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不同浓度ox-LDL对大鼠原代卵泡膜细胞增殖及激素合成相关基因LXR-α和StAR表达的影响 预览

Effects of different concentrations of ox-LDL on the proliferation of rat theca cells and the expression of steroidogenesis related genes LXR-α and StAR
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摘要 目的·初步探讨ox-LDL对大鼠卵泡膜细胞增殖和雄激素合成相关基因LXR-α和St AR表达的影响。方法·免疫组化检测大鼠卵巢组织中LXR-α的表达。体外分离培养原代大鼠卵泡膜细胞,分别用25、50、100、150、200、300和400 mg/L的ox-LDL处理,用real-time PCR检测LXR-αm RNA的变化,用MTT检测细胞活力,用Western blotting检测LXR-α和St AR蛋白的表达。结果·ox-LDL对大鼠卵泡膜细胞增殖的影响和对LXR-α和St AR表达的调控呈现浓度依赖性变化。ox-LDL刺激24 h后,低浓度的ox-LDL(25-150 mg/L)可诱导卵泡膜细胞增殖,以100 mg/L的ox-LDL对卵泡膜细胞增殖的促进作用显著。而当ox-LDL浓度继续上升,细胞存活率下降,以400 mg/L的ox-LDL对卵泡膜细胞增殖的抑制作用显著。低浓度ox-LDL(25-150 mg/L)刺激,导致LXR-αm RNA的表达升高,其中150 mg/L ox-LDL对LXR-αm RNA的表达量影响显著。高浓度ox-LDL抑制LXR-αm RNA的表达,其中400 mg/L的ox-LDL对LXR-αm RNA的表达量影响显著。150 mg/L ox-LDL促进大鼠卵泡膜细胞LXR-α和St AR蛋白的表达上升,但150 mg/L ox-LDL对St AR蛋白表达的促进作用,与对照组相比,差异没有统计学意义;而400 mg/L ox-LDL能显著抑制大鼠卵泡膜细胞LXR-α和St AR蛋白的表达。结论·低浓度的ox-LDL可诱导卵泡膜细胞增殖,促进LXR-α和St AR的表达,而高浓度oxLDL降低细胞活力,抑制LXR-α和St AR的表达。 Objective · To investigate the effects of ox-LDL on the proliferation of rat theca cells and expression of LXR-α and StAR, two genes associated with androgen biosynthesis. Methods · The expression of LXR-α in the ovarian tissue of rats was determined by immunohistochemistry. Primary theca cells were isolated and collected from rat ovary and cultured in vitro. Furthermore, the theca cells were treated with 25, 50, 100, 150, 200, 300 and 400 mg/L ox-LDL,respectively. The variations in LXR-α mRNA were identified using real-time PCR. MTT assay was performed to detect cell viability. The expression of LXR-α and StAR was measured by Western blotting analysis. Results · The effect of ox-LDL on the proliferation of rat theca cells and the levels of LXR-α and StAR in theca cells was in a concentration-dependent manner. Following exposure to various concentration of ox-LDL for 24 h, the proliferation of theca cells was induced by low concentration of ox-LDL (25-150 mg/L), and 100 mg/L ox-LDL showed the most significant inducing effect. Moreover, the cell survival rate was diminished considerably following with ox-LDL concentration increasing, especially lowered by 400 mg/L ox-LDL. The mRNA level of LXR-α was increased with low concentration of ox-LDL (25-150 mg/L) and the impact of ox-LDL on the induced expression of LXR-α mRNA was considerably distinct at the concentration of 150 mg/L. On the other hand, the expression of LXR-α mRNA was reduced with high concentration of ox-LDL, and the impact of 400 mg/L ox-LDLwas substantially distinct. The protein expression levels of LXR-α and StAR were increased with 150 mg/L ox-LDL, but StAR protein level in 150 mg/L ox-LDL group revealed no significant difference when compared with control group. The expression of LXR-α and StAR protein was significantly inhibited with 400 mg/L ox-LDL in the rat theca cells. Conclusion · Low concentrations of ox-LDL can induce the proliferation of theca cells, and promote the expression of StAR and LXR-α. Whereas, high con
作者 陈莹 张益 李聪 CHEN Ying1, ZHANG Yi2, LI Cong1 (1. Department of Obstetrics and Gynecology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400042, China; 2. Key Laboratory of Birth Defects and Reproductive Health of National Health and Family Planning Commission, Chongqing Population and Family Planning Science and Technology Research Institute, Chongqing 400020, China)
出处 《上海交通大学学报:医学版》 CAS CSCD 北大核心 2017年第3期318-324,共7页 Journal of Shanghai Jiaotong University:Medical Science
基金 重庆市卫生和计划生育委员会2011年医学科研计划项目(2011-2-104)
关键词 卵泡膜细胞 氧化型低密度脂蛋白 增殖 肝X受体 类固醇激素合成急性调节蛋白 theca cells oxidized low density lipoprotein proliferation liver X receptor StAR
作者简介 陈莹(1980-),女,讲师,博士。电子信箱:chenying9851231@163.com。
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