不同浓度葡萄糖对红细胞能量代谢的影响  

Effects of different concentrations of glucose on energy metabolism of red blood cells

作　　者：李旭妍 孙明玥 苏燕[1] 杨丽丽 王亚奇 宋芳[1] 闫巧梅[1] 黄世琪[1] 闫朝丽[2] 萨如拉[2] 李晶晶[3] 梁浩 王云霞 韩家鑫 聂欣雨 刘瑞刚[4] Li Xuyan;Sun Mingyue;Su Yan(Department of Biochemistry and Molecular Biology,Baotou Medical College,Baotou 014000)

机构地区：[1]包头医学院生物化学与分子生物学教研室,包头014000 [2]内蒙古医科大学第一附属医院内分泌科,呼和浩特010000 [3]包头医学院临床学院,包头014000 [4]包头医学院第一附属医院检验科,包头014000

出　　处：《安徽医科大学学报》2020年第11期1701-1706,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81860029);内蒙古自然科学基金(编号:2016MS0801);内蒙古自治区卫生和计划生育委员会科研计划项目(编号:201701090);内蒙古自治区研究生科研创新资助项目(编号:S2018112036Z);包头医学院“花蕾计划”项目(编号:2018BYJJ-HL-05、2018BYJJ-HL-25)。

摘　　要：目的探讨体外不同浓度葡萄糖对红细胞能量代谢的影响及其分子机制。方法采集6例健康成人新鲜抗凝血,离心去除白细胞和血浆,用PBS重悬洗涤红细胞3次后,将6组红细胞轻轻混匀后再分为4组,分别向其中加入D-葡萄糖溶液,使其终浓度分别为0、6、20、30 mmol/L,于37℃培养0、24、48 h后,取部分红细胞悬液利用流式细胞仪及多功能酶标仪检测红细胞内ATP含量、己糖激酶(HK)活性、丙酮酸激酶(PK)活性、活性氧(ROS)含量及红细胞衰亡率。实验结果采用单因素方差分析进行组间变量比较。结果与对应时间点的0 mmol/L组比较,用浓度为6、20、30 mmol/L的葡萄糖孵育红细胞24和48 h后,红细胞内HK活性、PK活性、ATP含量均增加,且随着葡萄糖浓度增加呈上升趋势,差异有统计学意义(P <0.05);而ROS含量和红细胞衰亡率是降低的,且随葡萄糖浓度增加而降低,差异有统计学意义(P <0.05)。用不同浓度葡萄糖(浓度分别为0、6、20、30mmol/L)培养红细胞48 h后,各不同浓度组红细胞内ROS含量和红细胞衰亡率随着孵育时间延长而增加,与对应浓度组的0 h比较,差异有统计学意义(P <0.05);HK活性和PK活性逐渐降低,与对应浓度组的0 h比较,差异有统计学意义(P <0.05)。而ATP含量呈波动式下降,与对应浓度组的0 h比较,差异有统计学意义(P <0.05)。结论红细胞体外短期高糖培养,不仅会加强红细胞内能量代谢,而且会降低氧化应激及衰亡发生率,而没有葡萄糖供应会导致红细胞内ATP不断被消耗,氧化应激增强,衰亡增多。Objective To investigate the effects of different concentrations of glucose on the energy metabolism of red blood cells(RBC)in vitro and its molecular mechanism.Method Fresh anticoagulant blood was collected from 6 healthy adults.The white blood cells and plasma were removed by centrifugation.After washing RBC three times with PBS respectively,the 6 people’s RBC were gently mixed and divided into 4 groups.D-glucose was added to make a final concentration of 0,6,20,30 mmol/L,respectively.After incubation at 37℃for 0,24 and 48 hours,some of the RBC suspension was taken out to detect eryptosis,ATP concentration,hexokinase(HK)activity,pyruvate kinase(PK)activity,reactive oxygen species(ROS)content and hemolysis rate by using flow cytometry and multi-function microplate reader.The experimental results were analyzed by one-way ANOVA.Results Compared with the 0 mmol/L group,after incubating RBC with glucose at 6,20,30 mmol/L for 24 h and 48 h,the HK activity,PK activity,and ATP content in the RBC all increased,and upward trend with increasing glucose concentration,and the difference was statistically significant(P<0.05);while ROS content and decay rate decreased,and decreased with increasing glucose concentration,the difference was statistically significant(P<0.05).After culturing erythrocytes with different concentrations of glucose(concentrations of 0,6,20,30 mmol/L)for 48 h,the ROS content and erythrocyte decay rate in RBC of different concentrations increased with the incubation time,compared with 0 h,the difference was statistically significant(P<0.05);HK activity and PK activity gradually decreased,compared with 0 h,the difference was statistically significant(P<0.05).The ATP content decreased in a fluctuating manner,and compared with 0 h,the difference was statistically significant(P<0.05).Conclusion Short-term high-glucose culture of RBC in vitro not only strengthens the energy metabolism in RBC,but also reduces the oxidative stress and eryptosis.However,no glucose supply leads to gradually exhausted ATP and

关 键 词：红细胞 葡萄糖 能量代谢 衰亡 

分 类 号：R589.9[医药卫生—内分泌]