成都地区青年人群中血浆和红细胞还原/氧化型谷胱甘肽的含量及其氧化-还原电位的分布

The Measurement of redox state of glutathione in plasma and red blood cell in health youth of chengdu district and their redox potential distribution

目的 :测量谷胱甘肽还原型和氧化型在血浆内和红细胞内的含量 ,探讨GSH/GSSG及其氧化还原电位在健康人群的分布情况。方法 :在组织内谷胱甘肽荧光测量法的基础上 ,加以改进 ,对血浆和红细胞内的GSH/GSSG进行测定 ,并计算出GSH/GSSH的氧化还原电位。同时 ,探讨了冷冻保存和去蛋白处理对血浆和红细胞内GSH和GSSG的影响。结果 :改进后的方法具重复性好 ;准确性高 ,回收率在 98.7%和 97.5 % ;标准曲线呈明显线性等良好的测定效率。用本法对 42名年青年健康志愿者的新鲜血浆和红细胞液的谷胱甘肽进行测定 ,结果为 :血浆内GSH的范围为 8.64± 1 .40 μmol/L ,GSSG的范围为 1 .0 8± 0 .2 4μmol/L ,GSH/GSSG的比值为1 0 .2 7± 1 .5 4;红细胞内GSH的范围为 6.81± 1 .3 8μmol/LRBC ,红细胞内GSSG的范围为 0 .77± 0 .3 9μmol/LR BC ,GSH/GSSG的比值为 9.0 2± 3 .61。血浆GSH/GSSG的氧化还原电位为Eh =-2 1 3 .9mV± 4.94mV ;RBC液的Eh =-2 2 1 .4mV± 3 .1 6mV ,血浆内和红细胞内的GSH/GSSG比值以及电位值都呈正态分布。血浆和红细胞在 -80℃条件下保存后对GSH和GSSG的测定有一定影响 (p <0 .0 5 )。结论 :本文建立的GSH/GSSG测定方法具有良好的重复性、准确性 ;

Objective: Measuring the oxidized and reduced glutathione in the plasma and red blood cells (RBC) and investigate the distribution of their redox potential in health young people. Methods The measuring method was modified from the way for determining oxidized and reduced glutathione in tissues and the oxidized and reduced glutathione in plasma and RBC of 42 health young subjects was determined by the improved method. The redox potentials of GSH/GSSG in plasma and in RBC were calculated. The modified method was also investigated for its efficiency.Results: The modified method was highly repetitive and had an optimistic recovery ratio(between 98.7% and 97.5%). The standard curve was strict linear. Measuring the GSH and GSSG in plasma and RBC in 42 healthy adults(mean aged 23.5)with this method, it was showed that the value of GSH in plasma was8.64±1.40μmol·L 1 , GSSG was 1.08±0.24μmol·L 1 ,the GSH/GSSG ratio was 10.27±1.54; the value of GSH in RBC was 6.81±1.38μmol·L 1 RBC, GSSG was 0.77±0.39μmol·L 1 RBC, GSH/GSSG was 9.02±3.61. The potentials of GSH/GSSG were: 213.9mV±4.94mV in plasma and 221.4mV±3.16mV in RBC. The distribution of the GSH/GSSG and their redox potential was normal distribution. The frozen store ( 80℃) of samples has some impacts on measuring results of GSH and GSSG in plasma and RBC(p<0.05). [WT5”HZ]Conclusions:[WT5”BZ] The method modified here was highly repetitive and accurate, the oxidized and reduced glutathione in plasma and RBC determined by this method showed similar results with other reports,the ratio of GSH/GSSG and their redox potential were normal distribution, which suggest the measuring GSH/GSSG and their redox potential could be a reliable items to measure the redox status of plasma.