medical Center, College of Medicine Ulsan University, Songpa-gu, 138-736 Seoul, Korea. sdlee@www.amc.seoul.kr
Lee
D S
DS
Chun
Y G
YG
Shim
T S
TS
Lim
C M
CM
Koh
Y
Y
Kim
W S
WS
Kim
D S
DS
Kim
W D
WD
D000818:Animals; D001792:Blood Platelets / Q000502:physiology; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D019332:Endothelin-1 / Q000235:genetics / Q000378:metabolism*; D016232:Endothelins / Q000235:genetics; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D015870:Gene Expression / Q000502:physiology; D006801:Humans; D008875:Middle Aged; D009538:Nicotine / Q000494:pharmacology; D010947:Plants, Toxic; D010974:Platelet Aggregation; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism*; D011498:Protein Precursors / Q000235:genetics; D011651:Pulmonary Artery / Q000166:cytology / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D012906:Smoke; D014026:Nicotiana
D019332:Endothelin-1; D016232:Endothelins; D010980:Platelet Membrane Glycoproteins; D011498:Protein Precursors; D012333:RNA, Messenger; D012906:Smoke; D009538:Nicotine; D011493:Protein Kinase C