| Background:Osteoarthritis(OA)is one of the most common diseases faced by joint surgeons and can involve a single joint or multiple joints,especially in the heavily weightbearing knee and hip joints.Symptoms of osteoarthritis include joint pain,stiffness,swelling,and limited mobility,which can often be present for a decade or decades,causing a significant financial burden and seriously affecting the patient’s quality of life.Osteoarthritis is not just a degenerative disease of cartilage but a multifactorial disease affecting the entire joint.In addition to cartilage,the subchondral bone is also an important structure of the knee joint,which adapts to the mechanical stresses acting on the joint by regulating bone remodeling.Academic understanding of the pathogenesis of OA is still evolving,and it is because of the incomplete knowledge of the pathogenesis of OA and the lack of early interventions that the treatment of osteoarthritis is mainly aimed at advanced stages,with arthroplasty being the mainstay.The role of micro RNAs in the pathogenesis of OA has also been extensively studied,and with the discovery of OA-related targets,the role of micro RNAs in the early diagnosis and prevention of OA has become increasingly important.With the discovery of OA-related targets,early diagnosis and prevention of OA are increasingly possible.Objectives:1.To elaborate the differential expression of micro RNAs in the medial and lateral subchondral bone of the tibial plateau in osteoarthritis.2.Predict the target genes of differentially expressed mi RNAs and perform functional enrichment analysis of the target genes.3.To predict the mi RNA/m RNA/key signaling pathways associated with cartilage homeostasis and bone remodeling in subchondral bone.To provide an experimental basis for clarifying the pathogenesis of osteoarthritis and the differences between the medial and lateral compartments of the knee joint in the progression of OA.Methods:1.Screening of differentially expressed mi RNAs: subchondral bone was selected from the tibial plateau of six patients with osteoarthritis who underwent knee arthroplasty,and the subchondral bone on the inner side of the tibial plateau was the experimental group and the outer side was the control group.High-throughput sequencing technology was applied to detect mi RNA expression,and differentially expressed mi RNAs were obtained,and cluster heat map and violin map were drawn.2.Bioinformatics analysis: Target genes of the TOP10 up-regulated and downregulated differentially expressed mi RNAs were predicted using mi RDB(http://www.mirdb.org/)and Mi RWalk(http://mirwalk.umm.uni-heidelberg.de/)databases.GO analysis and KEGG analysis were used to predict the biological processes and signaling pathways involved in the regulation of target genes,and mi RNA-m RNA interaction map and PPI network map were drawn.3.The above analyses were combined to link mi RNAs,predicted target genes and signaling pathways in tandem for discussion and analysis.Results:1.87 differentially expressed mi RNAs were detected by high-throughput sequencing,including hsa-let-7c-3p,hsa-let-7d-3p,hsa-let-7i-3p,hsa-mi R-100-3p,hsa-mi R-101-3p,hsa-mi R-10399-3p,hsa-mi R-122-5p,hsa-mi R-1256,hsa-mi R-1268 b,hsa-mi R-1277-3p,hsa-mi R-1307-5p,hsa-mi R-130a-3p,hsa-mi R-130b-3p,hsa-mi R-142-3p,hsa-mi R-142-5p,has-mi R-144-3p,hsa-mi R-148b-3p,hsa-mi R-150-3p,hsa-mi R-153-3p,hsa-mi R-21-3p,hsa-mi R-210-3p,hsa-mi R-24-1-5p,hsa-mi R-26a-1-3p,hsa-mi R-30e-5p,hsa-mi R-3184-3p,hsa-mi R-32-5p,hsa-mi R-3620-5p,hsami R-374a-5p,hsa-mi R-377-3p,hsa-mi R-3944-3p,hsa-mi R-4466,hsa-mi R-4707-3p,has-mi R-496,hsa-mi R-503-5p,hsa-mi R-548ap-3p,hsa-mi R-548aq-3p,hsa-mi R-548 ba,hsa-mi R-548d-5p,hsa-mi R-570-3p,hsa-mi R-592,hsa-mi R-6715a-3p hsami R-675-3p,hsa-mi R-6824-3p,hsa-mi R-7155-3p,novel-hsa-mi R104-5p,novel-hsami R-158-3p,novel-hsa-mi R199-3p,novel-hsa-mi R223-5p,novel-hsa-mi R243-5p,novel-hsa-mi R26-5p,novel-hsa-mi R269-5p,novel-hsa-mi R288-5p and 52 other mi RNAs were up-regulated in expression.hsa-mi R-10526-3p,hsa-mi R-11401,hsami R-1285-5p,hsa-mi R-1299,hsa-mi R-1323,hsa-mi R-3085-3p,hsa-mi R-34c-3p,hsa-mi R-3529-3p,hsa-mi R-3614-5p,hsa-mi R-3622a-5p,hsa-mi R-3913-5p,hsa-mi R-4524a-3p,hsa-mi R-4634,hsa-mi R-487b-5p,hsa-mi R-512-3p,hsa-mi R-515-3p,hsami R-516a-5p,hsa-mi R-516b-5p,hsa-mi R-517-5p,hsa-mi R-517a-3p,hsa-mi R-517c-3p,hsa-mi R-518 b,hsa-mi R-519d-3p,hsa-mi R-520 h,hsa-mi R-522-3p,hsa-mi R-548ad-5p,hsa-mi R-548 ax,sa-mi R-6761-5p,novel-hsa-mi R131-3p,novel-hsami R176-5p,novel-hsa-mi R215-3p,novel-hsa-mi R294-3p,novel-hsa-mi R35-5p,novel-hsa-mi R53-3p,novel-has-mi R63-5p and 35 other mi RNAs were downregulated.2.Target gene prediction: After the target gene prediction of the top 10 up-and down-regulated mi RNAs by database mi RDB and Mi RWalk and crossed by Venn diagram,a total of 236 genes were predicted to be the target genes of TOP10 up-or down-regulated differentially expressed mi RNAs.3.Bioinformatics analysis: GO analysis identified biological processes(cell morphogenesis involving differentiation,protein phosphorylation,regulation of kinase activity,negative regulation of cell differentiation,enzyme-linked receptor protein signaling pathway)and molecular functions(protein kinase activity,RNA polymerase II specificity,protein kinase binding,DNA-binding transcriptional repressor activity,nuclear receptor activity,transcription.KEGG pathway analysis showed that a total of 119 signaling pathways involved in target genes were significantly altered(P<0.05),some of which were involved in the regulation of cartilage homeostasis,osteogenesis and lipogenesis differentiation.Conclusions:1.A total of 87 mi RNA differential expressions were measured in this study,of which 52 mi RNA expressions were up-regulated and 35 mi RNA expressions were down-regulated.Differential expression of subchondral bone mi RNAs in the medial and lateral tibial plateau of the OA knee was demonstrated.2.A total of 236 m RNAs were predicted to be co-expressed in the two databases.Some of these target genes,such as PPAR-γ,TGF-βRI,TGF-βRII,SMAD2,IGF-II,IGFBP5 and SEMA3 A,are involved in OA related signaling pathways,suggesting that some mi RNAs measured in this study may be involved in OA related signaling pathways by regulating these target genes.3.GO analysis revealed that the biological processes and molecular functions involved in these target genes may be related to adipogenesis,osteoblast differentiation,osteoclast differentiation,and inflammatory response.4.KEGG pathway analysis revealed that Wnt signaling pathway,TGF-β signaling pathway,NF-κB signaling pathway and m TOR signaling pathway were the major enriched signaling pathways of target genes in this study.There were significant associations with osteogenic and lipogenic differentiation,cartilage homeostasis,and bone remodeling.5.Combining the above analysis predicted several mi RNA/m RNA/signaling pathways that could potentially regulate cartilage homeostasis and bone remodeling process: hsa-mi R-130a-3p/PPAR-γ/m TOR signaling pathway,hsa-mi R-130a-3p/TGF-βR1/TGF-βR2/TGF-β signaling pathway,hsa-mi R-210-3p/IGF2/PI3K/Akt signaling pathway,hsa-mi R-210-3p/IGF2/TGF-β signaling pathway,hsa-mi R-3085-3p/IGFBP5/JNK/MEK/Erk signaling pathway,and hsa-mi R-3184-3p/SMAD2/TGF-β signaling pathway.In summary,this study used high-throughput sequencing technology to elucidate the differential expression of mi RNAs in the medial and lateral subchondral bone of the tibial plateau in osteoarthritis,and predicted several potential target genes and signaling pathways regulating cartilage homeostasis and bone remodeling by bioinformatics analysis methods,which have the potential to become molecular targets for early diagnosis of OA,and to further elucidate the mechanism of OA,understand the medial and lateral knee joint These mi RNAs have the potential to become molecular targets for early diagnosis of OA,providing a scientific basis for further elucidating the mechanisms of OA and understanding the differences between the medial and lateral compartments of the knee in OA progression. |
PDF Full Text Request