Eriocalyxin B mediates the migration and inflammation of TGF β2 induced human lens epithelial cells by inhibiting JAK/STAT3 pathway

Abstract

Purpose: To study the role of Eriocalyxin B (EriB) in the migration and inflammation of TGF-β2-induced human lens epithelial cells (hLECs), and to elucidate the molecular mechanisms involved. Methods: The hLECs cultured in vitro were divided into 5 groups, viz, control, TGFβ2, TGFβ2+2 μM EriB, TGFβ2+4 μM EriB, and TGFβ2+8 μM EriB groups. CCK-8, clone formation and Edu labeling assays were performed to assess the effect of EriB on the proliferation of hLECs cells. To determine the role of EriB in cell migration, Transwell and wound healing assays were used. The levels of vimentin, α-SMA, snail, TNF-α，IL-1β，IL-6, P65, p-P65, p-JAK2, JAK2, p-STAT3, STAT3, and β-catenin in hLECs cells were evaluated by enzyme-linked immunosorbent assay (ELISA) and western blot analysis in order ascertain the signaling pathways involved. Results: The rate of cell proliferation significantly decreased in TGFβ2+2μM EriB, TGFβ2+4μM and TGFβ2+8μM groups compared with TGFβ2 group (p < 0.001). In addition, the migration of hLECs cells and epithelial mesenchymal transition were inhibited by EriB in a dose-dependent way (p < 0.001). ELISA results showed that compared to TGFβ2 group, TNF-α, IL-1β and IL-6 levels in EriB group significantly decreased (p < 0.001). The levels of TNF-α, IL-1β, IL-6, p-P65/P65, p-JAK2/JAK2, p-STAT3/STAT3 and metastasis-associated proteins (α-SMA and snail) in hLECs cells were downregulated by EriB (p < 0.001). Furthermore, vimentin level was increased by EriB (p < 0.001). Conclusion: The results show that EriB inhibits the growth, metastasis and inflammation of hLECs cells by inhibiting JAK/STAT3 pathway, thus indicating that this pathway is a potential therapeutic target for treating cataract