三氧化二砷对红细胞毒性及能量代谢的影响

Effects of Arsenic trioxide on toxicity and energy metabolism of RBCs

目的:研究三氧化二砷(arsenic trioxide,As_(2)O_(3))对红细胞(red blood cells,RBCs)毒性及能量代谢的影响。方法:用终浓度为0.2 mmol/L、0.4 mmol/L、0.8 mmol/L、1.6 mmol/L、3.2 mmol/L As_(2)O_(3)分别在37℃培养箱孵育红细胞48 h,利用酶标仪分别检测红细胞存活率及红细胞溶血率;用终浓度为2.5 mmol/L As_(2)O_(3)孵育红细胞5 h,利用液相色谱串联质谱法进行红细胞内靶向能量代谢产物相对定量分析。结果:0.2 mmol/L As_(2)O_(3)可提高红细胞存活率(P<0.05),0.4 mmol/L和0.8 mmol/L As_(2)O_(3)对红细胞存活率无明显影响(P>0.05),但高浓度(1.6 mmol/L和3.2 mmol/L)As_(2)O_(3)可降低红细胞的存活率(P<0.05);As_(2)O_(3)致红细胞IC 50为5202μmol/L;红细胞溶血率随As_(2)O_(3)浓度增高逐渐升高,但低浓度As_(2)O_(3)(0.2 mmol/L和0.4 mmol/L)对溶血率几乎没有影响(P>0.05),0.8 mmol/L组、1.6 mmol/L组和3.2 mmol/L组溶血率增高(P<0.05),但溶血率<6%,红细胞破坏不严重;能量代谢产物的检测结果表明,2.5 mmol/L As_(2)O_(3)37℃孵育红细胞5 h后,红细胞内的三磷酸腺苷(adenosine triphosphate,ATP)、丙酮酸(pyruvic acid,PA)、还原型烟酰胺腺嘌呤二核苷酸磷酸(reduced nicotinamide adenine dinucleotide phosphate,NADPH)含量均增高(P<0.05)。结论:低浓度As_(2)O_(3)可能通过促进红细胞糖酵解与磷酸戊糖途径保护红细胞。

Objective:To study the effects of Arsenic trioxide(As_(2)O_(3))on the toxicity and energy metabolism of RBCs.Methods:Firstly,RBCs were incubated with As_(2)O_(3)at the final concentration of 0.2 mmol/L,0.4 mmol/L,0.8 mmol/L,1.6 mmol/L and 3.2 mmol/L in 37℃incubator for 48 h,and then the cell viability and hemolysis rate of RBCs were measured by an enzyme labeling instrument.Finally,RBCs were incubated with As_(2)O_(3)at a final concentration of 2.5 mmol/L for 5 h,and the relative quantitative analysis of targeted energy metabolites in RBCs was performed by liquid chromatography-tandem mass spectrometry.Results:As_(2)O_(3)at final concentration of 0.2 mmol/L could improve the cell viability of RBCs,and the difference has statistically significant(P<0.05),0.4 mmol/L and 0.8 mmol/L As_(2)O_(3)had no significant effect on the viability of RBCs,but high concentration(1.6 mmol/L and 3.2 mmol/L)As_(2)O_(3)significantly reduced the viability of RBCs,and the difference has statistically significant(P<0.05).The half maximal inhibitory concentration(IC 50)of As_(2)O_(3)RBCs was 5202μmol/L.The hemolysis rate of RBCs increased gradually with the increasing concentration of As_(2)O_(3),but low As_(2)O_(3)concentration(0.2 mmol/L and 0.4 mmol/L)had little effect on the hemolysis rate,and the hemolysis rate at 0.8 mmol/L,1.6 mmol/L and 3.2 mmol/L groups were increased gradually(P<0.05),but the hemolysis rate was less than 6%,and destruction of seemed not very serious.The results of energy metabolites showed that the contents of adenosine triphosphate(ATP),pyruvate(PA),and nicotinamide adenine dinucleotide phosphate(NADPH)were increased significantly in RBCs after incubated with 2.5 mmol/L As_(2)O_(3)at 37℃for 5 h,and the difference had statistically significant(P<0.05).Conclusion:Low concentration of As_(2)O_(3)may protect RBCs by promoting glycolysis and pentose phosphate pathway.