Simvastatin and amfenac act as immunomodulators during induced epithelial‐mesenchymal transition in ARPE‐19 cells

Abstract

Purpose: Rhegmatogenous retinal detachment can develop into proliferative vitreoretinopathy (PVR) after surgery. Inflammation contributes to epithelial‐mesenchymal transition (EMT) in retinal pigment epithelial (RPE) cells, which takes place in PVR. The aim here was to explore the effects of simvastatin and amfenac on inflammation during the EMT.Methods: EMT in ARPE‐19 cells was stimulated by the commercial Stem X Vivo EMT supplement (R&D Systems) either in non‐inflammatory or inflammatory conditions induced by 2.5 pg/mL IL‐1β (Gibco) or 10 pg/mL IL‐1α (R&D Systems). Cells were treated concurrently with 5 μM simvastatin (Sigma Aldrich), 0.1 μM amfenac (Cayman Chemical) or both. The release of interleukin (IL)‐6 and IL‐8 (BD Biosciences), as well as monocytic chemoattractant protein‐1 (MCP‐1; Thermo Fisher Scientific) were measured using commercial enzyme‐linked immunosorbent assays. Lactate dehydrogenase (LDH) release and intracellular ATP levels were determined using commercial kits (Promega). Cells were photographed under an Axio Vert A1 Zeiss microscope.Results: Simvastatin alone or with amfenac reduced the release of IL‐6 in inflammatory and non‐inflammatory conditions upon experimental EMT, increased the IL‐8 release upon IL‐1α exposure and MCP‐1 in non‐inflammatory conditions and upon IL‐1β exposure. During EMT process, LDH release was increased by simvastatin alone and together with amfenac. They also increased ATP levels without inflammation and with IL‐1α. Amfenac had no effects on the cytokine levels (IL‐6, IL‐8, MCP‐1), LDH release, or ATP upon EMT process without inflammation. However, amfenac increased IL‐6 and MCP‐1 release and reduced LDH levels upon IL‐1β, and reduced IL‐6 upon the IL‐1a exposure. Under microscope, cells appeared alive and bigger but partly detached upon simvastatin.Conclusions: Simvastatin and amfenac showed immunomodulatory effects on ARPE‐19 cells during EMT. Further studies are needed concerning the effects of drugs on the EMT.