摘要
【目的】观察血管紧张素II(Ang II)对体外培养的小鼠足细胞葡萄糖转运蛋白1(GLUT1)的表达及膜转位的影响。【方法】Ang II对GLUT1m RNA和GLUT1蛋白表达的检测将体外培养的足细胞分为4组:对照组(C)、低剂量组(Ang II 10-10mol/L)、中剂量组(Ang II 10-8 mol/L)及高剂量组(Ang II 10-6 mol/L),用RT-PCR、Western blot检测GLUT1m RNA和GLUT1蛋白的表达;Ang II对GLUT1足细胞胞膜分布检测也将足细胞分为4组:C组、胰岛素(Ins)10-6 mol/L组、Ang II 10-6 mol/L+Ins10-6mol/L组及氯沙坦(LO)10-6 mol/L+Ang II 10-6 mol/L+Ins 10-6 mol/L组,Western blot检测GLUT1在胞膜的蛋白表达及间接免疫荧光观察GLUT1在胞膜的分布。【结果】m RNA与蛋白表达分组中,与对照组相比,Ang II低、中、高浓度3组GLUT1m RNA水平分别增加了5.89%、21.46%、30.95%(P皆<0.01),各组之间差别亦有统计学意义(P<0.01);蛋白表达水平分别增加了8.40%、11.79%、20.57%(P皆<0.01),各组之间差别亦有统计学意义(P<0.01),Spearman相关分析显示Ang II浓度与GLUT1蛋白表达呈正相关(r=0.934,P<0.001)。足细胞胞膜分布检测中,与对照组相比,足细胞胞膜GLUT1蛋白表达Ins 10-6 mol/L组增加了16.33%(P<0.01)、Ang II 10-6 mol/L+Ins 10-6 mol/L组增加了3.27%(P>0.05),LO 10-6 mol/L+Ang II 10-6mol/L+Ins 10-6mol/L组增加了11.33%(P<0.01);免疫荧光显示Ins组GLUT1在胞膜上的分布明显多于对照组及Ang II+Ins组,而LO干预后,胞膜的荧光强度增加。【结论】Ang II能剂量依赖性地增加足细胞胞内GLUT1的表达及合成;Ang II能够阻碍胰岛素诱发的GLUT1向足细胞胞膜的膜转位,氯沙坦可部分阻断Ang II的作用。
【Objective】 To investigate the effect of angiotensin II（Ang II） on the expression and the translocation of Glucose Transporter-1（GLUT-1） in cultured mouse podocyte cells. 【Methods】 Method one, The expression of m RNA and protein of GLUT1：Cultured mouse podocyte cells in vitro were randomly divided into normal control group（C）, Ang II at the concentration of 10^-10 mol / L（low concentration group）,10^-8mol / L（middle concentration group）,10^-6mol / L（high concentration group）. The expression of m RNA and protein of GLUT1 were detected by semi-quantitative RT-PCR and Western blot respectively. Method two, the distribution of GLUT1 in the plasma membrane（translocation） of podocyte： Podocyte Cells were divided into normal control group（C）, insulin10^-6mol / L, Ang II 10^-6mol / L plus insulin 10^-6mol / L and losartan（LO） 10^-6mol / L plus Ang II 10^-6mol / L and insulin 10^-6mol / L groups. The way of Western blot and immunocytofluorescence were used. 【Results】 Compared to control group, Ang II at the concentration of low, middle, and high groups increased GLUT1 transcription by 5.89%（P〈0.01）, 21.46%（P〈0.01）, and30.95%（P〈0.01）, respectively, and increased GLUT1 protein expression by 8.40%（P〈0.01）, 11.79%（P〈0.01）, and 20.57%（P〈0.01）, respectively, and spearman rank correlation discovered positive correlation between Ang II concentration and GLUT1 protein expression（R = 0.934,P〈0.01）. Compared to control group, the amount of GLUT1 in the plasma membrane was increased by 16.33% in insulin group（P〈0.01）, by 3.27% in the Ang II 10^-6mol / L plus insulin 10^-6mol / L group（P〈0.05）, and by11.33% in the LO 10^-6mol / L plus Ang II 10^-6mol / L and insulin 10^-6mol / L group（P〈0.01）. Immunocytofluorescence indicated the expression of GLUT1 in the plasma membrane of insulin group was the strongest in the four groups and losartan could increase the GLUT1 expression in the plasma membrane partly. 【Conclusion】 Ang
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2015年第2期189-194,共6页
Journal of Sun Yat-Sen University:Medical Sciences
基金
广东省自然科学基金(S2011010003081)
