Hydrogen Peroxide-Induced Cell Death in a Human Retinal Pigment Epithelial Cell Line, ARPE-19

Kim, Min Ho; Chung, Jae Eun; Yang, Ji Wook; Chung, Sang Moon; Kwag, No Hoon; Yoo, Jin Seong

doi:10.3341/kjo.2003.17.1.19

Cited by 66 publications

(48 citation statements)

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“…2). The absorbance of H 2 O 2 at 800 µmol/L by MTT assay was significantly lower than that of the control group (P<0.05), indicating that the dosage of H 2 O 2 to induce a half-value of MC3T3-E1 apoptosis was around 800 µmol/L, which is close to the previously reported value (600 µmol/L) by Kim et al (2003).…”

Section: Mc3t3-e1 Apoptosis Induced By H 2 O 2 At Various Concentrationssupporting

confidence: 80%

Eucommia ulmoides Oliv. antagonizes H2O2-induced rat osteoblastic MC3T3-E1 apoptosis by inhibiting expressions of caspases 3, 6, 7, and 9

Lin

Fan

Mehl

et al. 2011

J. Zhejiang Univ. Sci. B

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Eucommia ulmoides Oliv. (EuO), also known as Duzhong, native to China, has been reported to have antioxidative function, but its cellular mechanism is not fully examined yet. We investigated inhibitory effects of EuO leaf ethanol extracts on H 2 O 2 -induced apoptosis in rat osteoblastic MC3T3-E1 cells and underlying mechanisms. Locally-grown Duzhong leaves were extracted with ethanol. MC3T3-E1 cells were treated with EuO (6.25, 12.5, 25, 50, and 100 µg/ml) for 24 h, and then H 2 O 2 (800 µmol/L) for an additional 24 h. Cell survival rate, percentage of apoptosis, and expressions of caspases 3, 6, 7, and 9 were examined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, microscopic analysis, Western blotting, and reverse transcription polymerase chain reaction (RT-PCR). The final EuO leaf ethanol extract powder was detected to contain caffeotannic acid at 58 mg/g and geniposide at 3.45 mg/g by high performance liquid chromatography (HPLC). EuO remarkably restrained cell oxidative damage and increased cell survival rate in a dose-dependent manner: 0 µg/ml, 0.21; 6.25 µg/ml, 0. 28; 12.5 µg/ml, 0.31; 25 µg/ml, 0.48; 50 µg/ml, 0.54; and 100 µg/ml, 0.66 (P<0.05), with the half-effective concentration being around 25 µg/ml. MTT results were confirmed by microscopic analysis. Western blotting and RT-PCR analyses showed that the expressions of caspases 3, 6, 7, and 9 were significantly decreased in the EuO-treated cells compared with the control (EuO-and H 2 O 2 -free) (P<0.05), with the half-effective concentration of EuO ranging from 12.5 to 25 µg/ml. We conclude that the ethanol-extracted EuO leaf extracts promoted the growth of MC3T3-E1 cells, and suppressed the H 2 O 2 -induced apoptosis in a rat MC3T3-E1 osteogenic cell model, likely due to the inhibition of caspases' activities. The results indicate that EuO is a potent antioxidant, which may contribute to its many cellular protective functions, including the promotion of bone growth.

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Section: Mc3t3-e1 Apoptosis Induced By H 2 O 2 At Various Concentrationssupporting

confidence: 80%

Eucommia ulmoides Oliv. antagonizes H2O2-induced rat osteoblastic MC3T3-E1 apoptosis by inhibiting expressions of caspases 3, 6, 7, and 9

Lin

Fan

Mehl

et al. 2011

J. Zhejiang Univ. Sci. B

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“…The eye is a highly oxidative stressed tissue. In normal human eyes, H 2 O 2 levels in the aqueous humor have been reported to be in the range of 14-31 mM, whereas under pathological conditions such as cataracts, H 2 O 2 levels can range from 33 to 500 mM in the aqueous humor [36][37][38] at this pathological range stimulating apoptosis in RPE cells in vitro and shown high numbers of apoptotic RPE cells in postmortem AMD eyes [23,39,40]. Because the implanted RPE will remain in the highly oxidative stressed environment of an AMD patient's eye, we simulated those pathological conditions with high dosages of H 2 O 2 to treat hESC-RPE cells in vitro and compared the resistance to the oxidative stress between the polarized and nonpolarized RPE cells.…”

Section: Discussionmentioning

confidence: 99%

Polarized Human Embryonic Stem Cell-Derived Retinal Pigment Epithelial Cell Monolayers Have Higher Resistance to Oxidative Stress-Induced Cell Death Than Nonpolarized Cultures

Hsiung

Zhu

Hinton

2014

Stem Cells Translational Medicine

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Oxidative stress-mediated injury to the retinal pigment epithelium (RPE) is a major factor involved in the pathogenesis of age-related macular degeneration (AMD), the leading cause of blindness in the elderly. Human embryonic stem cell (hESC)-derived RPE cells are currently being evaluated for their potential for cell therapy in AMD patients through subretinal injection of cells in suspension and subretinal placement as a polarized monolayer. To gain an understanding of how transplanted RPE cells will respond to the highly oxidatively stressed environment of an AMD patient eye, we compared the survival of polarized and nonpolarized RPE cultures following oxidative stress treatment. Polarized, nonpolarized/confluent, nonpolarized/subconfluent hESC-RPE cells were treated with H 2 O 2 . Terminal deoxynucleotidyl transferase dUTP nick end labeling stains revealed the highest amount of cell death in subconfluent hESC-RPE cells and little cell death in polarized hESC-RPE cells with H 2 O 2 treatment. There were higher levels of proapoptotic factors (phosphorylated p38, phosphorylated c-Jun NH 2 -terminal kinase, Bax, and cleaved caspase 3 fragments) in treated nonpolarized RPE-particularly subconfluent cells-relative to polarized cells. On the other hand, polarized RPE cells had constitutively higher levels of cell survival and antiapoptotic signaling factors such as p-Akt and Bcl-2, as well as antioxidants superoxide dismutase 1 and catalase relative to nonpolarized cells, that possibly contributed to polarized cells' higher tolerance to oxidative stress compared with nonpolarized RPE cells. Subconfluent cells were particularly sensitive to oxidative stress-induced apoptosis. These results suggest that implantation of polarized hESC-RPE monolayers for treating AMD patients with geographic atrophy should have better survival than injections of hESC-RPE cells in suspension. STEM CELLS TRANSLATIONAL MEDICINE 2015;4:10-20

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“…The mechanism of RPE cell death in response to oxidative stress and in GA has been controversial. Apoptosis was suggested as a major mechanism of RPE cell death, even though several studies suggested necrosis as mechanism of RPE cell death in vitro (9,10) and in vivo (11,12). Necrosis used to be considered a passive and unregulated form of cell death.…”

mentioning

confidence: 99%

Gossypol Acetic Acid Prevents Oxidative Stress-Induced Retinal Pigment Epithelial Necrosis by Regulating the FoxO3/Sestrin2 Pathway

Hanus

Zhang

Chen

et al. 2015

Molecular and Cellular Biology

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The late stage of dry age-related macular degeneration (AMD), or geographic atrophy (GA), is characterized by extensive retinal pigment epithelial (RPE) cell death, and a cure is not available currently. We have recently demonstrated that RPE cells die from necrosis in response to oxidative stress, providing a potential novel mechanism for RPE death in AMD. In this study, we screened U.S. Food and Drug Administration-approved natural compounds and identified gossypol acetic acid (GAA) as a potent inhibitor of oxidative stress-induced RPE cell death. GAA induces antioxidative response and inhibits accumulation of excessive reactive oxygen species in cells, through which it prevents the activation of intrinsic necrotic pathway in response to oxidative stress. Sestrin2 (SESN2) is found to mediate GAA function in antioxidative response and RPE survival upon oxidative stress. Moreover, Forkhead box O3 transcription factor (FoxO3) is further found to be required for GAA-mediated SESN2 expression and RPE survival. Mechanistically, GAA promotes FoxO3 nuclear translocation and binding to the SESN2 enhancer, which in turn increases its transcriptional activity. Taken together, we have identified GAA as a potent inhibitor of oxidative stress-induced RPE necrosis by regulating the FoxO3/SESN2 pathway. This study may have significant implications in the therapeutics of age-related diseases, especially GA.A ge-related macular degeneration (AMD) is the leading cause of severe vision loss in people aged over 50, and its prevalence increases exponentially in people over the age of 70 (1). Currently, it is estimated that 1.75 million individuals suffer from this disease in the United States, and 7 million are said to be "at risk" (2). There are two types of AMD, the "dry" and "wet" forms, respectively. Dry AMD is a chronic disease that usually causes some degree of visual impairment and sometimes progresses to severe blindness. Dry AMD accounts for 90% of AMD cases and is currently without treatment available. The late stage of dry AMD, which is also knows as geographic atrophy (GA), is characterized by scattered or confluent areas of degeneration of retinal pigment epithelium (RPE) cells and the overlying photoreceptors that rely on the RPE for trophic support (3). AMD is a multifactorial disease with unclear etiology. Age is the most consistent risk factor associated with AMD, and genetic factors, oxidative stress, and inflammation also significantly contribute to AMD pathogenesis (4). Cigarette smoking, which induces systemic oxidative stress, has been proved to be a significant risk factor for AMD. Consistently, clinical studies have shown that the progression of AMD can be slowed with antioxidant vitamins and zinc supplements (5, 6). The retina is one of the highest oxygen-consuming tissues in the human body and, in particular, RPE is vulnerable to oxidative damage (7,8). The mechanism of RPE cell death in response to oxidative stress and in GA has been controversial. Apoptosis was suggested as a major mechanism of RPE cell ...

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Hydrogen Peroxide-Induced Cell Death in a Human Retinal Pigment Epithelial Cell Line, ARPE-19

Cited by 66 publications

References 1 publication

Eucommia ulmoides Oliv. antagonizes H2O2-induced rat osteoblastic MC3T3-E1 apoptosis by inhibiting expressions of caspases 3, 6, 7, and 9

Eucommia ulmoides Oliv. antagonizes H2O2-induced rat osteoblastic MC3T3-E1 apoptosis by inhibiting expressions of caspases 3, 6, 7, and 9

Polarized Human Embryonic Stem Cell-Derived Retinal Pigment Epithelial Cell Monolayers Have Higher Resistance to Oxidative Stress-Induced Cell Death Than Nonpolarized Cultures

Gossypol Acetic Acid Prevents Oxidative Stress-Induced Retinal Pigment Epithelial Necrosis by Regulating the FoxO3/Sestrin2 Pathway

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