孕马血清促性腺激素对小鼠肝脏组织CYP17A1和PXR表达的影响

Effect of PMSG on Expressions of CYP17A1 and PXR in Mouse Liver Tissue

旨在了解孕马血清促性腺激素(pregnant mare serum gonadotropin,PMSG)对小鼠肝脏细胞色素P45017A1(cytochrome P45017A1,CYP17A1)和孕烷X受体(pregnane X receptor,PXR)表达的影响。将48只雌性昆明系小鼠随机分为PMSG处理组和对照组,PMSG组小鼠腹腔注射PMSG 10 IU,注射后12、24、48 h处死12只小鼠,采血分离血清并采集肝脏组织样本;对照组腹腔注射等体积生理盐水,在注射后12、24、48 h时采集4只小鼠的血液和肝脏组织样本。酶联免疫吸附试验(ELISA)检测血清PMSG含量,苏木精-伊红染色(HE)观察PMSG对肝脏组织的影响,RT-qPCR和Western blot检测肝脏组织CYP17A1和PXR表达水平,免疫组织化学染色检测CYP17A1和PXR在肝脏组织中的定位。结果显示,PMSG注射后12 h小鼠血清中PMSG含量显著升高(P<0.05),24 h和48 h时逐渐降低,但仍显著高于NC组(P<0.05);PMSG注射后12、24、48 h时,肝脏细胞水泡变性明显;注射PMSG后肝脏组织中CYP17A1和PXR基因和蛋白表达均显著下调(P<0.05);CYP17A1和PXR主要定位于肝索肝实质细胞质中。研究结果证明,PMSG能够显著抑制肝脏CYP17A1和PXR表达。

The purpose of this study was to investigate the effect of pregnant mare serum gonadotropin(PMSG)on the expression of cytochrome P45017A1(CYP17A1)and pregnane X receptor(PXR)in mouse liver.48 Kunming female mice were randomly divided into PMSG treatment group(PMSG group)and control group.The mice in PMSG group were intraperitoneally injected with PMSG 10 IU.At 12,24 and 48 hours after injection,12 mice were killed,blood samples were collected,and liver tissue samples were collected;The mice in control group were intraperitoneally injected with the same volume of normal saline.At 12 h,24 h and 48 h after injection,blood and liver tissue samples were collected from 4 mice.The serum PMSG levels were detected by enzyme-linked immunosorbent assay(ELISA);hematoxylin-eosin staining(HE)was used to observe the effect of PMSG on liver;RT-qPCR and Western blot were used to detect the expression levels of CYP17A1 and PXR in liver;immunohistochemistry was used to detect the localization of CYP17A1 and PXR in liver tissue.The results showed that the content of PMSG in serum of the mice was significantly increased at 12 h after PMSG injection(P<0.05),and gradually decreased at 24 h and 48 h,but still significantly higher than that in NC group(P<0.05);At 12 h,24 h and 48 h after PMSG injection,the liver cells showed obvious vesicular degeneration;The expressions of CYP17A1 and PXR genes and proteins in liver tissues were significantly decreased after PMSG injection(P<0.05);CYP17A1 and PXR were mainly located in the cytoplasm of hepatic cord and liver parenchyma.The results showed that PMSG could significantly inhibit the expressions of CYP17Al and PXR in liver.