Expression Of Noncoding RNA And Bioinformatics Analysis In Tibialis Anterior Muscle Wasting At The Early Stage After Severe Scald Rats Performed By Microarray

Background:Since twenty-first Century, epigenetic research has been paid more and more attention in medicine. With the development of bioinformatics technology, the etiology, diagnosis, of disease of biochip technology, has become a new research method, applied in clinical and basic research more widely. Our previous study found that:after severe burn high metabolic consumption existed in skeletal muscle, and the tibialis anterior(TA) muscle consumption is more significant, further study found that the ubiquitin proteasome mediated protein degradation, apoptosis of skeletal muscle cell, endoplasmic reticulum stress are the main reason that skeletal muscle wasting in burns. But slightly less, the epigenetic mechanisms of skeletal muscle wasting in burns have not been studied.Objective:The high-throughput microarray screening was performed at the early stage of severe burn in rat tibialis anterior muscle of non coding RNA expression profile changes, application of bioinformatics, dig the chip data provide information, find the possible pathogenic mechanism, provide new targets and pathways for clinical treatment.In this study, three parts in the following:(1) To observe the early stages of severe scald rat tibialis anterior muscle consumption changes;(2) Screening of severely scalded rats at the early stage of tibialis anterior muscle of miRNAs expressions and bioinformatics analysis;(3) Screening lncRNAs expressions and bioinformatics analysis.early stage of tibialis anterior muscle of severe burn in rat.Methods:(1) In male Wistar rats using the preparation of scalded rats model, with94℃hot water,12s caused rats back30%TBS A scald third degree, the control group rats using37℃warm water (the other conditions with experimental group). After injury, Id,3d,7d,14d (n=8) rats were sacrificed, isolation of tibialis anterior muscle weight, weighing after anesthesia. TUNEL for the determination of the tibialis anterior muscle cell apoptosis. At the same time, preserve the tibialis anterior muscle specimens for detection of miRNAs and lncRNAs.(2) The Danish lock Exiqon miRCURY TM LNA microRNA oligonucleotide chip was performed(the seventh generation), about700capture probes, databases was version V18.0, detection of tibialis anterior in the experimental group and the control group take the injured after3days.(n=4).Selecting some of the changes in microRNAs target gene prediction, cluster analysis, functional analysis of genes,signal pathway analysis and bioinformatics analysis.(3) Using rat long noncoding RNA chip designed by Arraystar Rat LncRNA Array (4x44K, Arraystar),including LncRNAs and protein coding genes,about15000mRNA or9000lncRNAs from UCSC database and NCBI RefSeq database. Detection tibialis anterior muscles in the experimental group and the control group after3days of burn (n=3), gene function analysis on the difference of selected LncRNAs, pathway analysis and deep bioinformatics analysis.Results:(1) Compared with control group at the same time, tibialis anterior muscle weight and the weight ratio declined, at3d of severely burned rats, decline to the lowest at7d; reflecting obvious skeletal muscle wasting state. TUNEL indicated that the tibialis anterior in experimental group showed partly nuclear changes of apoptosis. After injury in tibialis anterior muscle cell apoptosis increased.(2) Compared the scald group and control group, there were69miRNAs changes in two group, which elevated were4miRNAs,while reduced were65miRNAs.Through the GO analysis showed,6miRNAs involved in metabolism,5miRNAs related to the biological regulation. PATHWAY analysis showed that the pathways is closely related to P53signaling pathway, apoptosis signal pathway, PI3K/Akt signaling pathway,and so on.(3) Though the long noncoding RNA microarray data, detecting the differences between the experimental group and the control group of mRNAs from about15000 mRNAs in rats, the experimental group compared with the control group increased significantly, a total of93mRNAs, while the lower is110mRNAs. Detection of differences between the experimental group and the control group of lncRNAs from about9000lncRNAs in rats, the experimental group compared with the control group, increased a total of64lncRNAs, while the lower is53lncRNAs, and the ratio is greater than1.5(P<0.05). GO analysis showed that the negative regulation of biological process, the complement activation classical pathway, voltage-gated chloride channel activity and other functions related to lncRNA. PATHWAY analysis showed that the insulin signaling pathway, T cell receptor signaling pathway may be regulated by lncRNAs.Conclusion:(1) After severe scalding in rats early anterior tibial muscle miRNAs expression profile changes, and bioinformatics analysis showed that miRNAs may mainly involved in the regulation of metabolism process, biological regulation and other functions, and participate in the P53signaling pathway, apoptosis signaling pathways, PI3K/Akt signaling pathway and other signaling pathways.(2) In tibialis anterior muscle early after severe scalded rats, lncRNAs and mRNA changes of expression, and bioinformatics analysis showed that lncRNAs may mainly involved in the biological process of negative regulation, the classic way of complement activation, voltage-gated chloride ion channel activity and other functions, and also participate in the insulin signaling pathway, T cell receptors signaling pathways and other signaling pathways.