• 생명과학회지
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• 제17권9호통권89호
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• pp.1204-1210
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• 2007

Totopoisomerase II 저해제인 etoposide는 핵안에 DNA double strand breaks를 일으키므로써 세포의 DNA에 손상을 초래한다. 본 연구에서는 인간 망막색소상피세포인 ARPE-19 세포에서의 세포성장 및 아폽토시스에서 etoposide의 역할을 살펴보았다. Etoposide는 세포의 성장을 크게 감소시켰으며 TUNEL에서 아폽토시스를 나타내는 DNA fragmentation의 증가를 유도하였다. 게다가, etoposide는 산화적 손상에 대해 세포나 조직을 보호하는 역할을 하는 것으로 알려진 세포내 항산화효소인 heme oxygenase-1 (HO-1)의 발현을 크게 증가시켰다. Etoposide에 의한 HO-1 발현증가는 항산화물질 NAC에 의해 억제되었는데, 이는 etoposide에 의한 세포내 ROS의 증가가 HO-1 발현에 중요한 역할을 한다는 것을 의미한다. 또한 HO-1 발현을 억제하기 위하여 HO-1 siRNA 방법을 사용하였다. 흥미롭게도, HO-1 유전자의 knock-down은 etoposide에 의해 유도되는 DNA fragmentation의 정도를 증가시켰다. 이들 결과를 종합해볼 때, etoposide에 의해 자극되어진 ARPE-19 세포에서 발현증가된 HO-1은 etoposide에 의한 아폽토시스 유발과정에서 세포를 보호하는 항아폽토시스의 기능을 한다는 것을 시사한다.

• Journal of Ginseng Research
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• 제47권1호
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• pp.65-73
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• 2023

Background: Age-related macular degeneration (AMD) is a significant visual disease that induces impaired vision and irreversible blindness in the elderly. However, the effects of ginseng berry extract (GBE) on the retina have not been studied. Therefore, this study aimed to investigate the protective effects of GBE on blue light (BL)-induced retinal damage and elucidate its underlying mechanisms in human retinal pigment epithelial cells (ARPE-19 cells) and Balb/c retina. Methods: To investigate the effects and underlying mechanisms of GBE on retinal damage in vitro, we performed cell viability assay, pre-and post-treatment of sample, reactive oxygen species (ROS) assay, quantitative real-time PCR (qRT-PCR), and western immunoblotting using A2E-laden ARPE-19 cells with BL exposure. In addition, Balb/c mice were irradiated with BL to induce retinal degeneration and orally administrated with GBE (50, 100, 200 mg/kg). Using the harvested retina, we performed histological analysis (thickness of retinal layers), qRT-PCR, and western immunoblotting to elucidate the effects and mechanisms of GBE against retinal damage in vivo. Results: GBE significantly inhibited BL-induced cell damage in ARPE-19 cells by activating the SIRT1/PGC-1α pathway, regulating NF-kB translocation, caspase 3 activation, PARP cleavage, expressions of apoptosis-related factors (BAX/BCL-2, LC3-II, and p62), and ROS production. Furthermore, GBE prevented BL-induced retinal degeneration by restoring the thickness of retinal layers and suppressed inflammation and apoptosis via regulation of NF-kB and SIRT1/PGC-1α pathway, cleavage of caspase 3 and PARP, and expressions of apoptosis-related factors in vivo. Conclusions: GBE could be a potential agent to prevent dry AMD and progression to wet AMD.

• Journal of Ginseng Research
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• 제47권2호
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• pp.311-318
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• 2023

Background: The beneficial effects of compound K (CK) on different chronic diseases have been shown to be at least related to antioxidant action. Nevertheless, since its antioxidant activity in human retinal pigment epithelial (RPE) cells is still unknown, here we investigated whether CK alleviates oxidative stress-stimulated damage in RPE ARPE-19 cells. Methods: The cytoprotective consequence of CK in hydrogen peroxide (H₂O₂)-treated cells was evaluated by cell viability, DNA damage, and apoptosis assays. Fluorescence analysis and immunoblotting were performed to investigate the inhibitory action of CK on reactive oxygen species (ROS) production and mitochondrial dysfunction. Results: H₂O₂-promoted cytotoxicity, oxidative stress, DNA damage, mitochondrial impairment, and apoptosis were significantly attenuated by CK in ARPE-19 cells. Furthermore, nuclear factor erythroid 2-related factor 2 (Nrf2) phosphorylation level and its shuttling to the nucleus were increased, which was correlated with upregulated activation of heme oxygenase-1 (HO-1). However, zinc protoporphyrin, a blocker of HO-1, significantly abrogated the preventive action of CK in H₂O₂-treated ARPE-19 cells. Conclusion: This study indicates that activation of Nrf2/HO-1 signaling by CK plays an important role in rescuing ARPE-19 cells from oxidative cellular damage.

• 생명과학회지
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• 제32권11호
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• pp.865-871
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• 2022

황반 변성은 현대 사회에서 발병율이 급격히 증가한 질환 중 하나이다. 황반 변성의 대표적인 원인은 노화로 인한 산화 스트레스로 널리 알려져 있고 최근 노령화로 인해 발병 연령은 40대부터 급격하게 증가하고 있다. 따라서 본 연구에서는 TMH를 이용해 산화적 스트레스에 대한 항산화 효능 및 세포 사멸 억제연구를 통해 눈 건강 개선 소재를 개발하였다. 먼저 TMH의 인간 망막색소상피세포 사멸 억제 효능 평가를 위해 MTS assay로 세포 생존율을 측정하였다. 망막색소상피세포에서 H₂O₂에 의한 산화적 스트레스로 인해 약 60%의 세포 생존율을 확인 할 수 있었고 TMH에 의해 농도 의존적으로 세포 사멸을 억제됨을 확인할 수 있었다. 또한 LDH release assay를 통해 세포막 보호 효능을 확인 하였으며 세포 내 ROS 측정을 통해 세포 내 활성산소종 조절을 통한 항산화 효능을 확인 할 수 있었다. 마지막으로 세포 생존과 사멸에 관여하는 세포 내 신호 단백질인 MAPKs의 인산화 조절, 항산화 효소인 HO-1 발현 조절, 세포 사멸 관여 단백질인 Bax/Bcl-2의 발현 그리고 cleaved caspase-3 단백질 분석을 통해 TMH의 산화 스트레스에 대한 항산화 그리고 세포 사멸 억제 경로를 확인 하였다. 위와 같은 결과 도출을 통해 TMH의 눈 건강 기능성 소재로서의 가능성을 확인하였다.

• Parasites, Hosts and Diseases
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• 제57권2호
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• pp.83-92
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• 2019

Based on the reactive oxygen species (ROS) regulatory properties of diphenyleneiodonium (DPI), we investigated the effects of DPI on host-infected T. gondii proliferation and determined specific concentration that inhibit the intracellular parasite growth but without severe toxic effect on human retinal pigment epithelial (ARPE-19) cells. As a result, it is observed that host superoxide, mitochondria superoxide and $H_2O_2$ levels can be increased by DPI, significantly, followed by suppression of T. gondii infection and proliferation. The involvement of ROS in anti-parasitic effect of DPI was confirmed by finding that DPI effect on T. gondii can be reversed by ROS scavengers, N-acetyl-L-cysteine and ascorbic acid. These results suggest that, in ARPE-19 cell, DPI can enhance host ROS generation to prevent T. gondii growth. Our study showed DPI is capable of suppressing T. gondii growth in host cells while minimizing the un-favorite side-effect to host cell. These results imply that DPI as a promising candidate material for novel drug development that can ameliorate toxoplasmosis based on ROS regulation.

• 한국축산식품학회지
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• 제44권3호
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• pp.699-709
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• 2024

Oxya chinensis sinuosa (OC) is a well-known edible insect. Several researches on the health benefits of OC consumption have been performed to date; however, their effect on eye health remains largely unknown. This study aimed to assess the protective effects of OC extracts on the oxidative stress on the retinal pigment epithelium (RPE) cells. Oxidative damage has been identified as one of the key regulatory factors in agerelated macular degeneration. H₂O₂-induced reactive oxygen species (ROS) production, a well-known oxidative stress factor, can cause cell death in retinal pigment epithelia cells. In this study, we found that three OC extracts effectively prevented H₂O₂-induced ROS production and subsequent death of ARPE-19 cells in a dose-dependent manner. In addition, the OC extracts inhibited the phosphorylation of mitogen-activated protein kinases including p38, JNK, and ERK. The OC extracts restored IκBα degradation induced by H₂O₂, indicating that OC extracts suppressed the activation of nuclear factorκB. Furthermore, the three OC extracts were shown to have antioxidant effects by upregulating the intracellular expression of key antioxidant proteins such as SOD, NQO, and HO-1. Here we demonstrated the antioxidant and anti-apoptotic effects of the OC extracts on ARPE-19, indicating their potential role in improving eye health. These results suggest that three OC extracts plays a critical role in oxidative stress-induced cell death protects in ARPE-19 cells.

• International Journal of Industrial Entomology and Biomaterials
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• 제47권2호
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• pp.90-98
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• 2023

The human eye, constantly exposed to solar radiation, can be damaged by UV radiation. In particular, ultraviolet B (UVB)-induced damage plays an important role in retinal degeneration and cell aging. In this study, we investigated the protective effects of the methanol extract of Oxya chinensis sinuosa (OCM), an edible insect known for its high protein content (64.2%), and various pharmacological effects, on human retinal pigment epithelial cells. ARPE-19 cells were treated with OCM and subsequently UVB irradiated. Our results showed that OCM effectively attenuates UVB-induced cell damage by reducing MAPK phosphorylation (JNK and p38 MAPK). Additionally, OCM increased the phosphorylation of Akt, and cell cycle regulators, including p21 and p27, in a dose-dependent manner. Moreover, OCM treatment increased ARPE-19 cell proliferation by activating the S6K1/S6 pathway. This study suggests that OCM prevents UVB-induced retinal cell damage by increasing cell proliferation via ROS reduction, suggesting its potential as a functional therapeutic superfood against retinal cell damage.

• The Korean Journal of Physiology and Pharmacology
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• 제27권3호
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• pp.231-240
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• 2023

Fabry disease is a lysosomal storage disorder characterized by the lysosomal accumulations of glycosphingolipids in a variety of cytotypes, which include endothelial cells. The disease is inherited and originates from an error in glycosphingolipid catabolism caused by insufficient α-galactosidase A activity, which causes uncontrolled progressive storage of intracellular globotriaosylceramide (Gb3) in the vasculature and extracellular accumulation of lyso-Gb3 (a deacetylated soluble form of Gb3). Necrosis can lead to inflammation, which exacerbates necrosis and creates a positive feedback loop that triggers necroinflammation. However, the role played by necroptosis, a form of programmed necrotic cell death, in the cell-to-cell inflammatory reaction between epithelial and endothelial cells is unclear. Thus, the present study was undertaken to determine whether lyso-Gb3 induces necroptosis and whether necroptosis inhibition protects endothelial dysfunction against lyso-Gb3 inflamed retinal pigment epithelial cells. We found lyso-Gb3 induced necroptosis of a retinal pigment epithelial cell line (ARPE-19) in an autophagy-dependent manner and that conditioned media (CM) from ARPE-19 cells treated with lyso-Gb3 induced the necroptosis, inflammation, and senescence of human umbilical vein endothelial cells. In addition, a pharmacological study showed CM from lyso-Gb3 treated ARPE-19 cells induced endothelial necroptosis, inflammation, and senescence were significantly inhibited by an autophagy inhibitor (3-MA) and by two necroptosis inhibitors (necrostatin and GSK-872), respectively. These results demonstrate lyso-Gb3 induces necroptosis via autophagy and suggest that lyso-Gb3 inflamed retinal pigment epithelial cells trigger endothelial dysfunction via the autophagy-dependent necroptosis pathway. This study suggests the involvement of a novel autophagy-dependent necroptosis pathway in the regulation of endothelial dysfunction in Fabry disease.

• 생명과학회지
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• 제27권11호
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• pp.1349-1356
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• 2017

$K^+$ 통로 개방제들은 심근, 뇌, 골격근 등에서 세포막 혹은 미토콘드리아 내막에 존재하는 큰 전도성의 $Ca^{2+}$-의 존성 $K^+$ (BK) 통로 및 ATP-조절성 $K^+$ 통로(ATP-sensitive $K^+$ channels, $K_{ATP}$)에 작용하여 허혈성 혹은 산화성 세포 손상을 완화하는 효과가 있는 것으로 보고되어 있다. 본 연구에서는 망막 색소 상피세포주인 ARPE-19 세포를 실험 모델로 하여 큰 전도성의 BK 통로 개방제인 NS 1619가 유사한 보호 효과를 나타낼 수 있는지, 또한 그 작용기전이 무엇인지를 확인하고자 하였다. AREE-19 세포를 여러 형태의 산화 스트레스에 노출시켜 세포 손상을 유발하고 그 손상의 정도 및 이에 미치는 NS 1619의 효과를 trypan blue 배출능, Tunel 염색 분석을 통하여 측정하였다. NS 1619는 여러 형태의 산화 스트레스에 의한 괴사성 및 apoptosis에 의한 세포 손상을 효과적으로 방지하였으며 그 보호 효과는 BK 통로 봉쇄제인 paxilline 의해 차단되었다. NS 1619는 $H_2O_2$에 의한 세포내 ATP 고갈을 현저히 완화시켰으며, 또한 MTT 환원능으로 측정한 미토콘드리아의 기능을 보호하는 효과를 보였다. 유세포형광 분석법을 이용한 실험에서 NS 1619는 $H_2O_2$에 의한 미토콘드리아 막전압의 소실을 유의하게 방지하였다. 이상의 결과들을 종합하면 NS 1619는 망막 색소 상피세포에서 산화성 세포 손상을 방지하는 효과를 나타내며 그 기전에 미토콘드리아 기능에 대한 보호 작용이 연관되어 있는 것으로 사료된다.

• KIEE International Transactions on Electrophysics and Applications
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• 제4C권4호
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• pp.160-164
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• 2004

In this study, we demonstrate for the first time a bio-inspired Cell-Microsystem to manipulate and detect living cells. Cultured retinal pigment epithelial cell line (ARPE-19) was directed to grow in a pre-defined Cell-Microsystem. The three-dimensional micropillars of 5 ${\mu}{\textrm}{m}$ in height and diameter of the Cell-Microsystem were fabricated. Inhibited DNA synthesis and transformed cell morphology were observed throughout the culture period. The demonstration of manipulating and detecting living cells by the surface topography is a new approach, and it will be very useful for the future design of cell-based biosensors and bioactuators.