Contribution of Calpain and Caspases to Cell Death in Cultured Monkey RPE Cells
| Autor: | Thomas R. Shearer, Katherine B. Hammond, Emi Nakajima, Mitsuyoshi Azuma, Masayuki Hirata |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Proteases Programmed cell death Cell Survival Immunoblotting Caspase 3 non-human primate Apoptosis Retinal Pigment Epithelium AMD A2E 03 medical and health sciences Retinoids 0302 clinical medicine medicine Animals Microscopy Phase-Contrast Enzyme Inhibitors Caspase Cells Cultured Retinal pigment epithelium biology Chemistry hypoxia Calpain Biochemistry and Molecular Biology Hypoxia (medical) Caspase Inhibitors Macaca mulatta eye diseases Cell Hypoxia 3. Good health Cell biology 030104 developmental biology medicine.anatomical_structure Proto-Oncogene Proteins c-bcl-2 Caspases 030221 ophthalmology & optometry biology.protein sense organs medicine.symptom |
| Zdroj: | Investigative Ophthalmology & Visual Science |
| ISSN: | 1552-5783 0146-0404 |
| Popis: | Purpose AMD is the leading cause of human vision loss after 65 years of age. Several mechanisms have been proposed: (1) age-related failure of the choroidal vasculature leads to loss of RPE; (2) RPE dysfunctions due to accumulation of phagocytized, but unreleased A2E (N-retinylidene-N-retinylethanolamine); (3) zinc deficiency activation of calpain and caspase proteases, leading to cell death. The purpose of the present study is to compare activation of calpain and caspase in monkey RPE cells cultured under hypoxia or with A2E. Methods Monkey primary RPE cells were cultured under hypoxic conditions in a Gaspak pouch or cultured with synthetic A2E. Immunoblotting was used to detect activation of calpain and caspase. Calpain inhibitor, SNJ-1945, and pan-caspase inhibitor, z-VAD-fmk, were used to confirm activation of the proteases. Results (1) Hypoxia and A2E each decreased viability of RPE cells in a time-dependent manner. (2) Incubation under hypoxia alone induced activation of calpain, but not caspases. SNJ-1945 inhibited calpain activation, but z-VAD-fmk did not. (3) Incubation with A2E alone induced activation of calpain, caspase-9, and caspase-3. SNJ-1945 inhibited calpain activation. z-VAD-fmk inhibited caspase activation, suggesting no interaction between calpain and caspases. Conclusions Hypoxia activated the calpain pathway, while A2E activated both calpain and caspase pathways in monkey RPE cells. Such knowledge may be utilized in the treatment of AMD if inhibitor drugs against calpain and/or caspase are used to prevent RPE dysfunction caused by hypoxia or A2E. |
| Databáze: | OpenAIRE |
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