| 高效液相色谱法检测心肌细胞蛋白降解率 |
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| 引用本文： | 陈保林,马跃东,熊肇军,张成喜,刘晨,董吁钢.高效液相色谱法检测心肌细胞蛋白降解率[J].重庆医学,2011,40(22):2234-2235,2290. |
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| 作者姓名： | 陈保林 马跃东 熊肇军 张成喜 刘晨 董吁钢 |
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| 作者单位： | 陈保林,Chen Baolin(贵州省人民医院心内科,贵阳,550002);马跃东,Ma Yuedong(中山大学附属第一医院心血管医学部,广州,510080);熊肇军,张成喜,Xiong Zhaojun,Zhang Chengxi(中山大学附属第三医院心内科,广州,510630);刘晨,董吁钢,Liu Chen,Dong Yugang(中山大学附属第一医院心血管医学部,广州,510080;卫生部辅助循环重点实验室,广州,510080) |
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| 摘 要： | 目的建立一种快速测定心肌细胞3-甲基组氨酸(3-MH)释放量的方法。方法分离培养新生大鼠心肌细胞。细胞培养液样品用乙腈沉淀蛋白,离心后取上清液进样用于3-MH分析。3-MH检测采用岛津LC-20A高效液相色谱仪,用C18色谱柱分离,外标法定量,检测波长为338 nm。结果色谱峰分离良好,线性方程为:y=731 241x+896.86,相关系数(r)=0.999 8,3-MH线性检测范围为1.6～107.0μmol/L。结论该实验为检测心肌细胞蛋白降解提供了一种精确、简单、可行的方法。
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| 关 键 词： | 肌细胞 心脏 色谱法 高压液相 甲基组氨酸类 |
Determination of protein degradation in cardiomyocytes by high performance liquid chromatography |
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| Chen Baolin,Ma Yuedong,Xiong Zhaojun,Zhang Chengxi,Liu Chen,Dong Yugang.Determination of protein degradation in cardiomyocytes by high performance liquid chromatography[J].Chongqing Medical Journal,2011,40(22):2234-2235,2290. |
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| Authors: | Chen Baolin Ma Yuedong Xiong Zhaojun Zhang Chengxi Liu Chen Dong Yugang |
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| Institution: | 2,4(1.Department of Cardiology,the People’s Hospital of Guizhou Province,Guiyang 550002,China;2.Department of Cardiology,the First Affiliated hospital of Sun Yat-Sen University,Guangzhou 510080,China;3.Department of Cardiology,the Third Affiliated Hospital of Sun Yat-Sen University,Guangzhou 510630,China;4.Key Laboratory on Assisted Circulation of Ministry of Health,Guangzhou 510630,China) |
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| Abstract: | Objective To establish a fast method to determine the release of 3-methylhistidine(3-MH) in cardiomyocytes.Methods Neonatal SD rat cardiomyocytes were isolated and cultured.The analytes in culture medium were extracted by protein precipitation using acetonitrile.High performance liquid chromatographic detection was used to determine the 3-MH levels in medium by applying Shimadzu High-Performance Liquid Chromatograph.Chromatographic separation was performed on a C18 column.The monitor wave-length was 338 nm.Results The chromatography was well separative.Linear equation was:y=731 241X+896.86,r=0.999 8.The calibration curve was linear in the range of 1.6-107.0 μmo/L for 3-MH,with correlation coefficient above 0.99.Conclusion The method is accurate,precise and reliable for the determination of protein degradation in cardiomyocytes. |
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| Keywords: | myocytes cardiac chromatography high pressure liquid methylhistidines |
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