Objective: To explore the pathogenesis of erythrocytosis by detecting the key enzymes of glucose metabolism and glucose transporter in bone marrow erythrocytes of chronic mountain sickness (CMS), and analyzing its correlation with hemoglobin.
Methods: Twenty CMS patients hospitalized in Qinghai Provincial People's Hospital from January 2019 to December 2020 were selected as CMS group. Twenty males with leukocyte count > 3.5×109/L who had accepted bone marrow aspiration and had normal result were taken as control group. The mRNA and protein expression of key enzymes and glucose transporter in glucose metabolism in bone marrow CD71+ erythrocytes were detected by real time qPCR and Western blot, respectively. Glucose, lactic acid and 2,3-diphosphoglycerate in the bone marrow supernatant and serum were tested by ELISA. The mRNA and protein expression of key enzymes and glucose transporter, glucose, lactic acid and 2,3-diphosphoglycerate of the two groups were compared. Pearson correlation was used to analyze the correlation between key enzymes, glucose transporter in glucose metabolism in bone marrow CD71+ erythrocytes and hemoglobin.
Results: The expression of HK2, GLUT1 and GLUT2 mRNA in the CMS group were higher than those in the control group (P<0.001), while the expression of HK1, OGDH and COX5B mRNA were not different. The expression of HK2, GLUT1 and GLUT2 protein in the CMS group were higher than those in the control group (P<0.05). The levels of glucose and lactic acid in the bone marrow supernatant and serum in the CMS group were not different from those in the control group, while the level of 2,3-diphosphoglycerate was higher (P<0.001). Both HK2 and GLUT2 proteins were positively correlated with hemoglobin (r=0.511, 0.717).
Conclusion: CMS patients may increase glycolysis by increasing the expression of HK2, and promote the utilization of glucose through high expression of GLUT1 and GLUT2 to meet the need of energy supply.
题目: 慢性高原病糖代谢关键酶的表达以及与表型间的关系.
目的: 通过检测慢性高原病（CMS）骨髓红细胞中糖代谢的关键酶和葡萄糖转运蛋白，分析其与血红蛋白的关系，探求红细胞增多的发病机制.
方法: 选取青海省人民医院2019年1月至2020年12月住院的20例CMS患者作为CMS组，白细胞>3.5×109/L的20例愿意接受骨髓穿刺且结果正常的男性作为对照组，PCR检测骨髓CD71+细胞中糖代谢关键酶和葡萄糖转运蛋白mRNA的表达，蛋白免疫印迹法检测骨髓CD71+细胞中糖代谢关键酶和葡萄糖转运蛋白的表达，ELISA检测骨髓上清及血清中葡萄糖、乳酸及2,3-二磷酸甘油酸水平，比较两组糖代谢关键酶和葡萄糖转运蛋白mRNA和蛋白的水平，以及葡萄糖、乳酸及2,3-二磷酸甘油酸水平，采用Pearson相关分析糖代谢关键酶和葡萄糖转运蛋白与血红蛋白之间的相关性.
结果: CMS组HK2、GLUT1和GLUT2 mRNA的表达水平高于对照组（P<0.001），而HK1、OGDH和COX5B mRNA与对照组比较无差异；CMS组HK2、GLUT1和GLUT2蛋白的表达水平均高于对照组（P<0.05）；CMS组骨髓上清及血清葡萄糖、乳酸的水平与对照组比较无差异，而骨髓上清2,3-二 磷酸甘油酸水平高于对照组（P<0.001）。HK2和GLUT2蛋白与血红蛋白之间均呈正相关（r=0.511，0.717）.
结论: CMS患者可通过增加HK2的表达增加糖酵解，通过GLUT1和GLUT2的高表达促进葡萄糖的利用以满足供能的需要.
Keywords: chronic mountain sickness; enzyme; glucose metabolism; phenotype.