摘要
目的:GPR54是下丘脑神经元调控下丘脑-垂体-性腺轴功能的门控,本文研究不同浓度脂多糖(LPS)、肿瘤坏死因子α(TNFα)、白细胞介素6(IL-6)、地塞米松和胰岛素对人乳腺癌MCF7细胞GPR54 mRNA及蛋白表达的影响。方法:培养MCF7细胞,用LPS(10μg/ml和20μg/ml)、TNFα(20 ng/ml和100 ng/ml)、IL-6(10 ng/ml和20 ng/ml)、地塞米松(10-6mol/L和10-7mol/L)和胰岛素(0.01 IU/L和0.1 IU/L)干预72 h,评价干预6、24、48、72 h后GPR54 mRNA(实时荧光定量PCR)和蛋白表达量(Western印迹)的变化。结果:和对照组相比,LPS(10μg/ml和20μg/ml)、TNFα(20 ng/ml和100 ng/ml)、IL-6(10 ng/ml和20 ng/ml)、地塞米松(10-6mol/L和10-7mol/L)和胰岛素(0.01 IU/L和0.1 IU/L)均显著增加GPR54 mRNA(P均<0.05)和蛋白表达。结论:LPS、TNFα、IL-6、地塞米松和胰岛素显著增加MCF7细胞GPR54的表达,提示这些炎症因子和激素可能通过调节GPR54水平变化进而影响性腺轴功能。
Objective： To investigate the effects of different concentrations of lipopolysaccharide（ LPS）,tumor necrosis factorα（ TNFα）,interleukin-6（ IL-6）,dexamethasone（ Dex）,and insulin on the mRNA and protein expressions of GPR54 in the MCF7 cell line in vitro. Methods： MCF7 breasr cancer cells were cultured and treated with different concentrations of LPS（ 10 and20 μg / ml）,TNFα（ 20 and 100 ng / ml）,IL-6（ 10 and 20 ng / ml）,Dex（ 10^- 6and 10^- 7mol / L）,and insulin（ 0. 01 and0. 1 IU / L）. Those treated with culture fluid only served as controls. The mRNA and protein expressions of GPR54 were measured by real-time PCR and Western blot,respectively,after 6,24,48,and 72 hours of treatment. Results： Compared with the blank control,LPS（ 10 and 20 μg / ml）,TNFα（ 20 and 100 ng / ml）,IL-6（ 10 and 20 ng / ml）,Dex（ 10^- 6and 10^- 7mol / L）,and insulin（ 0. 01 and 0. 1 IU/L） significantly increased the expressions of GPR54 mRNA（ P〈0. 05） and protein（ P〈0. 05）. Conclusion：LPS,TNFα,IL-6,Dex,and insulin evidently increase the expression of GPR54 in the MCF7 cell line,indicating their influence on the function of gonads by regulating the GPR54 level.
出处
《中华男科学杂志》
CAS
CSCD
北大核心
2015年第7期587-592,共6页
National Journal of Andrology
基金
国家自然科学基金青年基金(81100416)~~
