Characterization of TGF-β by Induced Oxidative Stress in Human Trabecular Meshwork Cells

Antioxidants

Mei-Lan Ko¹  Hsiang-Yu Kung¹  Jia-Yu Chen¹  Xin-Ru Yu¹  Hsiu-Chuan Chou¹  Shing-Jyh Chang²  Yi-Jung Ho³  Yu-Shan Wei⁴  Chu-Chun Yang⁴  Hsin-Yi Chen⁴  Meng-Wei Lin⁴  En-Chi Liao⁴  Yi-Shiuan Wang⁴  Hong-Lin Chan⁴  Yu-An Chien⁴ 

[1] Department of Biomedical Engineering and Environmental Sciences, National Tsing Hua University, Hsinchu 300, Taiwan;Department of Obstetrics and Gynecology, Hsinchu MacKay Memorial Hospital, Hsinchu 300, Taiwan;Department of Ophthalmology, National Taiwan University Hospital Hsin-Chu Branch, Hsinchu 300, Taiwan;Institute of Bioinformatics and Structural Biology & Department of Medical Sciences, National Tsing Hua University, Hsinchu 300, Taiwan;

关键词: TGF-β signal pathway;    oxidative stress;    trabecular meshwork cells;    fibrosis;   

DOI  :  10.3390/antiox10010107

来源: DOAJ

【 摘 要 】

Oxidative stress generated by reactive oxygen species (ROS) plays a critical role in the pathomechanism of glaucoma, which is a multifactorial blinding disease that may cause irreversible damage within human trabecular meshwork cells (HTMCs). It is known that the transforming growth factor-β (TGF-β) signaling pathway is an important component of oxidative stress-induced damage related to extracellular matrix (ECM) fibrosis and activates cell antioxidative mechanisms. To elucidate the dual potential roles and regulatory mechanisms of TGF-β in effects on HTMCs, we established an in vitro oxidative model using hydrogen peroxide (H₂O₂) and further focused on TGF-β-related oxidative stress pathways and the related signal transduction. Via a series of cell functional qualitative analyses to detect related protein level alterations and cell fibrosis status, we illustrated the role of TGF-β1 and TGF-β2 in oxidative stress-induced injury by shTGF-β1 and shTGF-β2 knockdown or added recombinant human TGF-β1 protein (rhTGF-β1). The results of protein level showed that p38 MAPK, TGF-β, and its related SMAD family were activated after H₂O₂ stimulation. Cell functional assays showed that HTMCs with H₂O₂ exposure duration had a more irregular actin architecture compared to normal TM cells. Data with rhTGF-β1 (1 ng/mL) pretreatment reduced the cell apoptosis rate and amount of reactive oxygen species (ROS), while it also enhanced survival. Furthermore, TGF-β1 and TGF-β2 in terms of antioxidant signaling were related to the activation of collagen I and laminin, which are fibrosis-response proteins. Succinctly, our study demonstrated that low concentrations of TGF-β1 (1 ng/mL) preserves HTMCs from free radical-mediated injury by p-p38 MAPK level and p-AKT signaling balance, presenting a signaling transduction mechanism of TGF-β1 in HTMC oxidative stress-related therapies.

【 授权许可】

Unknown