The Effect Of ARE Structures On MRNA Expression Heterogeneity After Skeletal Muscle Contusion

Objective:To explore the difference of expression heterogeneity between the AREs-mRNAs and non-AREs mRNAs and provide reference for screening indicators of wound age estimation.Methods:All 108 Spraguee Dawley male rats were separated to control group and post-contusion 4h,8h,12 h,16h,20 h,24h,28 h,32h,36 h,40h,44 h and 48 h groups.This study contained 3 teams,experimental team(n=6),random sampling team(n=3)and add sample team(n=6+2).Random sampling team was obtained with random sampling 3 samples from each groups of experimental team by INDIRECT function of Excel;add sample team added 6 samples to control group and 2 samples to each post-contusion groups based on experimental team.On the basis of sequencing data,AREs-mRNAs and non-AREs mRNAs were screened for 5 mRNAs respectively,RT-qPCR was used to proof the expression level of mRNAs.Subsequently,to measure the heterogeneity difference between the AREs-mRNAs and non-AREs mRNAs,the CV,DFA,STD and Z.SCORE of mRNAs were calculated and used for Fisher discrimination analysis,Random Forest classification and T test.And to compare the heterogeneity size of AREs-mRNAs and non-AREs mRNAs,the Mean CV'CV and CV scores were figured.CV'CV was the variable coefficient of the 13 CV,and Mean CV'CV was the CV'CV mean of AREs-mRNAs and non-AREs mRNAs.Results:For experimental team,both Fisher discrimination analyses,random Forest classification of CV,DFA and STD could recognize AREs-mRNAs from non-AREs mRNAs;T test verified the differences of expression heterogeneity between the AREsmRNAs and non-AREs mRNAs.Mean CV'CV of non-AREs m RNAs is lower than AREs-mRNAs' Mean CV'CV.For random sampling team,the difference of non-AREs mRNAs and AREs-mRNAs is unapparent than experimental team,and its CV'CV and Mean CV'CV were greater than experimental team.With add of samples,discrimination analyses and random Forest classification still could distinguish AREs-mRNAs and non-AREs mRNAs precisely,and the heterogeneity difference between AREs-mRNAs and non-AREs mRNAs was still significantly.Mean CV'CV of non-AREs mRNAs was less than AREs-mRNAs.Conclusion:There was significantly difference between the heterogeneity of AREs-mRNAs and non-AREs mRNAs,and the expression heterogeneity of non-AREs m RNAs was lower than AREs-mRNAs,made non-AREs mRNAs are more suitable for rats skeletal muscle wound age estimation.