title: PI3K-mediated glioprotective effect of epidermal growth factor under oxidative stress conditions.
abstract: OBJECTIVE
  To determine the effects of epidermal growth factor (EGF) on the proliferation and migration of Müller cell line Moorfields/Institute of Ophthalmology-Müller 1 (MIO-M1), and its related molecular mechanisms under normal and oxidative stress conditions.
  
  
  METHODS
  Müller cells were cultured with different concentrations of EGF in the presence or absence of varied amounts of H2O2 and glucose oxidase (GO) which induced oxidative stress. The proliferation and migration of Müller cells were examined by 5-Bromo-2-deoxyUridine (BrdU), MTT assay, Transwell assay and scratch wound healing assays. The cell viability was determined with the MTT assay. The secretion of EGF by Müller cells was evaluated by ELISA. Western blot was performed to detect the activation of extracellular regulated protein kinases (ERK)1/2 and Akt signal pathways.
  
  
  RESULTS
  EGF stimulated the proliferation and migration of Müller cells in a concentration-dependent manner in vitro. Under oxidative damage condition, 2h of pretreatment with 10-100 ng/mL EGF can mostly inhibit 50% lethal dose of 0.08 mmol/L H2O2-induced cell damage. The Western blot results showed that after Müller cells were exposed to varying EGF for 24h, Akt and ERK1/2 were phosphorylated in a dose-dependent manner. In the presence of the LY294002, the potent PI3K inhibitor, the p-Akt was significantly attenuated.
  
  
  CONCLUSIONS
  EGF may induce the proliferation and migration of human Müller cells through the Akt and the ERK1/2 signal pathways, and induce PI3K-mediated glioprotective effect under oxidative stress.
authors: School of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical College, Wenzhou 325027, Zhejiang Province, China.
authors: Zhi-Xiang
authors: ZX
authors: Hu
authors: School of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical College, Wenzhou 325027, Zhejiang Province, China.
authors: Chun-Li
authors: CL
authors: Chen
authors: School of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical College, Wenzhou 325027, Zhejiang Province, China.
authors: Jia-Song
authors: JS
authors: Yang
authors: School of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical College, Wenzhou 325027, Zhejiang Province, China.
authors: Zhong-Lou
authors: ZL
authors: Zhou
authors: School of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical College, Wenzhou 325027, Zhejiang Province, China.
authors: Zong-Ming
authors: ZM
authors: Song
authors: Department of Ophthalmology, Zhejiang University School of Medicine, Hangzhou 310058, Zhejiang Province, China.
authors: Zhao-Yang
authors: ZY
authors: Wang
keywords: PI3K; epidermal growth factor; human Müller cells