不同浓度谷胱甘肽对储存红细胞内氧化应激反应的影响

Effect of different concentrations of glutathione on oxidative stress response in stored red blood cells

目的探讨不同浓度谷胱甘肽(GSH)对储存红细胞(sRBC)内氧化应激反应的影响。方法选择6份新鲜制备的sRBC各取12 ml,平均分为四组:枸橼酸葡萄糖(ACD)组、GSH1组、GSH2组和GSH3组,每组3 ml。ACD组中加入ACD-B保存液1 ml,再加入生理盐水50μl,GSH1、GSH2和GSH3三组均加入ACD-B保存液1 ml,再分别加入12.5、62.5、125μmol/L的GSH溶液50μl。四组均置于4℃冰箱密封保存。在保存的第1、7、14、21天,分别采用酶联免疫吸附试验(ELISA)测定sRBC内ATP含量和GSH-PX酶活性,采用黄嘌呤氧化酶法测定sRBC内超氧化物歧化酶(SOD)活性,采用硫代巴比妥酸比色法测定sRBC内丙二醛(MDA)含量。结果与ACD组比较,第1、7、14、21天GSH2组和GSH3组的ATP含量和GSH-PX酶活性明显升高(P<0.05);与GSH1组比较,GSH2组和GSH3组ATP含量和GSH-PX酶活性明显升高(P<0.05)。与ACD组比较,第1、7、14、21天GSH2组SOD酶活性明显升高(P<0.05),第1、7、14天GSH3组SOD酶活性明显升高(P<0.05);与GSH1组比较,第21天GSH2组和GSH3组SOD酶活性明显升高(P<0.05)。与ACD组比较,第1、7、14、21天GSH2和GSH3组MDA含量明显降低(P<0.05);与GSH1组比较,第1、7、21天GSH2组和GSH3组MDA含量明显降低(P<0.05)。结论sRBC内加入62.5、125μmol/L的谷胱甘肽具有一定的抗氧化应激作用,可降低sRBC的储存性损伤。

Objective To observe the effect of different concentrations of glutathione(GSH)on oxidative stress response in stored red blood cells(sRBC).Methods sRBC 12 ml were taken from each of the six freshly prepared sRBC,and were divided into four groups:group ACD,group GSH1,group GSH2 and group GSH3.The acid-citrate-dextrose-B fluid(ACD-B)1 ml and normal saline 50μl were added in all the groups.GSH 50μl were added into the sRBC in groups GSH1,GSH2,and GSH3,and the final concentrations of GSH were 12.5,62.5 and 125μmol/L respectively.All of the sRBC of the four groups were stored in the refrigerator at 4℃.On the 1st,7th,14th,and 21st day of preservation,the contents of ATP and the activity of GSH-PX in the four groups of sRBC were determined by ELISA.The activity of SOD in sRBC in the four groups were determined using the xanthine oxidase method.The contents of MDA in sRBC in the four groups were determined using thiobarbituric acid colorimetric method.Results Compared with the group ACD,the content of ATP and the activity of GSH-PX in groups GSH2 and GSH3 were significantly increased on the 1st,7th,14th,and 21st day of preservation(P<0.05).Compared with the group GSH1,the content of ATP and the activity of GSH-PX in groups GSH2 and GSH3 were significantly increased on the 1st,7th,14th,and 21st day of preservation(P<0.05).Compared with group ACD,the activity of SOD in group GSH2 was significantly increased on the 1st,7th,14th,and 21st day(P<0.05),the activity of SOD in group GSH3 was significantly increased on the 1st,7th,and 14th day of preservation(P<0.05).Compared with group GSH1,the activity of SOD in groups GSH2 and GSH3 significantly increased on the 21st day of preservation(P<0.05).Compared with group ACD,the content of MDA in the groups GSH2 and GSH3 significantly decreased on the 1st,7th,14th,and 21st day of preservation(P<0.05).Compared with group GSH1,the content of MDA in the groups GSH2 and GSH3 significantly decreased on the 1st,7th,and 21st day(P<0.05).Conclusion The addition of GSH of 62.5 and 125μmol/L can improve oxidative stress response,and reduce the storage damage of sRBC.