• 폴리머
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• 제38권3호
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• pp.364-370
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• 2014

망막색소상피(RPE)는 건강한 망막을 유지하는데 중요한 역할을 하고 RPE의 퇴화는 많은 망막질병을 유발한다. RPE 이식은 최근 망막 퇴화에 대한 가능성 있는 치료법으로 제시되고 있다. RPE 세포를 안전하게 이식하기 위해서는 지지체가 필요하므로 독특한 기계적 성질과 생체적합성을 갖는 실크를 사용하여 필름을 제조하였다. 실크필름의 FTIR, 접촉각 및 생분해성을 측정한 후, RPE 세포를 실크필름에 파종하여 그 영향을 확인하였다. MTT 분석, SEM, 면역형광염색, RT-PCR을 통해 세포의 부착, 생존도, 형태유지, 특이적 mRNA의 발현을 분석하였다. 본 연구에서는, 실크필름에 배양한 RPE 세포의 부착, 증식 및 표현형 유지가 뛰어남을 확인함으로써 실크필름의 망막 재생을 위한 조직 공학적 지지체로의 응용 가능성을 제시했다.

• Molecules and Cells
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• 제43권7호
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• pp.632-644
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• 2020

The molecular mechanism underlying autophagy impairment in the retinal pigment epithelium (RPE) in dry age-related macular degeneration (AMD) is not yet clear. Based on the causative role of poly(ADP-ribose) polymerase 1 (PARP1) in RPE necrosis, this study examined whether PARP1 is involved in the autophagy impairment observed during dry AMD pathogenesis. We found that autophagy was downregulated following H₂O₂-induced PARP1 activation in ARPE-19 cells and olaparib, PARP1 inhibitor, preserved the autophagy process upon H₂O₂ exposure in ARPE-19 cells. These findings imply that PARP1 participates in the autophagy impairment upon oxidative stress in ARPE-19 cells. Furthermore, PARP1 inhibited autolysosome formation but did not affect autophagosome formation in H₂O₂-exposed ARPE-19 cells, demonstrating that PARP1 is responsible for impairment of late-stage autophagy in particular. Because PARP1 consumes NAD⁺ while exerting its catalytic activity, we investigated whether PARP1 impedes autophagy mediated by sirtuin1 (SIRT1), which uses NAD⁺ as its cofactor. A NAD⁺ precursor restored autophagy and protected mitochondria in ARPE-19 cells by preserving SIRT1 activity upon H₂O₂. Moreover, olaparib failed to restore autophagy in SIRT1-depleted ARPE-19 cells, indicating that PARP1 inhibits autophagy through SIRT1 inhibition. Next, we further examined whether PARP1-induced autophagy impairment occurs in the retinas of dry AMD model mice. Histological analyses revealed that olaparib treatment protected mouse retinas against sodium iodate (SI) insult, but not in retinas cotreated with SI and wortmannin, an autophagy inhibitor. Collectively, our data demonstrate that PARP1-dependent inhibition of SIRT1 activity impedes autophagic survival of RPE cells, leading to retinal degeneration during dry AMD pathogenesis.

• Molecules and Cells
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• 제46권7호
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• pp.420-429
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• 2023

Age-related macular degeneration (AMD) is one of the leading causes of blindness in elderly individuals. However, the currently used intravitreal injections of anti-vascular endothelial growth factor are invasive, and repetitive injections are also accompanied by a risk of intraocular infection. The pathogenic mechanism of AMD is still not completely understood, but a multifactorial mechanism that combines genetic predisposition and environmental factors, including cellular senescence, has been suggested. Cellular senescence refers to the accumulation of cells that stop dividing due to the presence of free radicals and DNA damage. Characteristics of senescent cells include nuclear hypertrophy, increased levels of cell cycle inhibitors such as p16 and p21, and resistance to apoptosis. Senolytic drugs remove senescent cells by targeting the main characteristics of these cells. One of the senolytic drugs, ABT-263, which inhibits the antiapoptotic functions of Bcl-2 and Bcl-xL, may be a new treatment for AMD patients because it targets senescent retinal pigment epithelium (RPE) cells. We proved that it selectively kills doxorubicin (Dox)-induced senescent ARPE-19 cells by activating apoptosis. By removing senescent cells, the expression of inflammatory cytokines was reduced, and the proliferation of the remaining cells was increased. When ABT-263 was orally administered to the mouse model of senescent RPE cells induced by Dox, we confirmed that senescent RPE cells were selectively removed and retinal degeneration was alleviated. Therefore, we suggest that ABT-263, which removes senescent RPE cells through its senolytic effect, has the potential to be the first orally administered senolytic drug for the treatment of AMD.

• Molecules and Cells
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• 제46권11호
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• pp.675-687
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• 2023

Accumulation of pathogenic amyloid-β disrupts the tight junction of retinal pigment epithelium (RPE), one of its senescence-like structural alterations. In the clearance of amyloid-β, the autophagy-lysosome pathway plays the crucial role. In this context, mammalian target of rapamycin (mTOR) inhibits the process of autophagy and lysosomal degradation, acting as a potential therapeutic target for age-associated disorders. However, efficacy of targeting mTOR to treat age-related macular degeneration remains largely elusive. Here, we validated the therapeutic efficacy of the mTOR inhibitors, Torin and PP242, in clearing amyloid-β by inducing the autophagy-lysosome pathway in a mouse model with pathogenic amyloid-β with tight junction disruption of RPE, which is evident in dry age-related macular degeneration. High concentration of amyloid-β oligomers induced autophagy-lysosome pathway impairment accompanied by the accumulation of p62 and decreased lysosomal activity in RPE cells. However, Torin and PP242 treatment restored the lysosomal activity via activation of LAMP2 and facilitated the clearance of amyloid-β in vitro and in vivo. Furthermore, clearance of amyloid-β by Torin and PP242 ameliorated the tight junction disruption of RPE in vivo. Overall, our findings suggest mTOR inhibition as a new therapeutic strategy for the restoration of tight junctions in age-related macular degeneration.

• Journal of Photoscience
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• 제7권3호
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• pp.115-121
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• 2000

Although there are high concentrations of zinc and taurine in ocular tissue, their exact role and correlation in the visual process are not clear. The purpose of present study was to clarity this point using electroretinogram (ERG) recording and spectrophotometer measurements before and after zinc and taurine treatment in bullfrog's eye. The optimal zinc concentration used in this study was 10$^{-2}$ M ZnCl$_2$120 ${mu}ell$/12$m\ell$ ringer solution while the optimal turine concentration was 10$^{-2}$ M taurine 12${mu}ell$/12$m\ell$ ringer solution. For the effects of zinc and taurine on the retinal function, the changes of ERG parameters (especially threshold and b-wave) and absorption spectra were observed before and after treatment. It is noteworthy that high concentrations of zinc and taurine present in the retinal pigment epithelium and the retina. Our results indicate that dark-adapted ERG threshold became elevated and the peak amplitude of b-wave was increased with zinc and taurine treatment. Furthermore there are some synergism effects between zinc and taurine as a result of co-treatment. In spectral scan, absorbance increment due to zinc and taurine treatment was shown over the whole range of spectral range (300-750 nm) with some differences in absorbance increment depending on the case of treatment. As the results of above we believe that zinc and taurine, which are abundant in the retinal pigment epithelium and the retina particularly, may be essential factors for visual process, have some synergism with each other and be required to improve the visual sensitivity during visual adaptation.

• 대한안과학회지
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• 제59권12호
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• pp.1190-1194
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• 2018

목적: 단안에 국소적으로 발생한 색소정맥옆망막맥락막 위축(pigmented paravenous retinochoroidal atrophy) 1예를 경험하여 이를 보고하고자 한다. 증례요약: 46세 여자 환자가 건강검진상에서 발견된 우안 망막 이상 소견으로 내원하였다. 나안시력은 양안 모두 1.0이었다. 세극등현미경으로 시행한 전안부 검사상 특이 이상 소견은 관찰되지 않았다. 안저검사상 우안의 상측 망막정맥을 따라 국소적으로 분포된 뼈 조각 모양의 색소침착이 동반된 망막맥락막위축 소견이 관찰되었다. 안저자가형광상 우안의 망막맥락막위축의 경계에서 다소 과형광을 보이면서 병변의 대부분은 저형광 소견을 보였다. 빛간섭단층촬영상 해당 병변은 망막색소상피층의 위축 및 뭉침이 혼재되어 있었고, 맥락막층에서 맥락막모세혈관층이 얇아진 소견을 보였다. 형광안저촬영상 병변 내 창문비침 및 형광차단이 혼재되어 관찰되었으며 이는 망막색소상피층의 위축 및 뭉침 부위와 일치하였다. 결론: 단안에 국소적으로 발생한 색소정맥옆망막맥락막 위축 1예를 경험하여 이를 국내 최초로 보고하는 바이다.

• Journal of Photoscience
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• 제7권4호
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• pp.161-167
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• 2000

Temperature-dependent electroretinogram responses were investigated in the dark adapted bullfrog eyes within the physiological temperature range 0-40$\^{C}$. In hypothermic process(25→0→25$\^{C}$), the amplitude of b-and c-wave decreased with lowering the temperature again. Both b-wave amplitude and threshold responses were maximal around 15$\^{C}$ during the temperature increment. Upon warming to room temperature again (25$\^{C}$), the b-wave amplitude was approximately doubled as compared to that of control without temperature changes. During the hyperthermic process (25→40→25$\^{C}$), however, the responses decreased with warming, and the wave amplitude failed to recover by cooling to 25$\^{C}$ again. As describe above, the recoveries of ERG in both processes show the striking difference. The hypothermia induces the amplification of the b-wave, that is, enhances the retinal function with the temperature recovery toward room temperature. While the hypertherima produces the decrease of the b-wave even though recovered to room temperature, which indicates an irreversible retina. The morphological alteration is shown both hypothermic and hyperthermic process, such as an appearance of large vacuoles and degenerating outer segments, more intense in hyperthermia, similar to light induced damage.

• Journal of Photoscience
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• 제9권2호
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• pp.430-432
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• 2002

The changes in expression of copper-zinc superoxide dismutase (CuZn-SOD), manganese superoxide dismutase (Mn-SOD) and glutathione peroxidase (GPX) in light-damaged rat retinas were examined. Sprague-Dawley rats (male, 6-weeks-old) were maintained on a cyclic photoperiod (12 hours light and 12 hours darkness) for 2 weeks. The illumination intensity during the light period was 80 lux. To induce light damage to the retina, a high-intensity illumination (3000-lux) was applied to the animals for 24 hours. After light exposure, the animals were returned to cyclic lighting. Eyes were enucleated 12 and 24 hours after light exposure started or 1,3, and 7 days after light exposure ended. Eyes were fixed and embedded in paraffin wax. Tissues were cut into 4${\mu}{\textrm}{m}$-thick sections. Sections were immunostained using antibody against CuZn-SOD, Mn-SOD, GPX and 8-hydroxy-deoxyguanocine (8-OHdG) as oxidative stress marker. 8-OHdG was observed in the outer nuclear layer (ONL) and retinal pigment epithelium (RPE) during light exposure. In light-damaged retinas CuZn-SOD labeling was up regulated in the ONL and RPE. Mn-SOD labeling was up regulated in rod inner segments (RIS) during light exposure and that in the RPE was up regulated after exposure. GPX labeling was observed in rod outer segments (ROS) during light exposure. GPX labeling was also observed in the RPE during and after light exposure. All three enzymes were observed in the outer retina, which suffered light damage, but occurred in defferent layers except within the RPE, in which case all three were expressed. These enzymes may play complementary roles as protective factors in light-damaged retinas.

• Molecules and Cells
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• 제41권4호
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• pp.257-263
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• 2018

Vertebrate organ development is accompanied by demarcation of tissue compartments, which grow coordinately with their neighbors. Hence, perturbing the coordinative growth of neighboring tissue compartments frequently results in organ malformation. The growth of tissue compartments is regulated by multiple intercellular and intracellular signaling pathways, including the Hippo signaling pathway that limits the growth of various organs. In the optic neuroepithelial continuum, which is partitioned into the retina, retinal pigment epithelium (RPE) and ciliary margin (CM) during eye development, the Hippo signaling activity operates differentially, as it does in many tissues. In this review, we summarize recent studies that have explored the relationship between the Hippo signaling pathway and growth of optic neuroepithelial compartments. We will focus particularly on the roles of a tumor suppressor, neurofibromin 2 (NF2), whose expression is not only dependent on compartment-specific transcription factors, but is also subject to regulation by a Hippo-Yap feedback signaling circuit.

• 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.