Cellar Modified Metabolome Implicated In The Pathogenesis Of Skin-cell Injuries Induced By Different Chemicals

Skin burn has been emerged as a global problem with public health. Generally, many factors daily cause skin burn, e.g.thermal liquid, fire, chemicals, biochemicals, UV/IR radiation, radiation etc. These factors were observed to often cause a multilayer electrochemical or destroy all layers of skin that lead to the formation of scalds. Burns with trauma often induce immune system response syndrome. In patients with severe burn, the plasma extravasation into tissues caused systemically inflammatory response and increased vascular permeability which resulted in tissue edema. Scalds induce metabolic system lesion characterized by increased energy consumption and catabolism, the loss of mark protein, redistributed muscle protein, as well as recombinant organism. Severe burns triggered scar tissue formation and contract across the joints, which will restrict or inactivate joints movement. Then the quality of life with patients were significanty degraded. The molecular mechanism of burns are still pooly understood that markedly impeded the diagnosis, treatment and drug discovery against the burns and associated scalds. In this study, we were first to estabolish the cell injury models of HaCaT induced by different biochemical factors(hydrogen peroxide, phenol and lipopolysaccharide) as an effort was supposed to simulate biochemical processes of the scalds, and then delineated individualized metabolic characteristics of cell injuries by the combinational use of NMR and GC/MS based metabolomics method. The manipulation of this study was supposed to enable us to better understanding of molecular characteristics of the scalds. In addition, we were capable of identifying differential metabolite-biomarkers for monitoring the therapeutic progression and outcomes of the scalds. As well, we have the capacity to verify the rational cell model for further discovery and identification of bioactive compounds derived from Chinese medicinal plants and formulae to treat the scalds at a clinical setting.We developed skin cell models with injury induced by H2O2, PH and LPS that were employed to stimulate the scalds in a real world. NMR and GC/MS based metabolomics method were optimized and developed that allows us to high-efficiency investigate modified metabolome in skin cells. Multiple pattern recognition methods were selected to manipulate group classification and clustering, and recognize differentiable mebolites whose level changes can phenotype the onsite and progression of skin cell injury. Bioinformatic assay was performed to construct metabolic pathways for the differential metabolites, and functional annotation was facilitated to interrogate biological fucntions and significance of modified metabolies in the pathogenesis of skin cell injuries.To annotate metabolic characteristics of skin cell injuries caused by differentiablly chemical factors, the developed NMR and GC/MS based metabolomics method was exploited for characterizing the individualized metabolome that were closely associated with the treatment of injury factors(H2O2, PH and LPS). Our data manifested that 27 metabolites were clearly observed to link to the cell injuries induced by the defined chemicals. These metabolites primarily include amino acids(alanine, asparagine, arginine, glutamine, valine, histidine, proline, leucine, isoleucine, norleucine, ornithine, aspartic acid, and glutamic acid), energetic cycle associated metabolites(malic acid, citric acid, glucose, allose, pyruvate, glycerol, stearic acid, lipid, lactic acid, and furanose) and others(creatine, choline, adenosine, creatinine, and glutathione). Some of above differential metabolites were discovered and identified by the NMR based metabolomics, including histidine, proline, lactic acid, glutathione, lysine, ornithine, asparagine, creatine, citric acid, lipid, aspartic acid, alanine, choline, malic acid, arginine, asparagine, adenosine, valine, and glutamine metabolites. Others were characterized by the GC/MS based metabolomics method involving aspartic acid, glycerol, leucine, lactic acid, glutamic acid, allose, fructose, lysine, glucose, serine, proline, glutamine, alanine, maltose, norleucine, pyruvic acid, stearic acid, glycerol, isoleucine, and valine metabolites. In addition, many metabolites such as proline, alanine aspartic acid, glutamine, valine and lysine were determined together by two metabolomics methods. Furthermore, we homed them to their metabolic pathways, and the results demonstrated that they mostly belong to glycolysis, lipid metabolism, pentose phosphate pathway, citric acid cycle, and urea cycle. This might suggested that skin cell injuries primarily incurred modified metabolism involving energetic metabolism, DNA/RNA biosynthesis, and membrance construction, which is almost agreeable with the biochemical processes of the scalds in clinical context. At last, our data further verified that the cell model induced by the altered levels of H2O2 incurred mostly wide-spectrum metabolic perturbations that were highly consistent with the imaging observation on cell injury, as well as rendered obvious dependency of treatment concentration.Altogether, this study was first to phenotype metabolic charateristics of cell injuries induced by different chemicals that was closely consistent with clinical phenome of the scalds, and verified the in vitro model of cell injury induced by H2O2 as the optimized one that has the capacity to serve for interrogating the pathogenesis, and diagnostic monitoring of the scalds, as well as high-throughput screening of bioactive compounds derived from Chinese medicinal plants and formulae to treat the skin issues.