title: [Effect of different concentrations of ammonium chloride on expression of hypoxia-inducible factor-1α in Chang liver cells].
abstract: Objective:To investigate the effect of treatment with different concentrations of ammonium chloride (NH4Cl) on the mRNA and protein expression of hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) in Chang liver cells.Methods:Normal Chang liver cells were cultured and treated continuously with different concentrations of NH4Cl (1.25, 2.5, and 5 mmol/L). The Chang liver cells cultured normally were used as controls. RNA and protein were extracted at the 5th, 10th, and 15th generations. Quantitative real-time PCR, immunohistochemistry, and Western blot were used to measure the expression of HIF-1α and VEGF.Results:Quantitative real-time PCR showed that compared with the control group, the NH4Cl treatment group showed significant increases in the mRNA expression of HIF-1α and VEGF across the 5th, 10th, and 15th generations (F= 90976.659/1300.218/1896.800 and 41825.754/2381.321/2591.954, allP< 0.05). The results of immunohistochemistry showed that compared with the control group, the NH4Cl treatment group showed significant increases in the protein expression of HIF-1α and VEGF across the 5th, 10th, and 15th generations (F= 376.709/1615.358/1350.120 and 904.789/5105.186/8644.498, allP< 0.05). As was shown by the results of Western blot, compared with the control group, the NH4Cl treatment group showed significant increases in the protein expression of HIF-1α and VEGF across the 5th, 10th, and 15th generations (F= 228.499/6051.974/183.219 and 5549.429/40187.665/120982.183, allP< 0.05).Conclusion:Ammonia can increase the expression of HIF-1α and its downstream gene VEGF in Chang liver cells and cause the phenomenon of pseudohypoxemia.
authors: Deparptment of Infection Disease, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
authors: H L
authors: HL
authors: Gu
authors: H Y
authors: HY
authors: Yang
authors: J J
authors: JJ
authors: Li
authors: Y
authors: Y
authors: Li
authors: X M
authors: XM
authors: Zhao
authors: Y J
authors: YJ
authors: Zhou
authors: X L
authors: XL
authors: Chen
authors: Q C
authors: QC
authors: Kan
authors: Z J
authors: ZJ
authors: Yu
mesh_terms: D015153:Blotting, Western; D002478:Cells, Cultured; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D008099:Liver / Q000166:cytology; D060888:Real-Time Polymerase Chain Reaction; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A; D000643:Ammonium Chloride
chemical_list: C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D012333:RNA, Messenger; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D000643:Ammonium Chloride