[논문]Neuroprotective effect of Aster yomena (Kitam.) Honda against hydrogen peroxide-induced oxidative stress in SH-SY5Y cells

Kim, Min Jeong; Kim, Ji Hyun; Lee, Sanghyun; Cho, Eun Ju; Kim, Hyun Young

doi:10.3839/jabc.2020.038

Neuroprotective effect of Aster yomena (Kitam.) Honda against hydrogen peroxide-induced oxidative stress in SH-SY5Y cells 원문보기

Journal of applied biological chemistry, v.63 no.3, 2020년, pp.283 - 290

Kim, Min Jeong (Department of Food Science and Nutrition, Pusan National University) , Kim, Ji Hyun (Department of Food Science, Gyeongnam National University of Science and Technology) , Lee, Sanghyun (Department of Plant Science and Technology, Chung-Ang University) , Cho, Eun Ju (Department of Food Science and Nutrition, Pusan National University) , Kim, Hyun Young (Department of Food Science, Gyeongnam National University of Science and Technology)

Abstract ▼ AI-Helper

Oxidative stress is one of the contributors of neurodegenerative disorders including Alzheimer's disease. According to previous studies, Aster yomena (Kitam.) Honda (AY) possesses variable pharmacological activities including anti-coagulant and anti-obesity effect. In this study, we aimed to determine the neuroprotective effect of ethyl acetate fraction from Aster yomena (Kitam.) Honda (EFAY) against oxidative stress. Therefore, we carried out 3-(4,5-dimethylthiazol-2-yl)-2,3-diphenyl tetrazolium bromide, lactate dehydrogenase (LDH), and 2',7'-dichlorofluorescin diacetate assays in SH-SY5Y neuronal cells treated with hydrogen peroxide (H2O2). H2O2-treated control cells exhibited reduced viability of cells, and increased LDH release and reactive oxygen species (ROS) production compared to normal cells. However, treatment with EFAY restored the cell viability and inhibited LDH release and ROS production. To investigate the underlying mechanisms by which EFAY attenuated neuronal oxidative damage, we measured protein expressions using Western blot analysis. Consequently, it was observed that EFAY down-regulated cyclooxygenase-2 and interleukin-1β protein expressions in H2O2-treated SH-SY5Y cells that mediated inflammatory reaction. In addition, apoptosis-related proteins including B-cell lymphoma-2-associated X protein/B-cell lymphoma-2 ratio, cleaved caspase-9, and cleaved-poly (ADP-ribose) polymerase protein expressions were suppressed when H2O2-treated cells were exposed to EFAY. Our results indicate that EFAY ameliorated H2O2-induced neuronal damage by regulating inflammation and apoptosis. Altogether, AY could be potential therapeutic agent for neurodegenerative diseases.

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가설 설정

Based on the obtained results, the inhibitory effect of EFAY on inflammation and apoptosis pathways is apparent. In this study, we investigated neuroprotective effect of AY via inflammatory and apoptosis pathway. According to previous study, AY inhibited HO-induced growth inhibition in RAW 264.

제안 방법

The cell viability was measured to determine the protective effect of EFAY on H₂O₂-treated SH-SY5Y cells by performing MTT assay. As illustrated in Fig.

대상 데이터

The SH-SY5Y cell line was obtained from KCLB (Korean Cell Line Bank, Seoul, Korea). SH-SY5Y cells were maintained in DMEM supplemented with 10% FBS and 100 units/mL penicillin-streptomycin.

데이터처리

The significance of data was determined by oneway analysis of variance (ANOVA) and Duncan’s multiple range test for multiple comparisons.

성능/효과

Previous studies demonstrated that treatment with H₂O₂ led to the formation of apoptotic features such as decreased Bax/Bcl-2 ratio, activation of the caspase cascade, and DNA fragmentation in SH-SY5Y cells[31,53,54]. Our results showed that H₂O₂-mediated neuronal apoptosis was activated in SH-SY5Y cells by up-regulating Bax/Bcl-2 ratio, whereas EFAY treatment down-regulated Bax/Bcl-2 ratio. Furthermore, up-regulated protein expressions of cleaved caspase-9 and cleaved PARP were observed subsequent to treatment of SH-SY5Y cells with H₂O₂.
99%. The results demonstrated that ROS production caused by H₂O₂ was blocked by EFAY with subsequent reduction in neuronal cell death.
However, treatment with EFAY significantly down-regulated Bax/Bcl-2 ratio and up-regulated cleaved caspase-9 and cleaved PARP protein levels. These results indicated that the treatment of H₂O₂-induced damaged SH-SY5Y cells with EFAY suppressed apoptosis pathway by modulating Bax/Bac-2 ratio, cleaved caspase-9, and cleaved PARP.

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