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Cigarette Smoke Extract Causes Injury in Primary Retinal Ganglion Cells via Apoptosis and Autophagy

 Kwanghyun Lee  ;  Samin Hong  ;  Gong Je Seong  ;  Chan Yun Kim 
 CURRENT EYE RESEARCH, Vol.41(10) : 1367-1372, 2016 
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AIMS: To investigate whether tobacco smoke directly injures retinal ganglion cells (RGCs) and to evaluate the mechanisms of cell death. METHODS: Primary rat RGCs were harvested from 3- or 4-day-old newborn rats and exposed to cigarette smoke extract (CSE). Cell viability was determined by adenosine 5'-triphosphate (ATP) assay. Apoptosis was evaluated by terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) nick-end labeling (TUNEL) and real-time reverse transcription polymerase chain reaction (RT-PCR) for Bcl-2 family. Autophagy was also assessed by Western immunoblots for light chain (LC) 3B. RESULTS: When the primary RGCs were exposed to CSE for 2 h, cell viability decreased in a dose-dependent manner, as measured by ATP assay. In the presence of 0.05% CSE, the RGC viability was 77.68% ± 7.60% compared to the control cells; in the presence of 1.0% CSE, viability was 47.48% ± 2.56% of the control cells. As determined by TUNEL, CSE increased the apoptotic RGCs in a dose-dependent manner. In the presence of 0.05% CSE, the apoptosis was 26.55% ± 1.97% of the control cells; in the presence of 2.5% CSE, it was 41.07% ± 3.75% of the control cells. When apoptosis was evaluated using real-time RT-PCR, exposure to 0.05% CSE resulted in significantly increased expression of Bad, Bax, Bcl-2, and Bcl-XL mRNA. When autophagy was assessed by Western immunoblots, exposure to 0.05% CSE significantly increased the expression of LC3B II. CONCLUSIONS: Our data suggest that CSE directly injures primary RGCs, and both cell death mechanisms of apoptosis and autophagy seem to be related to this CSE-induced RGC damage.
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1. College of Medicine (의과대학) > Dept. of Ophthalmology (안과학교실) > 1. Journal Papers
Yonsei Authors
Kim, Chan Yun(김찬윤) ORCID logo https://orcid.org/0000-0002-8373-9999
Seong, Gong Je(성공제) ORCID logo https://orcid.org/0000-0002-5456-4296
Lee, Kwanghyun(이광현) ORCID logo https://orcid.org/0000-0002-1587-3387
Hong, Sa Min(홍사민)
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