Metabolomics study of fibroblasts damaged by UVB and BaP

Abstract

We have recently shown that both UVB and BaP can induce the production of ROS, apoptosis and even cancer. However, the differences in the metabolic profiles of skin damaged by UVB, BaP or UVB combined with BaP have not been studied. Therefore, we examined the metabolic changes in the human foreskin fibroblast injured by UVB or BaP or the combination of the two, using ultra performance liquid chromatography (UPLC) coupled with quadrupole time-of-flight mass spectrometry (qTOF-MS). 24 metabolites were altered in the UVB damage group, 25 in the BaP damage group, and 33 in the UVB combined with BaP group. These alterations indicated that the metabolic mechanisms of HFF-1 cells treated with UVB or BaP are related to multiple main metabolites including glycerophosphocholine (PC), lactosylceramide (LacCer), guanidinosuccinic acid (GSA), glutathione(GSH), and lysophosphatidylcholine (LysoPC) and the main mechanisms involved glycerophospholipid and glutathione metabolism. Thus, our report provided useful insight into the underlying mechanisms of UVB and BaP damage to skin cells.

Figure 1

Principal components analysis for metabonomics of damaged cell models. (a) Cytotoxicity of HFF-1 exposed to different UVB irradiation intensities. (b) BaP induced reactive oxygen species production. Cells were exposed to 5–40 mJ/cm² UVB irradiation and the doses of BaP ranged from 10 nM to 100 µM. (c) PCA score plots. (d–f) OPLS-DA score plots based on the data from UPLC-ESI (+)-QTOF-MS for distinguishing C versus U (R2Y = 99.6%, Q2 = 85.6%), C versus B (R2Y = 99.6%, Q2 = 86.1%), and C versus D (R2Y = 99.4%, Q2 = 86.9%) for selecting potential markers.

Figure 2

Differential metabolites of damaged cell models. (a) Overlap analysis of metabolites differing between damaged groups and the control group. (b) Pathway analysis showing altered metabolic pathways. C, control group; U, UVB exposure group; B, BaP exposure group; D, UVB and BaP group.

Figure 3

Boxplot of the altered metabolites. GSA was main biomarker in C versus U and C versus D. LysoPE, LysoPC and GSH were the main biomarkers in C versus B and C versus D. GSSG and the ratio of GSSG/GSH represented oxidative stress of cells, the larger the ratio of GSSG/GSH, the stronger the oxidation. C, control group; U, UVB exposure group; B, BaP exposure group; D, UVB and BaP group.