Human retinal pigment epithelial cell proliferation by the combined stimulation of hydroquinone and advanced glycation end-products via up-regulation of VEGF gene

Abstract

Although recent research showed that advanced glycation endproduct (AGE) and hydroquinone (HQ) are related to the pathogenesis of age-related macular degeneration (AMD), the mechanism how AGE and HQ induce or accelerate AMD remains elusive. In the present study, we examined the effects of AGE and HQ on changes of human retinal pigment epithelial (RPE) cell numbers and found that the viable cell numbers were markedly reduced by HQ by apoptosis and that AGE prevented the decreases of HQ-treated cell numbers by increased replicative DNA synthesis of RPE cells without changing apoptosis. Real-time RT-PCR revealed that vascular endothelial growth factor (VEGF)-A mRNA was increased by HQ treatment and the addition of HQ+AGE resulted in a further increment. The increase of VEGF secretion was confirmed by ELISA, and inhibition of VEGF signaling by chemical inhibitors and small interfering RNA decreased the HQ+AGE-induced increases in RPE cell numbers. The deletion analysis demonstrated that −102 to −43 region was essential for the VEGF-A promoter activation. Site-directed mutaions of specificity protein 1 (SP1) binding sequences in the VEGF-A promoter and RNA interference of SP1 revealed that SP1 is an essential transcription factor for VEGF-A expression. These results indicate that HQ induces RPE cell apoptosis, leading to dry AMD, and suggest that AGE stimulation in addition to HQ enhances VEGF-A transcription via the AGE-receptor for AGE pathway in HQ-damaged cells. As a result, the secreted VEGF acts as an autocrine/paracrine growth factor for RPE and/or adjacent vascular cells, causing wet AMD.

Resvega Alleviates Hydroquinone-Induced Oxidative Stress in ARPE-19 Cells

“…Cigarette smoke destroys the macular area by increasing oxidative stress and suppressing the antioxidative system [9]. Hydroquinone is one component in the cigarette smoke, which is known to induce oxidative stress and apoptosis, and compromise cell viability concentration-dependently in RPE cells [16,21,27,43]. Our results are in line with the observation that hydroquinone-induced cytotoxicity is concentration-dependent and increases oxidative stress in RPE cells [16][17][18].…”

Effect of resveratrol on cancer progression through the REG III expression pathway in head and neck cancer cells

“…Total RNA was isolated using RNeasy Protect ® Cell Mini kit (Qiagen, Hiden, Germany) from FaDu cells. cDNA was reverse transcribed from 0.5-2 µg samples of total RNA using High-Capacity cDNA Reverse Transcription kit (Applied biosystems, Foster City, CA, uSA) as described (44,45,(47)(48)(49)(50)(51). cDNA was subjected to PCR with the following primers, synthesized and prepared by Nihon Gene Research Laboratories, Inc. (NGRL; Sendai, japan): β-actin (NM_001101) sense, 5'-GCGAGAAGATGACCCAGA-3' and antisense, 5'-CAGAGGCGTACAGGGATA-3'; REG Ⅲ (Ab161037) sense 5'-GAATATTCTCCCCAAACTG-3' and antisense, 5'-GAGAAAAGCCTGAAATGAAG-3'.…”

Statins decrease vascular epithelial growth factor expression via down-regulation of receptor for advanced glycation end-products

“…However, the molecular mechanism by which statins reduce the risk of AMD remains unclear. Recently, we reported that the combination of hydroquinone (HQ; benzene-1,4-diol) and advanced glycation endproducts (AGE) increased the expression of VEGF-A in RPE cells and that VEGF induced RPE cell proliferation as an autocrine growth factor [8] . In this study, we examined the effects of statins on the expression of VEGF-A and on receptor for AGE ( RAGE ) using the HQ + AGE-induced in vitro AMD model.…”