CAJ | 학술논문

目的探讨细胞外信号调节激酶(ERK1/2蛋白)在睾酮对大鼠心肌肥大中的作用.方法用差速贴壁法分离及纯化培养新生SD大鼠心肌细胞,以Bradford法测定心肌细胞蛋白质含量,同位素法分析~3H-亮氨酸(~3H-Leu)掺入,IBAS图像分析心肌细胞表面积,以免疫印迹法检测心肌细胞ERK1/2蛋白表达水平.结果生理浓度的T作用于大鼠心肌细胞24 h后,细胞蛋白质含量、3~H-Leu掺入和细胞表面积均增加,其中以10~(-8) mol/L作用最强.雄激素受体拮抗剂氟他胺(Flu)10~(-5) mol/L预处理2 h可抑制T诱导的心肌细胞蛋白质含虽的增加,而Flu单独作用对心肌细胞蛋白质含量无影响.ERK1/2信号通路的特异性抑制剂PD98059 50μmol/L预处理2 h,可抑制T诱导的心肌细胞~3H-Leu掺入的增加;10~(-8) mol/L T作用24 h使心肌细胞的ERK1/2的蛋白表达显著增加;10~(-5) mol/L Flu预处理2 h可逆转T诱导的心肌细胞ERK1/2蛋白表达的增加.结论生理浓度的T可以诱导心肌细胞肥大反应,该作用可能由ERK1/2信号通路介导.T通过雄激素受体(AR)上调ERK1/2蛋白表达.
목적 탐토세포외신호조절격매(ERK1/2단백)재고동대대서심기비대중적작용.방법 용차속첩벽법분리급순화배양신생SD대서심기세포,이Bradford법측정심기세포단백질함량,동위소법분석~3H-량안산(~3H-Leu)참입,IBAS도상분석심기세포표면적,이면역인적법검측심기세포ERK1/2단백표체수평.결과 생리농도적T작용우대서심기세포24 h후,세포단백질함량、3~H-Leu참입화세포표면적균증가,기중이10~(-8) mol/L작용최강.웅격소수체길항제불타알(Flu)10~(-5) mol/L예처리2 h가억제T유도적심기세포단백질함수적증가,이Flu단독작용대심기세포단백질함량무영향.ERK1/2신호통로적특이성억제제PD98059 50μmol/L예처리2 h,가억제T유도적심기세포~3H-Leu참입적증가;10~(-8) mol/L T작용24 h사심기세포적ERK1/2적단백표체현저증가;10~(-5) mol/L Flu예처리2 h가역전T유도적심기세포ERK1/2단백표체적증가.결론 생리농도적T가이유도심기세포비대반응,해작용가능유ERK1/2신호통로개도.T통과웅격소수체(AR)상조ERK1/2단백표체.
Objective To explore the role of ERK1/2 protein in development of myocardial hypertrophy.Methods Myocardial cells were isolated from ventricles of 1～3-day-old neonate rats and purifed by a culture method.Neonate rat cardiomyocyte hypertrophic responses were assayed by measuring protein content,protein synthesis rate and cell surface area.Expression of protein ERK1/2 were detected by Western blot.Results Cell protein content,~3H-leucine(~3 H-Leu)incorporation and cell surface area increased by treating of cardiomyocytes with T(10~(-10)～10~(-6) mol/L)for 24 h.The maxium effect was observed at the concentration of 10~(-8) mol/L.The increase of cell protein content induced by T was inhibited by pretreating with flutamide(10~(-5) mol/L)for 2 h,while there was no effect on cardiomyocytes pretreating with flutamide alone.The increase of ~3H-Leu incorporation induced by T was blocked by PD98059(50 μmol/L).Expression of ERK1/2 was upregulated significantly by treating with testoster one for 24 h at the level of 10~(-8) mol/L.The increased expression of ERK1/2 induced by T was reversed by pretreating with flutamide(10~(-5) mol/L)for 2 h.Conclusion T with physio-concentration may induce cardiomyocyte hypertrophy and this effect was possibly mediated through the activation of ERK1/2 signalling.During this procession,T upregulated the protein expression of ERK1/2 mediated by androgen receptor.