He Xi Jun, Ejiri Noriko, Nakayama Hiroyuki, Doi Kunio
Department of Veterinary Pathology, Graduate School of Agricultural and Life Sciences, The University of Yayoi 1-1-1, Bunkyo-ku, Tokyo, Japan.
Exp Mol Pathol. 2005 Feb;78(1):64-70. doi: 10.1016/j.yexmp.2004.08.011.
A body of evidence suggests that pregnancy may be responsible for the depression in the microsomal enzyme activity and the reduction in the total content of cytochrome P450 (CYP) in the rat liver. However, changes in expression of individual CYP isozyme remain poorly known. The current study was designed to examine the changes in CYPs protein expression in the liver of F344 rats in midpregnancy and late pregnancy by Western blot analysis and immunohistochemistry. Total nine antirat CYPs antibodies (CYP1A1, CYP2B1/CYP2B2, CYP2C6, CYP2C12, CYP2D1, CYP2D4, CYP2E1, CYP3A1, and CYP4A1) were used. In comparison with age-matched nonpregnant control rats, there were significant decreases in hepatic levels of CYP2B2, CYP2C6, and CYP4A1 in midpregnancy (day 13) and CYP2B2, CYP2C6, CYP4A1, CYP1A1, CYP2B1, and CYP2E1 in late pregnancy (day 19). The expression of CYP2C12, CYP2D1, and CYP 3A1 did not differ between nonpregnant and pregnant rats, and CYP2D4 was not detectable in microsomal proteins obtained from nonpregnant and pregnant rats at a protein loading of 20 mug total protein per lane. Immunohistochemistry showed that there were no differences in the distribution and degree of immunostainability for the abovementioned antibodies to nine CYPs between pregnant and nonpregnant rats.
大量证据表明,妊娠可能是大鼠肝脏微粒体酶活性降低及细胞色素P450(CYP)总含量减少的原因。然而,个体CYP同工酶表达的变化仍知之甚少。本研究旨在通过蛋白质印迹分析和免疫组织化学检测妊娠中期和晚期F344大鼠肝脏中CYPs蛋白表达的变化。共使用了九种抗大鼠CYPs抗体(CYP1A1、CYP2B1/CYP2B2、CYP2C6、CYP2C12、CYP2D1、CYP2D4、CYP2E1、CYP3A1和CYP4A1)。与年龄匹配的未孕对照大鼠相比,妊娠中期(第13天)CYP2B2、CYP2C6和CYP4A1的肝脏水平显著降低,妊娠晚期(第19天)CYP2B2、CYP2C6、CYP4A1、CYP1A1、CYP2B1和CYP2E1的肝脏水平显著降低。未孕和妊娠大鼠之间CYP2C12、CYP2D1和CYP 3A1的表达没有差异,在每泳道总蛋白上样量为20μg时,未孕和妊娠大鼠微粒体蛋白中均未检测到CYP2D4。免疫组织化学显示,妊娠和未孕大鼠之间,上述针对九种CYPs的抗体的分布和免疫染色程度没有差异。