Development of a novel mouse model of severe glucose-6-phosphate dehydrogenase (G6PD)-deficiency for in vitro and in vivo assessment of hemolytic toxicity to red blood cells

详细信息    查看全文

• 作者：Chun Hay  ; Ko^a ;   ; ^b ; Karen  ; Li^b ; Chung Leung  ; Li^c ; Pak Cheung  ; Ng^b ; Kwok Pui  ; Fung^a ; Anthony Edward  ; James^d ; Raymond Pui-On  ; Wong^b ; Goldie Jia-Shi  ; Gu^b ; Tai Fai  ; Fok^a ;   ; ^{taifaifok@cuhk.edu.hk}
• 关键词：G6PD-deficiency ; Mouse ; Oxidative stress ; Glutathione ; Methemoglobin
• 刊名：Blood Cells, Molecules, and Diseases
• 出版年：2011
• 出版时间：15 October 2011
• 年：2011
• 卷：47
• 期：3
• 页码：176-181
• 全文大小：510 K

文摘

Studies of hemolytic agents on G6PD-deficient subjects have been extensively performed on red blood cells obtained from donors, only using in vitro methods. However, there has been no adequate G6PD-deficient animal model for in vivo assessment of potentially hemolytic agents. The objective of this study is to establish a novel mouse model of severe G6PD-deficiency, with high susceptibility to hemolytic damage upon oxidative agents. To create this model, G6PD mutant Gpdx allele was introduced into the C57L/J mouse strain background by breeding program. The hemolytic toxicity of naphthalene and its metabolite ¦Á-naphthol on G6PD-deficient red blood cells was evaluated. Our data showed that the F2 homozygous Gpdx mutant with C57L/J background exhibiting the G6PD activity was 0.9 ¡À 0.1 U/g Hb, level similar to those of G6PD deficiency in human. A significantly negative correlation was demonstrated between GSH percentage reduction and G6PD activity (r = ?0.51, p < 0.001) upon challenge of the red blood cells with alpha-naphthol in vitro. Similar correlation was also found between GSSG elevation and G6PD activity. Our in vivo studies showed that the administration of naphthalene at 250 mg/kg inflicted significant oxidative damage to the G6PD-deficient mice, as illustrated by the decrease of the GSH-to-GSSG ratio (by 34.2 % , p = 0.005) and the increase of the methemoglobin level (by 1.9 fold, p < 0.001). Hemolytic anemia was also found in G6PD-deficient mice at this dosage of naphthalene. In summary, this novel mouse model could be utilized as a screening platform to more accurately determine the hemolytic toxicity of pharmacological agents on G6PD-deficient subjects.