雌激素对乳鼠心肌细胞脑钠肽表达的影响及机制  

Effect of Estrogen on Brain Natriuretic Peptide Expression of Neonatal Rat Cardiomyocytes

作　　者：吴凌凌[1] 陈艺莉[1] 黄艺仪[2] 周越[1] 何建桂[1] 

机构地区：[1]中山大学附属第一医院心内科,广东广州510080 [2]中山大学附属第一医院急诊科,广东广州510080

出　　处：《中山大学学报（医学科学版）》2014年第4期488-493,共6页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(81100171);广东省科技计划项目(2012B031800386)

摘　　要：【目的】探讨雌激素对原代培养的心肌细胞内脑钠肽(BNP)的表达是否有影响以及雌激素影响BNP表达的机制。【方法】视实验目的给予无血清饥饿状态下的心肌细胞不同浓度的雌激素以及相应通路抑制剂处理,以RT-PCR测定细胞内BNP的mRNA水平,以Western-Blotting测定细胞内BNP的蛋白表达量以及相关通路的磷酸化水平,以免疫荧光技术观察雌激素刺激下心肌BNP的亚细胞定位。【结果】雌激素刺激能够显著上调心肌细胞内的BNP mRNA以及蛋白水平(P<0.05),雌激素受体抑制剂ICI 182 780能够抑制该效应(P<0.05)。雌激素处理能够上调心肌Akt以及p38蛋白磷酸化水平(P<0.05),但不能上调Erk1/2蛋白磷酸化水平。预处理PI3K/Akt通路抑制剂LY294002或p38 MAPK通路抑制剂SB203580均能够降低雌激素诱导的心肌细胞BNP表达(P<0.05)。【结论】雌激素能够特异性地通过其受体影响心肌细胞BNP的表达,并且PI3K/Akt通路以及p38 MAPK通路均参与了雌激素诱导的心肌细胞BNP表达过程。[Objective]The purpose of this study was to determine whether estrogen could regulate brain natriuretic peptide （BNP） expression in primary cultured cardiomyocytes and to examine the mechanism that are involved in this process.[Methods]Serum-starved neonatal rat ventricular cardiomyocytes （NRVCs） was treated with different concentrations of 17-beta-estrodiol and related specific signal pathway inhibitors.BNP expression in NRVCs and related signal pathway was analyzed by RT-PCR and Western blotting.The subcellular localization of cardiac BNP under estrogen treatment was determined by immunofluorescence.[Results]Our data showed that estrogen treatment notably increased intracellular BNP expression of NRVCs in a concentration-dependent manner （P 〈 0.05）,which could be eliminated by estrogen receptor antagonist ICI 182 780 （P 〈 0.05）.Estrogen treatment also upregulated the phosphorylation level of Akt and p38 （P 〈 0.05） but not Erk1/2.Furthermore,pretreatment of PI3-kinase/Akt inhibitor LY294002 or p38 MAPK inhibitor SB203580 decreased the estrogen-upregulated BNP expression in NRVCs （P 〈 0.05）.[Conclusion]Estrogen specifically induces intracellular BNP expression in primary cardiomyocytes via estrogen receptor.Both PI3-kinase/Akt and p38 MAPK signal pathways were involved in estrogen-induced BNP expression.

关 键 词：雌激素 BNP AKT P38 心肌细胞 

分 类 号：Q2[生物学—细胞生物学]