title: MiR-29a and MiR-140 Protect Chondrocytes against the Anti-Proliferation and Cell Matrix Signaling Changes by IL-1β.
abstract: As a degenerative joint disease, osteoarthritis (OA) constitutes a major cause of disability that seriously affects the quality of life of a large population of people worldwide. However, effective treatment that can successfully reverse OA progression is lacking until now. The present study aimed to determine whether two small non-coding RNAs miR-29a and miR-140, which are significantly down-regulated in OA, can be applied together as potential therapeutic targets for OA treatment. MiRNA synergy score was used to screen the miRNA pairs that potentially synergistically regulate OA. An in vitro model of OA was established by treating murine chondrocytes with IL-1β. Transfection of miR-29a and miR-140 via plasmids was investigated on chondrocyte proliferation and expression of nine genes such as ADAMTS4, ADAMTS5, ACAN, COL2A1, COL10A1, MMP1, MMP3, MMP13 and TIMP metal-lopeptidase inhibitor 1 (TIMP1). Western blotting was used to determine the protein expression level of MMP13 and TIMP1, and ELISA was used to detect the content of type II collagen. Combined use of miR-29a and miR-140 successfully reversed the destructive effect of IL-1β on chondrocyte proliferation, and notably affected the MMP13 and TIMP1 gene expression that regulates extracellular matrix. Although co-transfection of miR-29a and miR-140 did not show a synergistic effect on MMP13 protein expression and type II collagen release, but both of them can significantly suppress the protein abundance of MMP13 and restore the type II collagen release in IL-1β treated chondrocytes. Compared with single miRNA transfection, cotransfection of both miRNAs exceedingly abrogated the suppressed the protein production of TIMP1 caused by IL-1β, thereby suggesting potent synergistic action. These results provided novel insights into the important function of miRNAs' collaboration in OA pathological development. The reduced MMP13, and enhanced TIMP1 protein production and type II collagen release also implies that miR-29a and miR-140 combination treatment may be a possible treatment for OA.
authors.affiliation: Department of Pharmacy, the Second Affiliated Hospital of Harbin Medical University, Harbin, China.
authors.forename: Xianghui
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Orthopedics, the First Affiliated Hospital of Harbin Medical University, Harbin, China.
authors.forename: Zhilei
authors.initials: Z
authors.lastname: Zhen
authors.affiliation: Department of Orthopedics, the First Affiliated Hospital of Harbin Medical University, Harbin, China.
authors.forename: Guodong
authors.initials: G
authors.lastname: Tang
authors.affiliation: Department of Orthopedics, the First Affiliated Hospital of Harbin Medical University, Harbin, China.
authors.forename: Chong
authors.initials: C
authors.lastname: Zheng
authors.affiliation: Department of Orthopedics, the First Affiliated Hospital of Harbin Medical University, Harbin, China.
authors.forename: Guofu
authors.initials: G
authors.lastname: Yang
mesh_terms: D051722:ADAM Proteins / Q000235:genetics / Q000378:metabolism; D000071121:ADAMTS4 Protein; D053674:Aggrecans / Q000235:genetics / Q000378:metabolism; D000818:Animals; D000833:Animals, Suckling; D049109:Cell Proliferation / Q000187:drug effects; D019902:Chondrocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D024121:Collagen Type X / Q000235:genetics / Q000378:metabolism; D005109:Extracellular Matrix / Q000187:drug effects* / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D053583:Interleukin-1beta / Q000494:pharmacology*; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D053509:Matrix Metalloproteinase 13 / Q000235:genetics / Q000378:metabolism; D051379:Mice; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008954:Models, Biological; D010003:Osteoarthritis / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D061251:Primary Cell Culture; D011348:Procollagen N-Endopeptidase / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics / Q000378:metabolism
chemical_list: C508320:Acan protein, mouse; D053674:Aggrecans; C570620:Col10a1 protein, mouse; C447413:Col2a1 protein, mouse; D024043:Collagen Type II; D024121:Collagen Type X; D053583:Interleukin-1beta; D007527:Isoenzymes; C547828:MIRN140 microRNA, mouse; C547014:MIRN29 microRNA, mouse; D035683:MicroRNAs; C560825:Timp1 protein, mouse; D019715:Tissue Inhibitor of Metalloproteinase-1; D051722:ADAM Proteins; D053509:Matrix Metalloproteinase 13; C507081:Mmp13 protein, mouse; D011348:Procollagen N-Endopeptidase; D000071121:ADAMTS4 Protein
keywords: miR-140; miR-29a; microRNA; osteoarthritis