苦荞麦总黄酮对胰岛素抵抗状态EA.hy926细胞ADMA/PRMTⅠ/eNOS/NO的影响  

The effect of total flavonoids of tartary buckwheat on ADMA/PRMTⅠ/eNOS/NO on insulin resistance in EA.hy926 cells

作　　者：赵丹玉[1] 柳春[1] 李宝坤[1] 冯晓帆[1] 张林[1] 

机构地区：[1]辽宁中医药大学基础医学院,辽宁沈阳110847

出　　处：《沈阳药科大学学报》2016年第6期470-474,479,共6页Journal of Shenyang Pharmaceutical University

基　　金：辽宁省杰出青年学者成长计划资助项目(LJQ2013102);辽宁省博士科研启动基金资助项目(20121100);辽宁省社发攻关及成果产业化资助项目(2013020195-202)

摘　　要：目的研究苦荞麦总黄酮对胰岛素抵抗状态EA.hy926细胞内NO、不对称二甲基精氨酸(asymmetric dimethyl arginine,ADMA)的合成及相关基因PRMTⅠ、e NOS表达的影响。方法将培养的EA.hy926细胞分为正常对照组、模型组、苦荞麦总黄酮组及二甲双胍组。采用硝酸还原酶法测定细胞培养基中NO的含量;采用双抗体夹心法测定细胞中ADMA的含量,采用RT-PCR以及免疫印迹(western blot)法分别测定各组细胞e NOS及PRMTⅠmRNA和蛋白的表达。结果同正常组相比,模型组细胞培养基中NO含量明显降低(P<0.01),细胞中ADMA含量显著升高(P<0.01),e NOS mRNA及蛋白表达明显降低(P<0.01),PRMTⅠmRNA和蛋白的表达显著升高(P<0.01);同模型组相比,苦荞麦总黄酮组及二甲双胍组以上指标均有明显改善,两组之间无明显差异。结论苦荞麦总黄酮通过降低胰岛素抵抗状态EA.hy926细胞PRMTⅠmRNA及蛋白的表达,降低细胞中ADMA的含量,提高eNOS mRNA及蛋白表达,使NO合成增多,改善胰岛素抵抗。Objective To study the effect of total flavonoids of tartary buckwheat on ADMA,PRMT Ⅰ,e NOS and NO on insulin resistance in EA. hy926 cells. Methods EA. hy 926 cells were cultured and divided into control group,model group,total flavonoids of tartary buckwheat group and metformin group randomly.The levels of NO in cell culture medium were measured by nitrate reductase test,the contents of ADMA in cells were measured by double antibody,the expressions of e NOS and PRMT Ⅰ mRNA and protein were determined by RT-PCR and western blot respectively. Results compared with control,the levels of NO in model were significantly reduced（ P〈0. 01）,the contents of ADMA in model were significantly higher（ P〈0. 01）,the expression of e NOS mRNA and protein were significantly decreased（ P〈0. 01）,and the expression of PRMT ⅠmRNA and protein were decreased in model（ P〈0. 01）. The above indicators showed the opposite expression in both total flavonoids of tartary buckwheat group and metformin group compared with model（ P〈0. 01）,and there was no significant difference between two groups（ P〈0. 01）. Conclusions Total flavonoids of tartary buckwheat can improve insulin resistance by reducing the expression of PRMT,the content of ADMA,and increasing the expression of e NOS,the synthesis of NO.

关 键 词：苦荞麦总黄酮 胰岛素抵抗 一氧化氮 内皮型一氧化氮合酶 不对称二甲基精氨酸 蛋白精氨酸N-甲基转移酶Ⅰ 

分 类 号：R28[医药卫生—中药学]