Author/Authors :
Sapkal, Ashish U Department of Ophthalmology - Gavin Herbert Eye Institute - University of California - Irvine - CA, USA , Nashine, Sonali Department of Ophthalmology - Gavin Herbert Eye Institute - University of California - Irvine - CA, USA , Mansoor, Saffar Department of Ophthalmology - Gavin Herbert Eye Institute - University of California - Irvine - CA, USA , Sharma, Vishal R Department of Ophthalmology - Gavin Herbert Eye Institute - University of California - Irvine - CA, USA , Ramirez, Claudio A Department of Ophthalmology - Gavin Herbert Eye Institute - University of California - Irvine - CA, USA , Migon, Rafael Z Department of Ophthalmology - Gavin Herbert Eye Institute - University of California - Irvine - CA, USA , Gupta, Navin K Department of Ophthalmology - Gavin Herbert Eye Institute - University of California - Irvine - CA, USA , Chwa, Marilyn Department of Ophthalmology - Gavin Herbert Eye Institute - University of California - Irvine - CA, USA , Kuppermann, Baruch D Department of Ophthalmology - Gavin Herbert Eye Institute - University of California - Irvine - CA, USA , Kenney, M. Cristina Department of Ophthalmology - Gavin Herbert Eye Institute - University of California - Irvine - CA, USA
Abstract :
Purpose: The aim of this study was to examine the effect of 17β‑estradiol on Benzo(e)pyrene [B(e)P]‑induced
toxicity in ARPE‑19 cells.
Methods: We pretreated ARPE‑19 cells with 20 nM and 40 nM 17β‑estradiol for 6 hours, followed by addition
of 300 μM B(e)P for additional 24 hours. Cell viability was measured using Trypan blue dye‑exclusion
assay. JC‑1 assay was performed to measure mitochondrial membrane potential (ΔΨm). For a quantitative
estimation of cell death, apoptotic markers such as caspase‑3/7, caspase‑9, and caspase‑12 were measured.
Results: Our results demonstrated that when treated with B(e)P, the viability and ΔΨm of ARPE‑19 cells
declined by 25% and 63%, respectively (P < 0.05). However, pretreating with 17β‑estradiol increased the
viability of ARPE‑19 cells by 21% (20 nM) and 10% (40 nM) (P < 0.05). Furthermore, the significantly reduced
ΔΨm in βE+B(e)P treated cells ARPE‑19 cells was restored by pre‑treatment with 17β‑estradiol‑ ΔΨm was
increased by 177% (20 nM) and 158% (40 nM) (P < 0.05). We further observed a significant up‑regulation in
the activity of Caspases‑3/7, ‑9, and ‑12 in B(e)P‑treated ARPE‑19 cells. However, 17β‑estradiol treatment
significantly reduced the activity of all apoptotic markers (P < 0.05).
Conclusion: In conclusion, our results demonstrate that 17β‑estradiol protects ARPE‑19 cells against B(e)
P‑induced toxicity by decreasing apoptosis, preventing cell death, and restoring mitochondrial membrane
potential.
Keywords :
17β‑Estradiol , Benzo(e)pyrene , Age‑Related Macular Degeneration , Apoptosis , Retinal Pigment Epithelial Cells , Smoking
