Abstract
We examined the effect of nitric oxide (NO) on cell adhesion using cultured human pulmonary microvascular endothelial cells (PMVEC). Attachment of these cells to fibronectin was significantly inhibited by NO donors, spermine NONOate and S-nitroso-N-acetyl-penicillamine or L-arginine, but not 8-bromoguanosine-3',5'-cyclic-monophosphate. Similar results were obtained with the electrical cell-substrate impedance sensor (ECIS) technique. Addition of NO donors or L-arginine, but not 8-bromoguanosine-3',5'-cyclic- monophosphate or N2,2'-O-dibutyrylguanosine-3',5'-cyclic-monophosphate, to confluent PMVEC monolayers resulted in a transient decrease in cell adhesion, which was quantitated by the ECIS. Exposure to 1 U/ml α-thrombin reduced the monolayer electrical resistance by ~50%. The observed response was significantly suppressed by pretreatment of cells with intracellular calcium chelator, 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid or NO synthase inhibitor, N(G)-nitro-L-arginine methyl ester, but not guanylate cyclase inhibitor, 6-anilino-5,8-quinoline-quinone. Selective knockout of endothelial NO synthase with antisense oligodeoxynucleotides also significantly reduced thrombin-induced decrease in monolayer resistance. Our findings indicate that thrombin stimulates calcium-dependent release of NO from PMVEC, which mediates the retraction of endothelial cells via a cGMP- independent pathway. Our results suggest that NO modulates cell-matrix and/or cell-cell adhesion in PMVEC and that this molecule might modify microvascular permeability in the human lung. (C) 2000 Elsevier Science Inc.
| Original language | English |
|---|---|
| Pages (from-to) | 1-11 |
| Number of pages | 11 |
| Journal | Life Sciences |
| Volume | 67 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - May 26 2000 |
| Externally published | Yes |
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Keywords
- Cell adhesion
- Electrical cell-substrate impedance sensor
- Human
- Nitric oxide
- Pulmonary microvascular endothelial cell
ASJC Scopus subject areas
- Pharmacology
Cite this
Role of nitric oxide in human pulmonary microvascular endothelial cell adhesion. / Tsukahara, Hirokazu; Noiri, Eisei; Jiang, Mi Zu; Hiraoka, Masahiro; Mayumi, Mitsufumi.
In: Life Sciences, Vol. 67, No. 1, 26.05.2000, p. 1-11.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Role of nitric oxide in human pulmonary microvascular endothelial cell adhesion
AU - Tsukahara, Hirokazu
AU - Noiri, Eisei
AU - Jiang, Mi Zu
AU - Hiraoka, Masahiro
AU - Mayumi, Mitsufumi
PY - 2000/5/26
Y1 - 2000/5/26
N2 - We examined the effect of nitric oxide (NO) on cell adhesion using cultured human pulmonary microvascular endothelial cells (PMVEC). Attachment of these cells to fibronectin was significantly inhibited by NO donors, spermine NONOate and S-nitroso-N-acetyl-penicillamine or L-arginine, but not 8-bromoguanosine-3',5'-cyclic-monophosphate. Similar results were obtained with the electrical cell-substrate impedance sensor (ECIS) technique. Addition of NO donors or L-arginine, but not 8-bromoguanosine-3',5'-cyclic- monophosphate or N2,2'-O-dibutyrylguanosine-3',5'-cyclic-monophosphate, to confluent PMVEC monolayers resulted in a transient decrease in cell adhesion, which was quantitated by the ECIS. Exposure to 1 U/ml α-thrombin reduced the monolayer electrical resistance by ~50%. The observed response was significantly suppressed by pretreatment of cells with intracellular calcium chelator, 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid or NO synthase inhibitor, N(G)-nitro-L-arginine methyl ester, but not guanylate cyclase inhibitor, 6-anilino-5,8-quinoline-quinone. Selective knockout of endothelial NO synthase with antisense oligodeoxynucleotides also significantly reduced thrombin-induced decrease in monolayer resistance. Our findings indicate that thrombin stimulates calcium-dependent release of NO from PMVEC, which mediates the retraction of endothelial cells via a cGMP- independent pathway. Our results suggest that NO modulates cell-matrix and/or cell-cell adhesion in PMVEC and that this molecule might modify microvascular permeability in the human lung. (C) 2000 Elsevier Science Inc.
AB - We examined the effect of nitric oxide (NO) on cell adhesion using cultured human pulmonary microvascular endothelial cells (PMVEC). Attachment of these cells to fibronectin was significantly inhibited by NO donors, spermine NONOate and S-nitroso-N-acetyl-penicillamine or L-arginine, but not 8-bromoguanosine-3',5'-cyclic-monophosphate. Similar results were obtained with the electrical cell-substrate impedance sensor (ECIS) technique. Addition of NO donors or L-arginine, but not 8-bromoguanosine-3',5'-cyclic- monophosphate or N2,2'-O-dibutyrylguanosine-3',5'-cyclic-monophosphate, to confluent PMVEC monolayers resulted in a transient decrease in cell adhesion, which was quantitated by the ECIS. Exposure to 1 U/ml α-thrombin reduced the monolayer electrical resistance by ~50%. The observed response was significantly suppressed by pretreatment of cells with intracellular calcium chelator, 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid or NO synthase inhibitor, N(G)-nitro-L-arginine methyl ester, but not guanylate cyclase inhibitor, 6-anilino-5,8-quinoline-quinone. Selective knockout of endothelial NO synthase with antisense oligodeoxynucleotides also significantly reduced thrombin-induced decrease in monolayer resistance. Our findings indicate that thrombin stimulates calcium-dependent release of NO from PMVEC, which mediates the retraction of endothelial cells via a cGMP- independent pathway. Our results suggest that NO modulates cell-matrix and/or cell-cell adhesion in PMVEC and that this molecule might modify microvascular permeability in the human lung. (C) 2000 Elsevier Science Inc.
KW - Cell adhesion
KW - Electrical cell-substrate impedance sensor
KW - Human
KW - Nitric oxide
KW - Pulmonary microvascular endothelial cell
UR - http://www.scopus.com/inward/record.url?scp=0034717271&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034717271&partnerID=8YFLogxK
U2 - 10.1016/S0024-3205(00)00598-1
DO - 10.1016/S0024-3205(00)00598-1
M3 - Article
C2 - 10896023
AN - SCOPUS:0034717271
VL - 67
SP - 1
EP - 11
JO - Life Sciences
JF - Life Sciences
SN - 0024-3205
IS - 1
ER -