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Insights on the involvement of (–)-epigallocatechin gallate in ER stress-mediated apoptosis in age-related macular degeneration

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Abstract

Endoplasmic reticulum (ER) stress-mediated apoptosis is a well-known factor in the pathogenesis of age-related macular degeneration (AMD). ER stress leads to accumulation of misfolded proteins, which in turn activates unfolded protein response (UPR) of the cell for its survival. The prolonged UPR of ER stress promotes cell death; however, the transition between adaptation and ER stress-induced apoptosis has not been clearly understood. Hence, the present study investigates the regulatory effect of (–)-epigallocatechin gallate (EGCG) on ER stress-induced by hydrogen peroxide (H2O2) and disturbance of calcium homeostasis by thapsigargin (TG) in mouse retinal pigment epithelial (MRPE) cells. The oxidant molecules influenced MRPE cells showed an increased level of intracellular calcium [Ca2+]i in ER and transferred to mitochondria through ER-mitochondrial tether site then increased ROS production. EGCG restores [Ca2+]i homeostasis by decreasing ROS production through inhibition of prohibitin1 which regulate ER-mitochondrial tether site and inhibit apoptosis. Effect of EGCG on ER stress-mediated apoptosis was elucidated by exploring the UPR signalling pathways. EGCG downregulated GRP78, CHOP, PERK, ERO1α, IRE1α, cleaved PARP, cleaved caspase 3, caspase 12 and upregulated expression of calnexinin MRPE cells. In addition to this, inhibition of apoptosis by EGCG was also confirmed with expression of proteins Akt, PTEN and GSK3β. MRPE cells with EGCG upregulates phosphorylation of Akt at ser473 and phospho ser380 of PTEN, but phosphorylation at ser9 of GSK3β was inhibited. Further, constitutively active (myristoylated) CA-Akt transfected in MRPE cells had an increased Akt activity in EGCG influenced cells. These findings strongly suggest that antioxidant molecules inhibit cell death through the proper balancing of [Ca2+]i and ROS production in order to maintain UPR of ER in MRPE cells. Thus, modulation of UPR signalling may provide a potential target for the therapeutic approaches of AMD.

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Acknowledgments

This study was supported in part by a Grant-in-Aid from Department of Science and Technology (SERB) of India (SB/FT/LS-204/2012) to T. K. B. K. is the recipient of senior research fellowship (09/1012 (0005) 2K11—EMR—I) from the Council of Scientific and Industrial Research (CSIR), India.

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  1. Bose Karthikeyan

    Present address: Department of Ophthalmic Research, Cole Eye Institute, Cleveland Clinic, Cleveland, OH, 44195, USA

Authors and Affiliations

  1. Department of Biotechnology, Kalasalingam University, Anand Nagar, Krishnankoil, Tamil Nadu, 626 126, India

    Bose Karthikeyan, Lakshminarasimhan Harini, Krishnan Sundar & Thandavarayan Kathiresan

  2. Department of Microbiology, Bharathidasan University, Tiruchirappalli, Tamil Nadu, 620 024, India

    Vaithilingam Krishnakumar & Velu Rajesh Kannan

  3. International Research Centre, Kalasalingam University, Krishnankoil, Tamil Nadu, 626 126, India

    Krishnan Sundar & Thandavarayan Kathiresan

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Karthikeyan, B., Harini, L., Krishnakumar, V. et al. Insights on the involvement of (–)-epigallocatechin gallate in ER stress-mediated apoptosis in age-related macular degeneration. Apoptosis 22, 72–85 (2017). https://doi.org/10.1007/s10495-016-1318-2

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