I-Hui Yang1, Jhen-Hong Wong2, Che-Mai Chang3, Ben-Kuen Chen4, Yao-Ting Tsai2, Wei-Chiao Chen5, Eric Terry Wang6, Wen-Li Hsu7, Wei-Chiao Chang8. 1. Department of Ophthalmology, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan. 2. Department of Clinical Pharmacy, School of Pharmacy, Taipei Medical University, Taipei, Taiwan Department of Medical Genetics, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. 3. Department of Clinical Pharmacy, School of Pharmacy, Taipei Medical University, Taipei, Taiwan Master Program for Clinical Pharmacogenomics and Pharmacoproteomics, School of Pharmacy, Taipei Medical University, Taipei, Taiwan. 4. Institute of Bioinformatics and Biosignal Transduction, College of Bioscience and Biotechnology, National Cheng Kung University, Tainan, Taiwan. 5. Department of Clinical Pharmacy, School of Pharmacy, Taipei Medical University, Taipei, Taiwan. 6. Master Program for Clinical Pharmacogenomics and Pharmacoproteomics, School of Pharmacy, Taipei Medical University, Taipei, Taiwan. 7. Department of Medical Genetics, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. 8. Department of Clinical Pharmacy, School of Pharmacy, Taipei Medical University, Taipei, Taiwan Department of Pharmacy, Taipei Medical University Wan Fang Hospital, Taipei, Taiwan Comprehensive Cancer Center, Taipei Medical University, Taipei, Taiwan.
Abstract
PURPOSE: Calcium signaling is an important intracellular pathway. Increased intracellular calcium is associated with cytokine regulation and inflammatory signals secretion. The purpose of this study is to understand the molecular mechanisms by which calcium signaling controls IL-8 activation in human RPE cells. METHODS: Fluorescence-based calcium imaging and different mutants of IL-8 plasmids were used in this study. The IL-8 promoter activation, gene expression, and secretion were detected by using luciferase reporter assay, quantitative real-time PCR (Q-PCR), and ELISA, respectively. In addition, pharmacological inhibitors and small interfering RNA (siRNA) were applied to clarify the mechanisms of IL-8 activation. RESULTS: Our study reported that intracellular calcium mobilization activated IL-8 gene expression and secretion. Application of pharmacological inhibitor BAY 11-7082, siRNA, and plasmids of the nuclear factor κ light chain enhancer of activated B cells (NF-κB) binding site, we identified that NF-κB is the main transcription factor involved in intracellular calcium mobilization-mediated IL-8 activation in human RPE cells. CONCLUSIONS: Collectively, our findings highlight the important role of intracellular calcium mobilization in the activation of IL-8. These findings may be helpful for the clinical applications in the age-related macular degeneration (AMD) prevention and treatment. Copyright 2015 The Association for Research in Vision and Ophthalmology, Inc.
PURPOSE:Calcium signaling is an important intracellular pathway. Increased intracellular calcium is associated with cytokine regulation and inflammatory signals secretion. The purpose of this study is to understand the molecular mechanisms by which calcium signaling controls IL-8 activation in human RPE cells. METHODS: Fluorescence-based calcium imaging and different mutants of IL-8 plasmids were used in this study. The IL-8 promoter activation, gene expression, and secretion were detected by using luciferase reporter assay, quantitative real-time PCR (Q-PCR), and ELISA, respectively. In addition, pharmacological inhibitors and small interfering RNA (siRNA) were applied to clarify the mechanisms of IL-8 activation. RESULTS: Our study reported that intracellular calcium mobilization activated IL-8 gene expression and secretion. Application of pharmacological inhibitor BAY 11-7082, siRNA, and plasmids of the nuclear factor κ light chain enhancer of activated B cells (NF-κB) binding site, we identified that NF-κB is the main transcription factor involved in intracellular calcium mobilization-mediated IL-8 activation in human RPE cells. CONCLUSIONS: Collectively, our findings highlight the important role of intracellular calcium mobilization in the activation of IL-8. These findings may be helpful for the clinical applications in the age-related macular degeneration (AMD) prevention and treatment. Copyright 2015 The Association for Research in Vision and Ophthalmology, Inc.
Authors: Alana L Cutliffe; Sharon L McKenna; Darshan S Chandrashekar; Alvin Ng; Ginny Devonshire; Rebecca C Fitzgerald; Tracey R O'Donovan; John J Mackrill Journal: Explor Target Antitumor Ther Date: 2021-12-31