克山病患者GPX1-mRNA及凋亡相关信号分子的表达

目的 检测克山病(KSD)患者谷胱甘肽过氧化物酶(GPX)1 mRNA及凋亡相关信号分子NFКBp65和p53的表达情况.方法 于2009年9月15日在克山病病区陕西省黄陵县店头镇,根据心电图、胸部X片及临床检查,选择16例慢型克山病患者为KSD组,在西安交通大学附属第二医院选择23例正常体检者为对照组,采集空腹肘静脉血5 ml,用Trizol提取总mRNA及蛋白.采用GPX检测试剂盒检测全血GPX活性,采用SYBR Real-Time PCR试剂盒检测GPX1-mRNA表达,采用Western blot法检测凋亡相关信号分子NFкBp65及p53表达.结果 KSD组的GPX活性[(108.61±14.10)U]低于对照组[(122.78±11.89)U,t=2.874,P<0.05].GPX1-mRNA表达水平(0.553±0.299)低于对照组(1.00±0,t=4.964,P<0.01).KSD组NFкBp65表达(0.802±0.057)低于对照组[(1.065±0.355),t=6.829,P<0.01],p53表达(1.604±0.191)高于对照组[(1.137±0.186),t=3.033,P<0.05].结论 KSD患者GPX1-mRNA的表达水平降低可能导致GPX活下降,使细胞内氧化损伤增强,进而激活细胞内凋亡相关信号通路引发心肌细胞凋亡.

Expression of GPX1-mRNA and apoptosis related signal molecular in Keshan disease patients

Objective To investigate the meaning of expression of apoptosis related molecules NFKBp65 and p53 and GPX1-mRNA in patients with Keshan disease(KSD).Methods Sixteen chronic Keshan Disease patients were enrolled in KSD group according to electrocardiogram,chest X ray film and clinical examinations on 15,September in 2009,and 23 healthy people were included in control group from physical examination taken in The Second Affiliated Hospital of Xi'an Jiaotong University.Fresh blood(5 ml)was collected from antecubital vein of all subjects in the fasting state.Total mRNA and protein of blood sample were isolated using Trizol.GPX Assay Kit was used to detect GPX enzyme activity,and GPX1-mRNA expression was determined by SYBR Real-Time PCR.Meanwhile,expression of apoptosis related molecules NFKBp65 and p53 were determined by Western blot.Results GPX enzyme activity decreased significantly in KSD group[(108.61±14.10)U]compared with control group[(122.78±11.89)U,t=2.874,P<0.05],GPX1-mRNA level of KSD group(0.553±0.299)notably KSD group(0.802±0.057)compared with control group[(1.065±0.355),t=6.829,P<0.01].p53 increased in KSD group(1.604±0.191)compared with control group[(1.137±0.186),t=3.033,P<0.05].Conclusiom Decreased GPX1-mRNA expression may result in lower GPX enzyme activity of patients with KSD.Thus oxidative damage increases and cadioeyte apoptosis is activated by activating apoptosis signal pathway.