Sexual dimorphism and tissue specificity in the expression of CYP4F forms in Sprague Dawley rats
Sexual dimorphism and tissue specificity in the expression of CYP4F forms in Sprague Dawley rats
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DOI：
10.1124/dmd.30.9.1022
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发表时间：
2002-09-01
影响因子：
3.9
通讯作者：
Strobel, HW
中科院分区：
文献类型：
--
作者：
Kalsotra, A;Anakk, S;Strobel, HW
细胞色素P450属于参与多种内源性和外源性代谢途径的酶超家族。细胞色素P450（CYP 4F）是一个新的细胞色素P450家族，它特异性地介导白三烯B-4 ω-羟基化，目前已在包括大鼠、小鼠和人类在内的多种物种中鉴定出几个成员。在大鼠中，CYP 4F家族由四个相关基因组成：CYP 4F 1、CYP 4F 4、CYP 4F 5和CYP 4F 6。在这里，我们报告的荧光实时定量聚合酶链反应测定（TaqMan）的发展，使我们能够进行特定的定量mRNA表达的所有四个成员的这个亚家族。由于迄今为止还没有已知的CYP 4F家族诱导剂，我们使用氯贝特（一种已知的大鼠CYP 4F抑制剂）验证了这些试验。此外，进行北方印迹杂交以验证这些测定。使用这种方法，我们证明定量，第一次，每一个大鼠CYP 4Fs的表达在组织和性别依赖性的方式显示出显着更高的表达在女性相对维斯男性。使用CYP 4F多克隆抗体的蛋白质印迹分析也显示，与雄性大鼠相比，雌性大鼠肝脏、肾脏和肺中的蛋白质表达显著升高。此外，我们观察到与对照雌性相比，卵巢切除大鼠肾脏和肝脏中的CYP 4F 1、CYP 4F 4和CYP 4F 6信息显著减少。这两种组织中的表达损失通过雌激素治疗部分恢复，表明雌激素在调节CYP 4F表达中的作用。
The cytochromes P450 belong to a superfamily of enzymes involved in a diverse array of endobiotic and xenobiotic metabolic pathways. Several members of a novel family of cytochrome P450 (CYP4F), which specifically mediate leukotriene B-4 omega-hydroxylation, have now been identified in various species including rat, mouse, and human. In rats, the CYP4F family consists of four related genes, CYP4F1, CYP4F4, CYP4F5, and CYP4F6. Here we report development of fluorescent real-time quantitative polymerase chain reaction assays (TaqMan), which allow us to carry out specific quantitation of mRNA expression of all four members of this subfamily. Since no inducers for the CYP4F family are known to date, we validated these assays using clofibrate, a known suppressor of rat CYP4Fs. Additionally, Northern blot hybridization was carried out to validate these assays. Using this approach, we demonstrate quantitatively, for the first time, that each of the rat CYP4Fs is expressed in a tissue- and sex-dependent manner showing a significantly higher expression in females vis-a-vis males. Western blot analysis using a CYP4F polyclonal antibody also shows a considerably higher protein expression in female liver, kidneys, and lungs when compared with male rats. Furthermore, we observe a significant decrease in the CYP4F1, CYP4F4, and CYP4F6 message in kidneys and liver of ovariectomized rats when compared with control females. This loss of expression is partially restored by estrogen treatment in both tissues, suggesting a role of estrogen in regulating CYP4F expression.