The Effect of A2E on the Uptake and Release of Calcium in the Lysosomes and Mitochondria of Human RPE Cells Exposed to Blue Light
We aimed to explore the effect of N-retinylidene-N-retinylethanolamine (A2E) on the uptake and release of calcium in lysosomes and mitochondria by establishing a model of human retinal pigment epithelial (RPE) cell injury induced by exposure to blue light. Primary human RPE cells were cultured from...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2021-01-01
|
| Series: | Journal of Ophthalmology |
| Online Access: | http://dx.doi.org/10.1155/2021/5586659 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832560628106002432 |
|---|---|
| author | Mao-Mei Luo Lin Chen Shu Wang Chun Zeng De-Zhi Li YeGe Bi Long-Qian Liu Shan-Jun Cai |
| author_facet | Mao-Mei Luo Lin Chen Shu Wang Chun Zeng De-Zhi Li YeGe Bi Long-Qian Liu Shan-Jun Cai |
| author_sort | Mao-Mei Luo |
| collection | DOAJ |
| description | We aimed to explore the effect of N-retinylidene-N-retinylethanolamine (A2E) on the uptake and release of calcium in lysosomes and mitochondria by establishing a model of human retinal pigment epithelial (RPE) cell injury induced by exposure to blue light. Primary human RPE cells were cultured from passages 4 to 6 and exposed to blue light at an intensity of 2000 ± 500 lux for 6 hours. After blue light exposure, the culture was maintained for 24 hours. A2E at a final concentration of 25 μM was added to the culture 2 hours before light exposure, and nifedipine at a final concentration of 10−4 M was added 1 hour before light exposure. The levels of Ca2+ in the cytosol (CaTM/2AM), mitochondria (Rhod/2AM), and lysosomes (LysoTracker Red and Fluo-3/AM) were determined. In order to measure the calcium levels in the different organelles, RPE were imaged using a laser scanning confocal microscope. Moreover, changes in the mitochondrial membrane potential were detected by flow cytometry analysis of JC-1-stained cells. The obtained results revealed that blue light illumination increased the calcium fluorescence intensity in the cytoplasm, mitochondria, and lysosomes of human RPE cells when compared with the control cells (P<0.05). After A2E treatment, the fluorescence intensity of the calcium in the cytoplasm was further increased (P<0.05), while that in the mitochondria and lysosomes decreased (P<0.05). In addition, we observed that nifedipine reduced the fluorescence intensity of calcium in the RPE cells. Our results also showed that the mitochondrial membrane potential in the RPE treated with blue light and A2E was lower than that in the control, blue light, and A2E-treated cells (P<0.05). Blue light increased calcium levels in the cytoplasm, lysosomes, and mitochondria of RPE cells. A2E damages the lysosomal and mitochondrial membranes, resulting in calcium release into the cytoplasm. Finally, our results demonstrated that both blue light and A2E treatments reduced mitochondrial membrane potential, increasing cytosolic Ca2+ levels, which can contribute to the activation of RPE death. |
| format | Article |
| id | doaj-art-d6f7b719230c455ca8be56a7b4afd8ff |
| institution | Kabale University |
| issn | 2090-004X 2090-0058 |
| language | English |
| publishDate | 2021-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Ophthalmology |
| spelling | doaj-art-d6f7b719230c455ca8be56a7b4afd8ff2025-02-03T01:27:05ZengWileyJournal of Ophthalmology2090-004X2090-00582021-01-01202110.1155/2021/55866595586659The Effect of A2E on the Uptake and Release of Calcium in the Lysosomes and Mitochondria of Human RPE Cells Exposed to Blue LightMao-Mei Luo0Lin Chen1Shu Wang2Chun Zeng3De-Zhi Li4YeGe Bi5Long-Qian Liu6Shan-Jun Cai7Department of Ophthalmology, Affiliated Hospital of Zunyi Medical University, Zunyi, Guizhou, ChinaDepartment of Ophthalmology, Affiliated Hospital of Zunyi Medical University, Zunyi, Guizhou, ChinaDepartment of Ophthalmology, Affiliated Hospital of Zunyi Medical University, Zunyi, Guizhou, ChinaDepartment of Ophthalmology, Affiliated Hospital of Zunyi Medical University, Zunyi, Guizhou, ChinaDepartment of Ophthalmology, Affiliated Hospital of Zunyi Medical University, Zunyi, Guizhou, ChinaDepartment of Ophthalmology, Affiliated Hospital of Zunyi Medical University, Zunyi, Guizhou, ChinaDepartment of Ophthalmology, West China Hospital, Sichuan University, Chengdu, Sichuan, ChinaDepartment of Ophthalmology, Affiliated Hospital of Zunyi Medical University, Zunyi, Guizhou, ChinaWe aimed to explore the effect of N-retinylidene-N-retinylethanolamine (A2E) on the uptake and release of calcium in lysosomes and mitochondria by establishing a model of human retinal pigment epithelial (RPE) cell injury induced by exposure to blue light. Primary human RPE cells were cultured from passages 4 to 6 and exposed to blue light at an intensity of 2000 ± 500 lux for 6 hours. After blue light exposure, the culture was maintained for 24 hours. A2E at a final concentration of 25 μM was added to the culture 2 hours before light exposure, and nifedipine at a final concentration of 10−4 M was added 1 hour before light exposure. The levels of Ca2+ in the cytosol (CaTM/2AM), mitochondria (Rhod/2AM), and lysosomes (LysoTracker Red and Fluo-3/AM) were determined. In order to measure the calcium levels in the different organelles, RPE were imaged using a laser scanning confocal microscope. Moreover, changes in the mitochondrial membrane potential were detected by flow cytometry analysis of JC-1-stained cells. The obtained results revealed that blue light illumination increased the calcium fluorescence intensity in the cytoplasm, mitochondria, and lysosomes of human RPE cells when compared with the control cells (P<0.05). After A2E treatment, the fluorescence intensity of the calcium in the cytoplasm was further increased (P<0.05), while that in the mitochondria and lysosomes decreased (P<0.05). In addition, we observed that nifedipine reduced the fluorescence intensity of calcium in the RPE cells. Our results also showed that the mitochondrial membrane potential in the RPE treated with blue light and A2E was lower than that in the control, blue light, and A2E-treated cells (P<0.05). Blue light increased calcium levels in the cytoplasm, lysosomes, and mitochondria of RPE cells. A2E damages the lysosomal and mitochondrial membranes, resulting in calcium release into the cytoplasm. Finally, our results demonstrated that both blue light and A2E treatments reduced mitochondrial membrane potential, increasing cytosolic Ca2+ levels, which can contribute to the activation of RPE death.http://dx.doi.org/10.1155/2021/5586659 |
| spellingShingle | Mao-Mei Luo Lin Chen Shu Wang Chun Zeng De-Zhi Li YeGe Bi Long-Qian Liu Shan-Jun Cai The Effect of A2E on the Uptake and Release of Calcium in the Lysosomes and Mitochondria of Human RPE Cells Exposed to Blue Light Journal of Ophthalmology |
| title | The Effect of A2E on the Uptake and Release of Calcium in the Lysosomes and Mitochondria of Human RPE Cells Exposed to Blue Light |
| title_full | The Effect of A2E on the Uptake and Release of Calcium in the Lysosomes and Mitochondria of Human RPE Cells Exposed to Blue Light |
| title_fullStr | The Effect of A2E on the Uptake and Release of Calcium in the Lysosomes and Mitochondria of Human RPE Cells Exposed to Blue Light |
| title_full_unstemmed | The Effect of A2E on the Uptake and Release of Calcium in the Lysosomes and Mitochondria of Human RPE Cells Exposed to Blue Light |
| title_short | The Effect of A2E on the Uptake and Release of Calcium in the Lysosomes and Mitochondria of Human RPE Cells Exposed to Blue Light |
| title_sort | effect of a2e on the uptake and release of calcium in the lysosomes and mitochondria of human rpe cells exposed to blue light |
| url | http://dx.doi.org/10.1155/2021/5586659 |
| work_keys_str_mv | AT maomeiluo theeffectofa2eontheuptakeandreleaseofcalciuminthelysosomesandmitochondriaofhumanrpecellsexposedtobluelight AT linchen theeffectofa2eontheuptakeandreleaseofcalciuminthelysosomesandmitochondriaofhumanrpecellsexposedtobluelight AT shuwang theeffectofa2eontheuptakeandreleaseofcalciuminthelysosomesandmitochondriaofhumanrpecellsexposedtobluelight AT chunzeng theeffectofa2eontheuptakeandreleaseofcalciuminthelysosomesandmitochondriaofhumanrpecellsexposedtobluelight AT dezhili theeffectofa2eontheuptakeandreleaseofcalciuminthelysosomesandmitochondriaofhumanrpecellsexposedtobluelight AT yegebi theeffectofa2eontheuptakeandreleaseofcalciuminthelysosomesandmitochondriaofhumanrpecellsexposedtobluelight AT longqianliu theeffectofa2eontheuptakeandreleaseofcalciuminthelysosomesandmitochondriaofhumanrpecellsexposedtobluelight AT shanjuncai theeffectofa2eontheuptakeandreleaseofcalciuminthelysosomesandmitochondriaofhumanrpecellsexposedtobluelight AT maomeiluo effectofa2eontheuptakeandreleaseofcalciuminthelysosomesandmitochondriaofhumanrpecellsexposedtobluelight AT linchen effectofa2eontheuptakeandreleaseofcalciuminthelysosomesandmitochondriaofhumanrpecellsexposedtobluelight AT shuwang effectofa2eontheuptakeandreleaseofcalciuminthelysosomesandmitochondriaofhumanrpecellsexposedtobluelight AT chunzeng effectofa2eontheuptakeandreleaseofcalciuminthelysosomesandmitochondriaofhumanrpecellsexposedtobluelight AT dezhili effectofa2eontheuptakeandreleaseofcalciuminthelysosomesandmitochondriaofhumanrpecellsexposedtobluelight AT yegebi effectofa2eontheuptakeandreleaseofcalciuminthelysosomesandmitochondriaofhumanrpecellsexposedtobluelight AT longqianliu effectofa2eontheuptakeandreleaseofcalciuminthelysosomesandmitochondriaofhumanrpecellsexposedtobluelight AT shanjuncai effectofa2eontheuptakeandreleaseofcalciuminthelysosomesandmitochondriaofhumanrpecellsexposedtobluelight |