Study On The Wound-age Estimation Of Skeletal Muscle Early Injury Based On Metabolomics

Objective:Metabolic characteristics were different between skeletal muscle after contusion and normal skeletal muscle in rats.To find out the potential biomarkers for wound-age estimation of early skeletal muscle contusion in rats.To find out the metabolic pathways changed by skeletal muscle early after contusion in rats.Methods:To establish the rat right hindlimb skeletal muscle contusion model.Including skeletal muscle contusion 0.5h group,skeletal muscle contusion 1h group,skeletal muscle contusion 2h group and control group(n=7).Approximately 200 mg muscle was dissected from each injured skeletal muscle at 0,6,12 and 24 h post-mortem.The sample were analyzed by GC/MS.The MS-Dial ver2.76 were used for original spectrum preprocessing.Statistical data analysis was performed using SIMCA-P14.1 combined with OPLS-DA and one-way analysis of variance(ANOVA)to pick out characteristic biomarkers associated with wound age.Results:Metabolic characteristics were different between injuryed skeletal muscle and normal skeletal muscle in rats.Five kinds of potential biomarkers were selected from the experimental group,which were N-FormylGlycine,Malonic acid,Sarcosine,Alanyl-alanine,Cadaverine and showed a time-related expression.9 different metabolites were screened from the contusion 0.5h model group,respectively;8 different metabolites were screened from the contusion 1h model group,respectively;10 different metabolites were screened from the contusion 2h model group,respectively.A set of skeletal muscle contusion 0.5h group muscle biomarkers composed of L-leucine,DOPA,Sarcosine,Cadaverine,N-formylglycine,Malonic acid,L-serine,Glycine acid,Maltoriose was obtained through the ROC curve analysis;A set of skeletal muscle contusion 1h group biomarkers composed of L-leucine,DOPA,Sarcosine,Cadaverine,N-formylglycine,Malonic acid,Aconitic acid,Oxalic acid was obtained through the ROC curve analysis.A set of skeletal muscle contusion 2h group biomarkers composed of L-leucine,DOPA,Sarcosine,Cadaverine,N-formylglycine,Malonic acid,Aconitic acid,D-threitol,serotonin,D-Xylulose was obtained through the ROC curve analysis.The affected mainly metabolic pathways in rats were Tricarboxylic Acid Cycle,glycolysis,pentose phosphate pathway and amino acid metabolism,respectively,and are related to energy metabolism,protein decomposition and synthesis.Conclusions:In this study,the metabolic profile in rats skeletal muscle after contusion were investigated based on metabolomics,and the metabolites from the rats may be the biomarkers for estimation of wound age.The altered metabolic pathway including Tricarboxylic Acid Cycle,glycolysis,pentose phosphate pathway and amino acid metabolism,respectively,and are related to energy metabolism,protein decomposition and synthesis were found in the rats of skeletal muscle after contusion.The results provide a foundation and strategies for the forensic identification of wound-age estimation.