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Long-term blue light exposure induces RGC-5 cell death in vitro: involvement of mitochondria-dependent apoptosis, oxidative stress, and MAPK signaling pathways

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Abstract

The mechanism of blue light-induced retinal ganglion cell (RGC) injury is poorly understood. In this study, we established a patented light-emitting diode-based system to study the effects of long-term blue light exposure under culture conditions on RGC-5 cells. Long-term blue light exposure significantly reduced cell viability in a time-dependent manner and induced apoptosis and necrosis in RGC-5 cells. Long-term blue light exposure marked an increase in the expression of Bax and active Caspase-3 (p17), which was accompanied by Bcl-2 down-regulation, and displayed features of the mitochondria-dependent apoptosis pathway. Blue light exposure also increased the generation of reactive oxygen species (ROS), and was a strong inducer of ROS-sensitive protein nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) expression. Moreover, blue light exposure constitutively activated p38 mitogen-activated protein kinases and c-Jun NH2-terminal kinase (JNK), as well as induced the phosphorylation of extracellular signal-regulated kinase in the early phase, in blue light-exposed RGC-5 cells. The protein expression of c-jun and c-fos was further enhanced after RGC-5 cells were exposed to blue light. Taken together, these findings indicated that blue light induced RGC-5 cell line death in dependence upon exposure duration. The potential mechanisms for this phenomenon might be via activated mitochondria-dependent apoptosis, increased ROS production and protein expressions of Nrf2 and HO-1, and activated JNK/p38 MAPK signaling pathways.

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Abbreviations

AMD:

Age related macular degeneration

RPE:

Retinal pigment epithelium

RGC:

Retinal ganglion cell

ROS:

Reactive oxygen species

AIF:

Apoptosis-inducing factor

MAPK:

Mitogen-activated protein kinase

ERK:

Extracellular signal-regulated protein kinase

JNK:

c-Jun NH2-terminal kinase

LED:

Light-emitting diode

DMEM:

Dulbecco’s modified Eagle’s media

EDTA:

Ethylene diamine tetra acetic acid

PBS:

Phosphate buffer solution

FBS:

Fetal bovine serum

CCK-8:

Cell counting kit-8

PI:

Propidium iodide

H2DCFDA:

2′–7′ Dichlorofluorescence diacetate

FITC:

Fluorescein isothiocyanate

RIPA:

Radioimmunoprecipitation

BCA:

Bicinchoninic acid

Nrf2:

Nuclear factor erythroid 2-related factor 2

HO-1:

Heme oxygenase-1

HRP:

Horseradish peroxidase

SD:

Standard deviation

TUNEL:

Terminal-deoxynucleoitidyl transferase mediated nick end labeling

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Acknowledgments

The research was supported by National Natural Science Foundation of China (No. 81170888).

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Authors and Affiliations

  1. Department of Ophthalmology, Peking University Third Hospital, 49 North Garden Road, Haidian District, Beijing, 100191, China

    Chen Huang, Pei Zhang, Wei Wang, Yongsheng Xu, Minshu Wang, Xiaoyong Chen & Xuran Dong

  2. Medical Research Center, Peking University Third Hospital, 49 North Garden Road, Haidian District, Beijing, 100191, China

    Chen Huang

  3. Clinical Stem Cell Center, Peking University Third Hospital, 49 North Garden Road, Haidian District, Beijing, 100191, China

    Yongsheng Xu

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  1. Chen Huang
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Correspondence to Wei Wang.

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Huang, C., Zhang, P., Wang, W. et al. Long-term blue light exposure induces RGC-5 cell death in vitro: involvement of mitochondria-dependent apoptosis, oxidative stress, and MAPK signaling pathways. Apoptosis 19, 922–932 (2014). https://doi.org/10.1007/s10495-014-0983-2

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