Correlation Analysis Of Key MiRNA And MRNA In The Differentiation Of Mongolian Horse BMSCs Into Chondrocytes

Bone Marrow Mesenchymal Stem Cells(BMSCs)are a kind of pluripotent adult stem cells.They exist in the bone marrow matrix and can differentiate into bone cells,chondrocytes,muscle cells,adipocytes,cardiomyocytes,etc.BMSCs have become into the ideal seed cells in tissue engineering and regenerative medicine because of their abundant content and easy separation.There is almost no nerve distribution and blood energy supply in mammalian articular cartilage,so cartilage regeneration ability is poor,and it is difficult to repair itself when injured.Therefore,BMSCs have important value in the treatment of cartilage injuries caused by exercise in mammals.MicroRNA(miRNA)is a class of non-coding RNAs with regulatory function,and its length is about 20-25 nt.Studies have shown that miRNA can silence or reduce the expression of target genes,thus affecting their function.At present,the regulation mechanism of miRNA mediated in the process of BMSCs differentiation into chondrocytes is mainly concentrated in human,mice and rats.There is no systematic research and report on miRNA in horse BMSCs in the process of chondrogenic differentiation.Therefore,in this study,Mongolian horse BMSCs differentiated into chondrocytes at different stages(0d,14d,21d)were took as the research object.Through transcriptome sequencing and bioinformatics analysis,the differential mRNA,miRNA and target mRNA related to the differentiation process were screened out,and their regulatory mechanisms were explored.The main results are as follows:1.From the sequence data of induced differentiation from 0d-14d,0d-21d and 14d-21d,4011,3592 and 1049 differentially expressed mRNAs were screened,and the differentially expressed miRNAs were 1119,1158 and 133,respectively.2.The target genes of 134 miRNAs were predicted,and six key miRNAs,including eca-miR-155_R+1、eca-miR-221、eca-miR-222_R+3,eca-miR-16,eca-miR-27b and eca-miR-148b-3p,were screened by enrichment analysis of GO and KEGG.Four key miRNA-mRNA combinations,ZBTB16(BOC)-eca-miR-155_R+1-PIK3R1-eca-miR-221(eca-miR-222_R+3),eca-miR-16-NFATC2,eca-miR-27b-BHLHE41 and eca-miR-148b-3p-TGFA were obtained,and mRNA-miRNA association network diagram was drawn by interaction analysis of miRNA and its target gene.3.PIK3R1 was the target of eca-miR-155_R+1,eca-miR-221 and eca-miR-222_R+3,and it was predicted that it could inhibit chondrocyte apoptosis and matrix catabolism by PI3K/AKT signaling pathway,furthermore could regulate chondrogenic differentiation of BMSCs.The above results can provide theoretical support for elucidating the mechanism of differentiation of Mongolian horse BMSCs into chondrocytes,and provide theoretical basis and technical methods for horse BMSCs in clinical cell therapy of sports injury,meanwhile provide reference for cell therapy of human clinical medicine.