Study On The Expression Of Long Noncoding RNAs And Circular RNAs Of Osteoarthritis And Assessment Of Treatment Efficacy Of A Combination Of Glucosamine And Atorvastatin On Osteoarthritis

Part 1:Study on the expression of long noncoding coding RNAs and circular RNAs of osteoarthritis.Background:Long noncoding RNAs?lncRNAs?and circular RNAs?circRNAs?are considered a novel class of regulatory factors in many diseases,but their biological function and the signaling pathway involved in osteoarthritis?OA?remain unknown.Objective:To identify the lncRNAs and circRNAs differentially expressed in OA cartilage,predict the regulatory network and predict the interactions.Methods:The expression of lncRNAs and circRNAs in OA cartilage was measured using an illumina sequencing platform.Furthermore,differentially expressed transcripts of uncertain coding potential?TUCPs?and messenger RNAs?mRNAs?were identified.The colocated target genes and the possible role of lncRNAs as a competing endogenous RNA?ceRNA?were predicted.Protein interaction analysis and ceRNA regulatory network analysis were performed to identify lncRNAs associated with cartilage metabolism of OA.qRT-PCR was done for verification.Differentially expressed circRNAs were identified,and the target miRNA,the source gene,and the circRNA-miRNA-mRNA pair were predicted.Results:Based on 38 cartilage samples from 19 osteoarthritis patients of knees?median age 64.5 years,range 58-75 years,2 males,17 females?,580 significantly dysregulated lncRNAs as well as differently expressed TUCPs and mRNAs were identified.Four differently expressed lncRNAs?SNHG5,ZFAS1,GAS5,and DANCR?involved in OA cartilage were analyzed by protein-protein interactions network and ceRNA regulatory network.Part of our results were consistent with previous studies,and 96 novel differently expressed lncRNAs were identified in OA.197 differentially expressed circRNAs such as hg38 circ 0007474 and hg38_circ₀000118 were identified,and 21 target miRNAs,2466 source genes and 166,394 circRNA-miRNA-mRNA pairs were predicted.Further analysis was applied on three OA related circRNAs?hsa_circ₀045714,hsa_circ₀002485 and hsa_circ₀005567?.The results were partly consistent with previous studies.Conclusion:Our results of the lncRNAs and circRNAs profiling of OA could expand our knowledge of the OA mechanism.Further biological function research are needed to unfold the possible pathway and therapeutic target of OA.Part 2:Assessment of treatment efficacy of a combination of glucosamine and atorvastatin on osteoarthritis.Background:Atorvastatin,a cholesterol-lowering medicine,has been suggested to have a potential therapeutic role in treating osteoarthritis?OA?due to its alti-inflammatory effect.Glucosamine is widely used in the clinical practice as cartilage supplementation in preventing OA progression.However,up to now,the conservative treatment for OA is still unsatisfactory.Objective:Atorvastatin and glucosamine possibly have a synergistic effect in treating progressing OA in a combined administration.To test this hypothesis,we investigated the effectiveness of atorvastatin and glucosamine treatments,respectively,and a combination of them on OA model,both in vitro and in vivo.Methods:We evaluated the regulation of matrix metalloproteinase?MMP?-3,13 and the expression of aggrecan and type ? collagen?COL2?and Aggrecan?AGC?synthesis by Atorvastatin and/or glucosamine in the presence of IL-1? using cultured chondrocytes obtained from human articular cartilage.Then,the in vivo study was performed on the monosodium iodoacetate?MIA?-induced osteoarthritic Sprague Dawley rat.After administration of 8 weeks of Atorvastatin and/or glucosamine,OA severity of the knees by Safranin O-Fast Green staining and TRAP staining were determined.Results:Atorvastatin and glucosamine can reduce MMP-3 expression levels in cultured human chondrocytes,respectively.The combination of the two can further reduce the expression level of MMP-3.The combination of atorvastatin and glucosamine can increase the AGC level expressed by human chondrocytes in vitro.In the MIA-induced rat osteoarthritis model,the combined administration of atorvastatin and glucosamine for 8 weeks can significantly reduce the swelling of the knee joint in the treatment group,make the articular cartilage tissue redder stained,increase the number of chondrocytes and cartilage thickness,and display a smoother cartilage surface.It shows inhibition of osteoclasts in subchondral bone in TRAP staining in the combined administration group.Conclusion:Our results indicate that the combined administration of Atorvastatin and glucosamine is a potential therapeutic agent for protection of articular cartilage against OA progression.