Leptin activates the JAK/STAT pathway to promote angiogenesis in RF/6A cells in vitro
Authors:   Department of Ophthalmology, Northwest Woman's and Children's Hospital, Xi'an 710061, Shaanxi Province, China; Department of Ophthalmology, Shaanxi Provincial People's Hospital, Xi'an 710068, Shaanxi Province, China, Zhang Le , Wu Bing-Hui ,   Department of Ophthalmology, Xi'an No.1 Hospital, Xi'an 710001, Shaanxi Province, China, Liang Ting-Ting ,   Medical College of Xi'an Medical University, Xi'an 710021, Shaanxi Province, China, Liu Zhe ,   Department of Medical Interdisciplinary Research, Xi'an Ninth Hospital Affiliated to Medical College of Xi'an Jiaotong University, Xi'an 710054, Shaanxi Province, China, Ju Wei ,   Department of Medical Interdisciplinary Research, Xi'an Ninth Hospital Affiliated to Medical College of Xi'an Jiaotong University, Xi'an 710054, Shaanxi Province, China, Wang Yi ,   Department of Medical Interdisciplinary Research, Xi'an Ninth Hospital Affiliated to Medical College of Xi'an Jiaotong University, Xi'an 710054, Shaanxi Province, China, Wen Yu-Ting ,   Department of Medical Interdisciplinary Research, Xi'an Ninth Hospital Affiliated to Medical College of Xi'an Jiaotong University, Xi'an 710054, Shaanxi Province, China, Liu Ming-Cui ,   Department of Medical Interdisciplinary Research, Xi'an Ninth Hospital Affiliated to Medical College of Xi'an Jiaotong University, Xi'an 710054, Shaanxi Province, China, Du Jun-Hui ,   Department of Medical Interdisciplinary Research, Xi'an Ninth Hospital Affiliated to Medical College of Xi'an Jiaotong University, Xi'an 710054, Shaanxi Province, China; Department of Ophthalmology, Xi'an Ninth Hospital Affiliated to Medical College of Xi'an Jiaotong University, Xi'an 710054, Shaanxi Province, China
Published: March 2022

AIM: To investigate the effect of leptin on the angiogenesis of RF/6A cells (monkey retinal choroidal endothelial cells) in vitro and test the cellular signaling in the mechanism.
METHODS: RF/6A cells were cultured in vitro and randomly divided into four groups: normal control, with leptin at 50, 100, 200 ng/mL for cell counting kit-8 (CCK8). RF/6A cell proliferation and migration were examined by Transwell assays, while RF/6A cell tube formation by Matrigel assay. JAK2, p-JAK2, STAT3, and p-STAT3 protein expression was measured by Western blotting. Cells were then divided into the following treatment groups: control, 100 ng/mL leptin and AG-490 (100 ng/mL leptin+10 μmol/L AG-490) for examinations of RF/6A cellular behaviour again. Analysis of differences was carried out using one-way ANOVA and least significant difference (LSD).
RESULTS: RF/6A cell proliferation, migration and cell tube formation were promoted significantly by leptin in a dose-dependent manner (P&#x003C;0.05). Western blotting showed that leptin up-regulated p-JAK2 and p-STAT3 expression levels. Treatment with the JAK/STAT pathway inhibitor, AG-490, decreased leptin-induced p-JAK2 and p-STAT3 expression, and inhibited cell proliferation, migration and cell tube formation induced by leptin (P&#x003C;0.05).
CONCLUSION: Leptin can promote RF/6A cell angiogenesis in vitro via activation of the JAK2/STAT3 signaling pathway.