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Reactive Oxygen Species and p38MAPK Have a Role in the Smad2 Linker Region Phosphorylation Induced by TGF-β | ||
| Iranian Journal of Medical Sciences | ||
| مقاله 8، دوره 43، شماره 4، مهر 2018، صفحه 401-408 اصل مقاله (760.86 K) | ||
| نوع مقاله: Original Article(s) | ||
| شناسه دیجیتال (DOI): 10.30476/ijms.2018.40557 | ||
| نویسندگان | ||
| Reyhaneh Niayesh Mehr1؛ Alireza Kheirollah2؛ Faezeh Seif1؛ Parisa Dayati1؛ Hossein Babaahmadi-Rezaei* 1 | ||
| 1Atherosclerosis Research Center, Department of Clinical Biochemistry, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran | ||
| 2Cellular and Molecular Research Center, Department of Clinical Biochemistry, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran | ||
| چکیده | ||
| Background: Transforming growth factor-β (TGF-β) in addition to the C-terminal region can phosphorylate receptor-regulated Smads (R-Smads) in their linker region. The aim of the present study was to evaluate the role of signaling mediators such as NAD(P)H oxidases (reactive oxygen species [ROS] generators), ROS, and ROS-sensitive p38 mitogen-activated protein kinase (p38MAPK) in this signaling pathway in cultured human vascular smooth muscle cells (VSMCs).Methods: The present in vitro study was performed on human VSMCs. Proteins were detected by western blotting utilizing an anti-phospho-Smad2 (Ser245/250/255) rabbit polyclonal antibody and a horseradish peroxidase-labeled secondary antibody. Glyceraldehyde-3-phosphate dehydrogenase was used as a loading control. The phospho-Smad2 linker region (pSmad2L) was detected in all the experimental groups: a control group (untreated group), a group treated with TGF-β (2 ng/mL), and a group treated with TGF-β plus different inhibitors. The data were normalized and presented as mean ± SEM. The statistical analyses were performed using SPSS, version 16.0, and the nonparametric Kruskal–Wallis test. A P value smaller than 0.05 was considered statistically significant.Results: The VSMCs treated with TGF-β (2 ng/mL) showed a time-dependent increase in the pSmad2L level. The highest level was observed at 15 minutes (P=0.03). The inhibitors of NAD(P) H oxidases (diphenyleneiodonium and apocynin) (P=0.04), ROS scavenger (N-acetylcysteine) (P=0.04), and p38MAPK inhibitor (SB-202190) (P=0.04) were able to reduce the increased level of the pSmad2L by TGF-β.Conclusion: Our results suggested that NAD(P)H oxidases played an important role in the Smad2L phosphorylation in the human VSMCs. Furthermore, our results confirmed that ROS and p38MAPK were involved in this signaling pathway. Thus, TGF-β via a ROS-dependent mechanism can transmit its signals to the pSmad2L. | ||
| کلیدواژه‌ها | ||
| Transforming growth factor beta؛ Smad2 protein؛ Reactive Oxygen Species؛ NADPH oxidase 4؛ P38 mitogen-activated protein kinases | ||
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