目的: 探讨砷暴露引起肺组织纤维化过程中是否发生有氧糖酵解的改变。方法: 在动物水平上,通过小鼠自由饮用含50 mg/L NaAsO2的纯净水,于染毒9个月后处死,制备肺组织切片镜下观察病理改变;细胞水平上,将HELF细胞暴露于不同浓度的NaAsO2(0、10、20、40 μmol/L)48 h,检测细胞活力、葡萄糖消耗量,使用qPCR和Western blot检测己糖激酶2(hexokinase 2, HK2)、乳酸脱氢酶A(lactate dehydrogenase A, LDHA)基因表达情况。结果: 小鼠经过9个月的砷暴露后,对照组肺泡结构完整,暴露组出现气道周围组织增厚、胶原沉积、肺泡壁增厚现象;小鼠NaAsO2暴露9个月后肺组织中有氧糖酵解关键酶HK2、LDHA的mRNA以及蛋白相对表达量与对照组相比升高(P＜0.05)。在细胞水平上,暴露48 h后,相较于对照组,NaAsO2浓度为10、20、40 μmol/L时HELF细胞活力降低(P＜0.05),葡萄糖消耗量增加(P＜0.05);在RNA水平上,随着暴露剂量增加HK2、LDHA表达升高(P＜0.05)。在蛋白水平上,随着暴露剂量增加HK2、LDHA表达升高(P＜0.05)。结论: 有氧糖酵解可能参与砷暴露引起肺成纤维细胞活化的过程。
Objective: To investigate whether changes in aerobic glycolysis occur during the process of lung tissue fibrosis induced by arsenic exposure. Methods: At the animal level, mice were allowed to freely drink pure water containing 50 mg/L NaAsO2. After nine months, the mice were sacrificed, and lung tissue sections were prepared for microscopic observation of changes. At the cellular level, HELF cells were exposed to different concentrations of NaAsO2 (0, 10, 20, 40 μmol/L) for 48 hours. Cell viability, glucose consumption were detected, and the gene expression of HK2 and LDHA was detected using qPCR and Western blot. Results: After nine months of arsenic exposure in mice, the alveolar structure of the control group was intact, while the exposed group showed thickening of the tissues around the airways, collagen deposition, and thickening of the alveolar walls. After nine months of NaAsO2 exposure, the relative expression of mRNA and protein of key enzymes of aerobic glycolysis HK2 and LDHA in lung tissue of mice increased compared with the control group (P＜0.05). At the cellular level, after 48 h of exposure, compared with the control group, the viability of HELF cells decreased when the concentration of NaAsO2 was 10, 20, 40 μmol/L (P＜0.05). Glucose consumption increased (P＜0.05). At the RNA level, the expression of HK2 and LDHA increased with the increase of exposure dose (P＜0.05). At the protein level, the expression of HK2 and LDHA increased with the increase of exposure dose (P＜0.05). Conclusion: Aerobic glycolysis may be involved in the process of activation of lung fibroblasts induced by arsenic exposure.
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