Reactive Oxygen Species and p38MAPK Have a Role in the Smad2 Linker Region Phosphorylation Induced by TGF-β

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Niayesh Mehr Reyhaneh; KHEIROLLAH ALIREZA; Seif Faezeh; Dayati Parisa; BABAAHMADI REZAEI HOSSEIN

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Journal: IRANIAN JOURNAL OF MEDICAL SCIENCES (IJMS) Year:2018 | Volume:43 | Issue:4 Page(s): 401-408

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Title

Reactive Oxygen Species and p38MAPK Have a Role in the Smad2 Linker Region Phosphorylation Induced by TGF-β

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Keywords

Transforming growth factor beta

Smad2 protein

Reactive oxygen species

NADPH oxidase 4

P38 mitogenactivated protein kinases

Abstract

Background: Transforming growth factor-β (TGF-β ) in addition to the C-terminal region can phosphorylate receptorregulated 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.

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APA: Copy

Niayesh Mehr, Reyhaneh, KHEIROLLAH, ALIREZA, Seif, Faezeh, Dayati, Parisa, & BABAAHMADI REZAEI, HOSSEIN. (2018). Reactive Oxygen Species and p38MAPK Have a Role in the Smad2 Linker Region Phosphorylation Induced by TGF-β. IRANIAN JOURNAL OF MEDICAL SCIENCES (IJMS), 43(4), 401-408. SID. https://sid.ir/paper/758458/en

Vancouver: Copy

Niayesh Mehr Reyhaneh, KHEIROLLAH ALIREZA, Seif Faezeh, Dayati Parisa, BABAAHMADI REZAEI HOSSEIN. Reactive Oxygen Species and p38MAPK Have a Role in the Smad2 Linker Region Phosphorylation Induced by TGF-β. IRANIAN JOURNAL OF MEDICAL SCIENCES (IJMS)[Internet]. 2018;43(4):401-408. Available from: https://sid.ir/paper/758458/en

IEEE: Copy

Reyhaneh Niayesh Mehr, ALIREZA KHEIROLLAH, Faezeh Seif, Parisa Dayati, and HOSSEIN BABAAHMADI REZAEI, “Reactive Oxygen Species and p38MAPK Have a Role in the Smad2 Linker Region Phosphorylation Induced by TGF-β,” IRANIAN JOURNAL OF MEDICAL SCIENCES (IJMS), vol. 43, no. 4, pp. 401–408, 2018, [Online]. Available: https://sid.ir/paper/758458/en

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