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Estrogen‐dependent upregulation of CYP 2C family in arteries of eNOS deficient mice
Author(s) -
Huang An,
Jacobson Azita,
Ojaimi Caroline,
Sun Dong,
Kaley Gabor
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.a1412-b
Subject(s) - enos , downregulation and upregulation , medicine , endocrinology , estrogen , western blot , cytochrome p450 , microarray analysis techniques , biology , nitric oxide synthase , chemistry , gene expression , nitric oxide , gene , biochemistry , metabolism
Our previous studies demonstrated that epoxyeicosatrienoic acids (EETs), metabolites of arachidonic acid via a cytochrome P450 (CYP) pathway, are endothelium‐derived hyperpolarizing factors (EDHF) in microvessels of female mice and rats, as a function of NO deficiency. In the present studies, we provided molecular evidence of upregulation of CYP 2C family, responsible for the synthesis of EETs, in isolated arteries/arterioles of female endothelial nitric oxide knockout (eNOS‐KO) mice by microarray. Mesenteric arterioles of both genders of eNOS‐KO and wild type (WT) mice were isolated and snap frozen in liquid nitrogen. ~10 μg of total RNA were isolated, and a total of 10 arrays from 2 WT and 3 eNOS‐KO mice of each gender were performed. Focusing on CYP gene expression, microarray results indicate a specific upregulation of CYP 2C29/38/40 (>2‐fold, P<0.05) in female eNOS‐KO vessels compared to male eNOS‐KO vessels. The significant difference was further confirmed by qRT‐PCR and Western blot analysis. There is no significant difference in the expression of CYP2C29/38/40 between the vessels of WT male and female mice, indicating that a co‐existence of both the presence of estrogen and NO deficiency is required for the upregulation of these genes. Moreover, CYP 2C29 expression in vessels of L‐NAME‐treated WT female mice was also determined by RT‐PCR, indicating a significant increase in CYP 2C29 mRNA compared to that of L‐NAME‐treated WT male mice. In summary, the CYP 2C family, especially 2C29/38/40, is the most likely candidate for the CYP metabolite‐mediated vasodilator/hyperpolarizing responses to shear stress in microvessels of female NO deficient mice and rats. (Supported by NIH HL‐43023, HL‐68813 and HL‐070653).