目的: 研究三氧化二砷(arsenic trioxide,As₂O₃)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 As₂O₃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 As₂O₃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 As₂O₃可提高红细胞存活率(P＜0.05),0.4 mmol/L和0.8 mmol/L As₂O₃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)As₂O₃可降低红细胞的存活率(P＜0.05);As₂O₃致红细胞IC₅₀为5 202 μmol/L;红细胞溶血率随As₂O₃浓度增高逐渐升高,但低浓度As₂O₃(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 As₂O₃ 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As₂O₃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 (As₂O₃) on the toxicity and energy metabolism of RBCs. Methods: Firstly, RBCs were incubated with As₂O₃ 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 As₂O₃ 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: As₂O₃ 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 As₂O₃ had no significant effect on the viability of RBCs, but high concentration (1.6 mmol/L and 3.2 mmol/L) As₂O₃ significantly reduced the viability of RBCs, and the difference has statistically significant(P＜0.05). The half maximal inhibitory concentration (IC₅₀) of As₂O₃ RBCs was 5 202 μmol/L. The hemolysis rate of RBCs increased gradually with the increasing concentration of As₂O₃, but low As₂O₃ 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 As₂O₃ at 37℃ for 5 h, and the difference had statistically significant (P＜0.05). Conclusion: Low concentration of As₂O₃ may protect RBCs by promoting glycolysis and pentose phosphate pathway.