Regulation of potassium channels in coronary smooth muscle by adenoviral expression of cytochrome P-450 epoxygenase

被引:21
作者
Campbell, WB
Holmes, BB
Falck, JR
Capdevila, JH
Gauthier, KM
机构
[1] Med Coll Wisconsin, Dept Pharmacol & Toxicol, Milwaukee, WI 53226 USA
[2] Univ Texas, SW Med Ctr, Dept Biochem, Dallas, TX USA
[3] Vanderbilt Univ, Dept Med & Biochem, Nashville, TN USA
来源
AMERICAN JOURNAL OF PHYSIOLOGY-HEART AND CIRCULATORY PHYSIOLOGY | 2006年 / 290卷 / 01期
关键词
endothelium-derived hyperpolarizing factor; arachidonic acid; vascular relaxation;
D O I
10.1152/ajpheart.00516.2005
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Epoxyeicosatrienoic acids (EETs) are endothelium-derived cytochrome P-450 (CYP) metabolites of arachidonic acid that relax vascular smooth muscle by large- conductance calcium- activated potassium (BKCa) channel activation and membrane hyperpolarization. We hypothesized that if smooth muscle cells (SMCs) had the capacity to synthesize EETs, endogenous EET production would increase BKCa channel activity. Bovine coronary SMCs were transduced with adenovirus coding the CYP Bacillus megaterium -3 (F87V) (CYP BM-3) epoxygenase that metabolizes arachidonic acid exclusively to 14(S), 15(R)EET. Adenovirus containing the cytomegalovirus promoter- Escherichia coli beta-galactosidase was used as a control. With the use of an anti-CYP BM-3 (F87V) antibody, a 124-kDa immunoreactive protein was detected only in CYP BM-3-transduced cells. Protein expression increased with increasing amounts of virus. When CYP BM-3-transduced cells were incubated with [C-14] arachidonic acid, HPLC analysis detected 14,15-dihydroxyeicosatrienoic acid (14,15- DHET) and 14,15-EET. The identity of 14,15-EET and 14,15-DHET was confirmed by mass spectrometry. In CYP BM-3-transduced cells, methacholine (10(-5) M) increased 14,15-EET release twofold and BKCa channel activity fourfold in cell- attached patches. Methacholine-induced increases in BKCa channel activity were blocked by the CYP inhibitor 17-octadecynoic acid (10(-5) M). 14 S), 15(R)-EET was more potent than 14(R), 15(S)-EET in relaxing bovine coronary arteries and activating BKCa channels. Thus CYP BM-3 adenoviral transduction confers SMCs with epoxygenase activity. These cells acquire the capacity to respond to the vasodilator agonist by synthesizing 14(S), 15(R)- EET from endogenous arachidonic acid to activate BKCa channels. These studies indicate that 14(S), 15(R)-EET is a sufficient endogenous activator of BKCa channels in coronary SMCs.
引用
收藏
页码:H64 / H71
页数:8
相关论文
共 37 条
  • [1] Comparison of endothelium-dependent relaxation in carotid arteries from Japanese white and Watanabe heritable hyperlipidemic rabbits
    Ayajiki, K
    Ozaki, M
    Shiomi, M
    Okamura, T
    Toda, N
    [J]. JOURNAL OF CARDIOVASCULAR PHARMACOLOGY, 2000, 36 (05) : 622 - 630
  • [2] 14,15-dihydroxyeicosatrienoic acid relaxes bovine coronary arteries by activation of KCa channels
    Campbell, WB
    Deeter, C
    Gauthier, KM
    Ingraham, RH
    Falck, JR
    Li, PL
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY-HEART AND CIRCULATORY PHYSIOLOGY, 2002, 282 (05): : H1656 - H1664
  • [3] Identification of epoxyeicosatrienoic acids as endothelium-derived hyperpolarizing factors
    Campbell, WB
    Gebremedhin, D
    Pratt, PF
    Harder, DR
    [J]. CIRCULATION RESEARCH, 1996, 78 (03) : 415 - 423
  • [4] Biochemical and molecular properties of the cytochrome P450 arachidonic acid monooxygenases
    Capdevila, JH
    Falck, JR
    [J]. PROSTAGLANDINS & OTHER LIPID MEDIATORS, 2002, 68-9 : 325 - 344
  • [5] Transfection of an active cytochrome P450 arachidonic acid epoxygenase indicates that 14,15-epoxyeicosatrienoic acid functions as an intracellular second messenger in response to epidermal growth factor
    Chen, JK
    Wang, DW
    Falck, JR
    Capdevila, J
    Harris, RC
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (08) : 4764 - 4769
  • [6] Comparison of vasodilatory properties of 14,15-EET analogs: structural requirements for dilation
    Falck, JR
    Krishna, UM
    Reddy, YK
    Kumar, PS
    Reddy, KM
    Hittner, SB
    Deeter, C
    Sharma, KK
    Gauthier, KM
    Campbell, WB
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY-HEART AND CIRCULATORY PHYSIOLOGY, 2003, 284 (01): : H337 - H349
  • [7] Cytochrome P4502C is an EDHF synthase in coronary arteries
    Fisslthaler, B
    Popp, R
    Kiss, L
    Potente, M
    Harder, DR
    Fleming, I
    Busse, R
    [J]. NATURE, 1999, 401 (6752) : 493 - 497
  • [8] Cytochrome P450 enzymes in vascular homeostasis
    Fleming, I
    [J]. CIRCULATION RESEARCH, 2001, 89 (09) : 753 - 762
  • [9] DECREASED ENDOTHELIUM-DEPENDENT HYPERPOLARIZATION TO ACETYLCHOLINE IN SMOOTH-MUSCLE OF THE MESENTERIC-ARTERY OF SPONTANEOUSLY HYPERTENSIVE RATS
    FUJII, K
    TOMINAGA, M
    OHMORI, S
    KOBAYASHI, K
    KOGA, T
    TAKATA, Y
    FUJISHIMA, M
    [J]. CIRCULATION RESEARCH, 1992, 70 (04) : 660 - 669
  • [10] Alterations in endothelium-dependent hyperpolarization and relaxation in mesenteric arteries from streptozotocin-induced diabetic rats
    Fukao, M
    Hattori, Y
    Kanno, M
    Sakuma, I
    Kitabatake, A
    [J]. BRITISH JOURNAL OF PHARMACOLOGY, 1997, 121 (07) : 1383 - 1391