Protective effects of 17β-estradiol on Benzo(e)pyrene[B(e)P]-induced toxicity in ARPE-19 cells

• Main Authors: Ashish U Sapkal, Sonali Nashine, Saffar Mansoor, Vishal R Sharma, Claudio A Ramirez, Rafael Z Migon, Navin K Gupta, Marilyn Chwa, Baruch D Kuppermann, Cristina Kenney
• Format: Article
• Language: English
• Published: Knowledge E 2018-01-01
• Series: Journal of Ophthalmic & Vision Research
• Subjects: 17β-Estradiol; Benzo(e)pyrene; Age-Related Macular Degeneration; Apoptosis; Retinal Pigment Epithelial Cells; Smoking
• Online Access: http://www.jovr.org/article.asp?issn=2008-322X;year=2018;volume=13;issue=4;spage=419;epage=425;aulast=Sapkal

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.