Data demonstrating the role of peroxiredoxin 2 as important anti-oxidant system in lung homeostasis

The data presented in this article are related to the research paper entitled “peroxiredoxin-2 plays a pivotal role as multimodal cytoprotector in the early phase of pulmonary hypertension” (Federti et al., 2017) [1]. Data show that the absence of peroxiredoxin-2 (Prx2) is associated with increased lung oxidation and pulmonary vascular endothelial dysfunction. Prx2−/− mice displayed activation of the redox-sensitive transcriptional factors, NF-kB and Nrf2, and increased expression of cytoprotective system such as heme-oxygenase-1 (HO-1). We also noted increased expression of both markers of vascular activation and extracellular matrix remodeling. The administration of the recombinant fusion protein PEP Prx2 reduced the activation of NF-kB and Nrf2 and was paralleled by a decrease in HO-1 and in vascular endothelial abnormal activation. Prolonged hypoxia was used to trigger pulmonary artery hypertension (PAH). Prx2−/− precociously developed PAH compared to wildtype animals.


a b s t r a c t
The data presented in this article are related to the research paper entitled "peroxiredoxin-2 plays a pivotal role as multimodal cytoprotector in the early phase of pulmonary hypertension"   [1]. Data show that the absence of peroxiredoxin-2 (Prx2) is associated with increased lung oxidation and pulmonary vascular endothelial dysfunction. Prx2 −/− mice displayed activation of the redox-sensitive transcriptional factors, NF-kB and Nrf2, and increased expression of cytoprotective system such as heme-oxygenase-1 (HO-1). We also noted increased expression of both markers of vascular activation and extracellular matrix remodeling. The administration of the recombinant fusion Contents lists available at ScienceDirect journal homepage: www.elsevier.com/locate/dib protein PEP Prx2 reduced the activation of NF-kB and Nrf2 and was paralleled by a decrease in HO-1 and in vascular endothelial abnormal activation. Prolonged hypoxia was used to trigger pulmonary artery hypertension (PAH PEP Prx2 significantly reduces protein oxidation in lung from exposed to prolonged hypoxia used to trigger pulmonary artery hypertension.

Data
Data show increased lung oxidation (Fig. 1A) and abnormal pulmonary vascular leakage in the absence of Prx2 (Fig. 1B). This was paralleled by the activation of redox-sensitive transcriptional factors NF-kB and Nrf2 in lung from Prx2 −/− compared to wildtype animals ( Fig. 2A). Indeed, in Prx2 −/ − we observed (i) increased expression of heme-oxygenase 1 (HO-1), a Nrf2 related cytoprotective system; (ii) markers of vascular endothelial activation such as endothelin-1 (ET-1) and vascular cell adhesion molecule -1 (VCAM-1) and (iii) marker of extracellular matrix remodeling as the platelet growth factor-B (PDGF-B) that has been recently function linked to the development of pulmonary artery hypertension (Fig. 2B). To verify the role of Prx2 as important anti-oxidant system in pulmonary homeostasis, we administrated the recombinant fusion protein PEP Prx2 at the dosage of 3 mg/Kg/d ip or vehicle for 4 weeks [1][2][3]. As shown in Fig. 2, PEP Prx2 significantly reduced both NF-kB and Nrf2 activation in lung from Prx2 −/− and decreased the expression of both HO-1 and markers of vascular endothelial activation or extracellular matrix remodeling.
Using prolonged hypoxia to trigger pulmonary artery hypertension, we observed severe lung pathologic damage and the precocious development of pulmonary artery hypertension in Prx2 −/− mice compared to wildtype animals [4][5][6]. This was associated with (i) marked activation of redoxrelated transcriptional factors; (ii) severe endoplasmic-reticulum stress with activation of the unfolded protein response (UPR) system; and (iii) activation of autophagy [1].
PEP Prx2 treatment prevented the hypoxia induced protein oxidation in mice exposed to prolonged hypoxia (7 days; Fig. 3A) and reduced the hypoxia induced increased expression of HO-1 in both mouse strains exposed to 3 days hypoxia (Fig. 3B). Collectively, these data indicate the important role of Prx2 in lung homeostasis against hypoxia, a known trigger of lung injury.

Bronchoalveolar lavage assay
Bronchoalveolar lavage (BAL) fluids were collected and cellular contents were recovered by centrifugation and counted by microcytometry as previously reported [6,9].

Measurement of lung protein oxidation
Oxidized proteins were revealed by the Oxyblot Protein Oxidation Detection Kit (EMD Millipore). In brief, the soluble protein extracts were derivatized to 2,4-dinitrophenylhydrazone (DNP) and 1 µg was loaded on 12% SDS-PAGE, blotted and incubated with an anti-DNP antibody, followed by an HRP conjugated secondary antibody. The bound activity was revealed by ECL (GE Healthcare). Oxidized proteins were revealed by the Oxyblot Protein Oxidation Detection Kit (EMD Millipore). In brief, the soluble protein extracts were derivatized to 2,4-dinitrophenylhydrazone (DNP) and 1 ug was loaded on 12% SDS-PAGE, blotted and incubated with an anti-DNP antibody, followed by an HRP conjugated secondary antibody. The bound activity was revealed by ECL (GE Healthcare) [12][13][14].

Transparency document. Supplementary material
Transparency data associated with this article can be found in the online version at http://dx.doi. org/10.1016/j.dib.2017.09.062.