Ritonavir inhibits HIF-1?-mediated VEGF expression in retinal pigment epithelial cells in vitro.

ABSTRACT:

Purpose

Retinal hypoxia-mediated activation of the hypoxia-inducible factor (HIF pathway) leading to angiogenesis is a major signaling mechanism underlying a number of sight-threatening diseases. Inhibiting this signaling mechanism with an already approved therapeutic molecule may have promising anti-angiogenic role with fewer side effects. Hence, the primary objective of this study was to examine the expression of HIF-1? and VEGF in human retinal pigment epithelial cells treated with ritonavir under hypoxic and normoxic conditions.

Methods

ARPE-19 and D407 cells were cultured in normoxic or hypoxic conditions, alone or in the presence of ritonavir. Quantitative real-time polymerase chain reaction, immunoblot analysis, sandwich ELISA, endothelial cell proliferation, and cytotoxicity were performed.

Results

A 12-h hypoxic exposure resulted in elevated mRNA expression levels of both HIF-1? and VEGF in ARPE-19 and D407 cells. Hence, this time point was selected for subsequent experiments. Presence of ritonavir in the culture medium strongly inhibited VEGF expression in a concentration-dependent manner under hypoxic conditions. Immunoblot analysis demonstrated a substantially reduced protein expression of HIF-1? in the presence of ritonavir. Further, hypoxic exposure-induced VEGF secretion was also inhibited by ritonavir, as demonstrated using ELISA. Finally, ritonavir significantly diminished the proliferation of choroid-retinal endothelial (RF/6A) cells demonstrating potential anti-angiogenic activity. Cytotoxicity studies showed that ritonavir is non-toxic to RPE cells.

Conclusions

This study demonstrates for the first time that ritonavir can inhibit HIF-1? and VEGF in ARPE-19 and D407 cells. Such inhibition may form a platform for application of ritonavir in the treatment of various ocular diseases.

SUBMITTER: Vadlapatla RK 

PROVIDER: S-EPMC3890766 | biostudies-literature | 2014 Jan

REPOSITORIES: biostudies-literature