Abstract
Epoxyeicosatrienoic acids (EETs), cytochrome P450-derived metabolites of arachidonic acid, have been reported to increase intracellular calcium concentration in aortic vascular smooth muscle cells (SMCs). As EETs are labile, we synthesized a new stable urea EET analog with agonist and soluble epoxide hydrolase (sEH) inhibitor properties. We refer to this analog, 12-(3-hexylureido)dodec-8-enoic acid, as 8-HUDE. Measuring tension of vascular rings, intracellular calcium signaling by confocal laser scanning microscopy and gene expression by reverse-transcription-PCR and western blots, we examined the effects of 8-HUDE on pulmonary vascular tone and calcium signaling in rat pulmonary artery (PA) SMCs (PASMCs). 8-HUDE increased the tension of rat PAs to 145% baseline, whereas it had no effect on the tension of mesenteric arteries (MAs). The 8-HUDE-induced increase in vascular tone was abolished by removal of extracellular Ca2+ or by pretreatment with either La 3 or SKF96365, which are inhibitors of canonical transient receptor potential channels (TRPCs). Furthermore, 8-HUDE-evoked increases in Ca2+ i in PASMCs could be blunted by inhibition of TRPC with SKF96365, removal of extracellular calcium or depletion of intracellular calcium stores with caffeine, cyclopiazonic acid or 2-aminoethoxydiphenyl borate, but not by the voltage-activated calcium channel blocker nifedipine. In addition to immediate effects on calcium signaling, 8-HUDE upregulated the expression of TRPC1 and TRPC6 at both mRNA and protein levels in rat PASMCs, whereas it suppressed the expression of sEH. Our observations suggest that 8-HUDE increases PA vascular tone through increased release of calcium from intracellular stores, enhanced Ca2+ i influx in PASMCs through store-operated Ca2+ channels and modulated the expression of TRPC and sEH proteins in a proconstrictive manner.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 630-639 |
| Number of pages | 10 |
| Journal | Hypertension Research |
| Volume | 34 |
| Issue number | 5 |
| DOIs | |
| State | Published - May 2011 |
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Keywords
- 12-(3-hexylureido)dodec-8-enoic acid
- 8-HUDE
- canonical transient receptor potential channel (TRPC)
- epoxyeicosatrienoic acids (EETs)
- pulmonary arteries
- store-operated Ca channels (SOCCs)
ASJC Scopus subject areas
- Internal Medicine
- Physiology
- Cardiology and Cardiovascular Medicine
Cite this
Stable EET urea agonist and soluble epoxide hydrolase inhibitor regulate rat pulmonary arteries through TRPCs. / Liu, Yun; Wang, Ruifang; Li, Jing; Rao, Jingjing; Li, Weiyang; Falck, J R; Manthati, Vijay L.; Medhora, Meetha; Jacobs, Elizabeth R.; Zhu, Daling.
In: Hypertension Research, Vol. 34, No. 5, 05.2011, p. 630-639.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Stable EET urea agonist and soluble epoxide hydrolase inhibitor regulate rat pulmonary arteries through TRPCs
AU - Liu, Yun
AU - Wang, Ruifang
AU - Li, Jing
AU - Rao, Jingjing
AU - Li, Weiyang
AU - Falck, J R
AU - Manthati, Vijay L.
AU - Medhora, Meetha
AU - Jacobs, Elizabeth R.
AU - Zhu, Daling
PY - 2011/5
Y1 - 2011/5
N2 - Epoxyeicosatrienoic acids (EETs), cytochrome P450-derived metabolites of arachidonic acid, have been reported to increase intracellular calcium concentration in aortic vascular smooth muscle cells (SMCs). As EETs are labile, we synthesized a new stable urea EET analog with agonist and soluble epoxide hydrolase (sEH) inhibitor properties. We refer to this analog, 12-(3-hexylureido)dodec-8-enoic acid, as 8-HUDE. Measuring tension of vascular rings, intracellular calcium signaling by confocal laser scanning microscopy and gene expression by reverse-transcription-PCR and western blots, we examined the effects of 8-HUDE on pulmonary vascular tone and calcium signaling in rat pulmonary artery (PA) SMCs (PASMCs). 8-HUDE increased the tension of rat PAs to 145% baseline, whereas it had no effect on the tension of mesenteric arteries (MAs). The 8-HUDE-induced increase in vascular tone was abolished by removal of extracellular Ca2+ or by pretreatment with either La 3 or SKF96365, which are inhibitors of canonical transient receptor potential channels (TRPCs). Furthermore, 8-HUDE-evoked increases in Ca2+ i in PASMCs could be blunted by inhibition of TRPC with SKF96365, removal of extracellular calcium or depletion of intracellular calcium stores with caffeine, cyclopiazonic acid or 2-aminoethoxydiphenyl borate, but not by the voltage-activated calcium channel blocker nifedipine. In addition to immediate effects on calcium signaling, 8-HUDE upregulated the expression of TRPC1 and TRPC6 at both mRNA and protein levels in rat PASMCs, whereas it suppressed the expression of sEH. Our observations suggest that 8-HUDE increases PA vascular tone through increased release of calcium from intracellular stores, enhanced Ca2+ i influx in PASMCs through store-operated Ca2+ channels and modulated the expression of TRPC and sEH proteins in a proconstrictive manner.
AB - Epoxyeicosatrienoic acids (EETs), cytochrome P450-derived metabolites of arachidonic acid, have been reported to increase intracellular calcium concentration in aortic vascular smooth muscle cells (SMCs). As EETs are labile, we synthesized a new stable urea EET analog with agonist and soluble epoxide hydrolase (sEH) inhibitor properties. We refer to this analog, 12-(3-hexylureido)dodec-8-enoic acid, as 8-HUDE. Measuring tension of vascular rings, intracellular calcium signaling by confocal laser scanning microscopy and gene expression by reverse-transcription-PCR and western blots, we examined the effects of 8-HUDE on pulmonary vascular tone and calcium signaling in rat pulmonary artery (PA) SMCs (PASMCs). 8-HUDE increased the tension of rat PAs to 145% baseline, whereas it had no effect on the tension of mesenteric arteries (MAs). The 8-HUDE-induced increase in vascular tone was abolished by removal of extracellular Ca2+ or by pretreatment with either La 3 or SKF96365, which are inhibitors of canonical transient receptor potential channels (TRPCs). Furthermore, 8-HUDE-evoked increases in Ca2+ i in PASMCs could be blunted by inhibition of TRPC with SKF96365, removal of extracellular calcium or depletion of intracellular calcium stores with caffeine, cyclopiazonic acid or 2-aminoethoxydiphenyl borate, but not by the voltage-activated calcium channel blocker nifedipine. In addition to immediate effects on calcium signaling, 8-HUDE upregulated the expression of TRPC1 and TRPC6 at both mRNA and protein levels in rat PASMCs, whereas it suppressed the expression of sEH. Our observations suggest that 8-HUDE increases PA vascular tone through increased release of calcium from intracellular stores, enhanced Ca2+ i influx in PASMCs through store-operated Ca2+ channels and modulated the expression of TRPC and sEH proteins in a proconstrictive manner.
KW - 12-(3-hexylureido)dodec-8-enoic acid
KW - 8-HUDE
KW - canonical transient receptor potential channel (TRPC)
KW - epoxyeicosatrienoic acids (EETs)
KW - pulmonary arteries
KW - store-operated Ca channels (SOCCs)
UR - http://www.scopus.com/inward/record.url?scp=79955600318&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79955600318&partnerID=8YFLogxK
U2 - 10.1038/hr.2011.5
DO - 10.1038/hr.2011.5
M3 - Article
C2 - 21307870
AN - SCOPUS:79955600318
VL - 34
SP - 630
EP - 639
JO - Hypertension Research
JF - Hypertension Research
SN - 0916-9636
IS - 5
ER -