3,3'-Diindolylmethane inhibits VEGF expression through the HIF-1α and NF-κB pathways in human retinal pigment epithelial cells under chemical hypoxic conditions

Park,Hongzoo; Lee,Dae-Sung; Yim,Mi-Jin; Choi,Yung  Hyun; Park,Saegwang; Seo,Su-Kil; Choi,Jung  Sik; Jang,Won  Hee; Yea,Sung  Su; Park,Won  Sun; Lee,Chang-Min; Jung,Won-Kyo; Choi,Il-Whan

doi:10.3892/ijmm.2015.2202

3,3'-Diindolylmethane inhibits VEGF expression through the HIF-1α and NF-κB pathways in human retinal pigment epithelial cells under chemical hypoxic conditions

Authors:
- Hongzoo Park
- Dae-Sung Lee
- Mi-Jin Yim
- Yung Hyun Choi
- Saegwang Park
- Su-Kil Seo
- Jung Sik Choi
- Won Hee Jang
- Sung Su Yea
- Won Sun Park
- Chang-Min Lee
- Won-Kyo Jung
- Il-Whan Choi
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Affiliations: Department of Urology, Kangwon National University Hospital, Kangwon National University School of Medicine, Chuncheon, Gangwon-do, Republic of Korea, Marine Biodiversity Institute of Korea, Seocheon, Chungcheongnam-do, Republic of Korea, Department of Biochemistry, College of Oriental Medicine, Dongeui University, Busan, Republic of Korea, Department of Microbiology, College of Medicine, Inje University, Busan, Republic of Korea, Department of Internal Medicine, Busan Paik Hospital, College of Medicine, Inje University, Busan, Republic of Korea, Department of Biochemistry, College of Medicine, Inje University, Busan, Republic of Korea, Department of Physiology, Kangwon National University School of Medicine, Chuncheon, Gangwon-do, Republic of Korea, Department of Molecular Microbiology and Immunology, Warren Alpert School of Medicine, Providence, RI, USA, Department of Biomedical Engineering and Center for Marine-Integrated Biomedical Technology (BK21 Plus), Pukyong National University, Busan, Republic of Korea
Published online on: May 5, 2015 https://doi.org/10.3892/ijmm.2015.2202
Pages: 301-308

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Abstract

Oxidative stress in the retinal pigment epithelium (RPE) can lead to the pathological causes of age-related macular degeneration (AMD). Hypoxia induces oxidative damage in retinal pigment epithelial cells (RPE cells). In this study, we investigated the capacity of 3,3'-diindolylmethane (DIM) to reduce the expression of vascular endothelial growth factor (VEGF) under hypoxic conditions, as well as the molecular mechanisms involved. Human RPE cells (ARPE-19 cells) were treated with cobalt chloride (CoCl2, 200 µM) and/or DIM (10 and 20 µM). The production of VEGF was measured by enzyme-linked immunosorbent assay. The translocation of hypoxia-inducible factor-1α (HIF-1α) and nuclear factor-κB (NF-κB) was determined by western blot analysis. The binding activity of HIF-1α and NF-κB was analyzed by electrophoretic mobility shift assay. The phosphorylation levels of mitogen-activated protein kinases (MAPKs) were measured by western blot analysis. The levels of mitochondrial reactive oxygen species (ROS) were detected by fluorescence microplate assay. The results revealed that DIM significantly attenuated the CoCl2-induced expression of VEGF in the ARPE-19 cells. The CoCl2-induced translocation and activation of HIF-1α and NF-κB were also attenuated by treatment with DIM. In addition, DIM inhibited the CoCl2-induced activation of p38 MAPK in the ARPE-19 cells. Pre-treatment with YCG063, a mitochondrial ROS inhibitor, led to the downregulation of the CoCl2-induced production of VEGF by suppressing HIF-1α and NF-κB activity. Taken together, the findings of our study demonstrate that DIM inhibits the CoCl2-induced production of VEGF by suppressing mitochondrial ROS production, thus attenuating the activation of HIF-1α and p38 MAPK/NF-κB.

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