Overexpression of miR-148a-3p inhibits extracellular matrix degradation and alleviates IL-1β-induced intervertebral disc degeneration

Document Type : Original Article


1 Department of Orthopaedic Surgery, The Fifth Affiliated Hospital of Wenzhou Medical University, Affiliated Lishui Hospital of Zhejiang University, Lishui Municipal Central Hospital, 289 Kuocang Road, Lishui 323000, Zhejiang Province, China

2 Lishui Second People’s Hospital, Beihuan Road 69, Liandu District, Lishui323000, Zhejiang Province, China


Objective(s): Recently, studies on microRNAs (miRNAs) and their targets and related genes have provided new therapeutic opportunities for controlling intervertebral disc degeneration (IDD). We aimed to investigate the effects of miR-148a-3p overexpression on IDD progression.
Materials and Methods: This study used microRNA microarrays to analyze key regulators of IDD. Q-PCR was used to verify the IL-1β-induced down-regulation of miR-148a-3p expression both in nucleus pulposus (NP) tissues of IDD patients and in degenerated NP cells (NPCs) of rats. Rat NPC micromass cultures and ex vivo intervertebral disc (IVD) culture models were established, and histological staining was performed to verify the effect of miR-148a-3p on the general morphology and proteoglycan and collagen contents of IVDs. In addition, q-PCR and western blotting analyses were performed to examine the expression of ECM molecules and matrix-degrading enzymes. A luciferase reporter assay was used to confirm the target genes of miR-148a-3p.
Results: Our data revealed that miR-148a-3p was down-regulated in IDD. Overexpression of miR-148a-3p had no effect on ACAN or COL2A1 gene expression but decreased MMP3, MMP13, and ADAMTS5 gene expression. The matrix deposited by miR-148a-3p-overexpressing rat NPCs contained high levels of proteoglycans and collagen. The ex vivo experiments verified that agomiR-148a-3p alleviated the NPC matrix degradation induced by IL-1β. A luciferase reporter assay confirmed that miR-148a-3p directly targeted ADAMTS5 and MMP13.
Conclusion: We proved that miR-148a-3p can attenuate ECM loss and protect NP function by inhibiting matrix-degrading enzymes.


1. Chou R. Low Back Pain. Ann Intern Med 2021; 174:Itc113-itc128.
2. Hartvigsen J, Hancock MJ, Kongsted A, Louw Q, Ferreira ML, Genevay S, et al. What low back pain is and why we need to pay attention. Lancet 2018; 391: 2356-2367.
3. Livshits G, Popham M, Malkin I, Sambrook PN, Macgregor AJ, Spector T, et al. Lumbar disc degeneration and genetic factors are the main risk factors for low back pain in women: the UK Twin Spine Study. Ann Rheum Dis 2011; 70: 1740-1745.
4. Guerrero J, Häckel S, Croft AS, Hoppe S, Albers CE, Gantenbein B. The nucleus pulposus microenvironment in the intervertebral disc: the fountain of youth? Eur Cell Mater 2021; 41: 707-738.
5. Choi H, Johnson ZI, Risbud MV. Understanding nucleus pulposus cell phenotype: a prerequisite for stem cell based therapies to treat intervertebral disc degeneration. Curr Stem Cell Res Ther 2015; 10: 307-316.
6. Tu J, Li W. Single-cell transcriptome profiling reveals multicellular ecosystem of nucleus pulposus during degeneration progression. Adv Sci (Weinh) 2021;9: e2103631.
7. Fontana G, See E, Pandit A. Current trends in biologics delivery to restore intervertebral disc anabolism. Adv Drug Deliv Rev 2015; 84: 146-158.
8. Clouet J, Fusellier M, Camus A, Le Visage C, Guicheux J. Intervertebral disc regeneration: From cell therapy to the development of novel bioinspired endogenous repair strategies. Adv Drug Deliv Rev 2019; 146: 306-324.
9. Mori MA, Ludwig RG, Garcia-Martin R, Brandão BB, Kahn CR. Extracellular miRNAs: From biomarkers to mediators of physiology and disease. Cell Metab 2019; 30: 656-673.
10. Wang XQ, Tu WZ, Guo JB, Song G, Zhang J, Chen CC, et al. A bioinformatic analysis of microRNAs’ role in human intervertebral disc degeneration. Pain Med 2019; 20: 2459-2471.
11.Liu P, Chang F, Zhang T, Gao G, Yu C, Ding SQ, et al. Down-regulation of microRNA-125a is involved in intervertebral disc degeneration by targeting pro-apoptotic Bcl-2 antagonist killer 1. Iran J Basic Med Sci 2017; 20: 1260-1267.
12.Gu SX, Li X, Hamilton JL, Chee A, Kc R, Chen D, et al. MicroRNA-146a reduces IL-1 dependent inflammatory responses in the intervertebral disc. Gene 2015; 555: 80-87.
13. Jie J, Xu X, Li W, Wang G. Regulation of apoptosis and inflammatory response in interleukin-1β-induced nucleus pulposus cells by miR-125b-5p via targeting TRIAP1. Biochem Genet 2021; 59: 475-490.
14. Chen X, Li Z, Xu D, Li S. LINC01121 induced intervertebral disc degeneration via modulating miR-150-5p/MMP16 axis.  2020; 22: e3231.
15.Rupaimoole R, Slack FJ. MicroRNA therapeutics: towards a new era for the management of cancer and other diseases. Nat Rev Drug Discov 2017; 16: 203-222.
16. Zhao B, Yu Q, Li H, Guo X, He X. Characterization of microRNA expression profiles in patients with intervertebral disc degeneration. Int J Mol Med 2014; 33: 43-50.
17. Li G, Tang X, Chen H, Sun W, Yuan F. miR-148a inhibits pro-inflammatory cytokines released by intervertebral disc cells by regulating the p38/MAPK pathway. Exp Ther Med 2018; 16: 2665-2669.
18. Oh CD, Im HJ, Suh J, Chee A, An H, Chen D. Rho-Associated Kinase Inhibitor Immortalizes Rat Nucleus Pulposus and Annulus Fibrosus Cells: Establishment of Intervertebral Disc Cell Lines With Novel Approaches. Spine (Phila Pa 1976) 2016; 41: E255-261.
19. Rustenburg CM, Snuggs JW, Emanuel KS, Thorpe A, Sammon C, Le Maitre CL, et al. Modelling the catabolic environment of the moderately degenerated disc with a caprine ex vivo loaded disc culture system. Eur Cell Mater 2020; 40: 21-37.
20. Chiba K, Andersson GB, Masuda K, Momohara S, Williams JM, Thonar EJ. A new culture system to study the metabolism of the intervertebral disc in vitro. Spine (Phila Pa 1976) 1998; 23: 1821-1828. 
21. Risbud MV, Shapiro IM. Role of cytokines in intervertebral disc degeneration: pain and disc content. Nat Rev Rheumatol 2014; 10: 44-56.
22. Phillips KL, Jordan-Mahy N, Nicklin MJ, Le Maitre CL. Interleukin-1 receptor antagonist deficient mice provide insights into pathogenesis of human intervertebral disc degeneration. Ann Rheum Dis 2013; 72: 1860-1867.
23. Liu ZQ, Fu WQ, Zhao S, Zhao X. Regulation of insulin-like growth factor 1 receptor signaling by microRNA-4458 in the development of lumbar disc degeneration. Am J Transl Res 2016; 8: 2309-2316.
24. Yan N, Yu S, Zhang H, Hou T. Lumbar Disc Degeneration is facilitated by miR-100-mediated FGFR3 suppression. Cell Physiol Biochem 2015; 36: 2229-2236.
25. Song YQ, Karasugi T, Cheung KM, Chiba K, Ho DW, Miyake A, et al. Lumbar disc degeneration is linked to a carbohydrate sulfotransferase 3 variant. J Clin Invest 2013; 123: 4909-4917.
26. Adams BD, Parsons C, Walker L, Zhang WC, Slack FJ. Targeting noncoding RNAs in disease. J Clin Invest 2017; 127: 761-771.
27. Bedene A, Mencej Bedrač S, Ješe L, Marc J, Vrtačnik P, Preželj J, et al. MiR-148a the epigenetic regulator of bone homeostasis is increased in plasma of osteoporotic postmenopausal women.  2016; 128: 519-526.
28. Liu J, Dang L, Li D, Liang C, He X, Wu H, et al. A delivery system specifically approaching bone resorption surfaces to facilitate therapeutic modulation of microRNAs in osteoclasts. Biomaterials 2015; 52: 148-160.
29. Liu H, Su H, Wang X, Hao W. MiR-148a regulates bone marrow mesenchymal stem cells-mediated fracture healing by targeting insulin-like growth factor 1. J Cell Biochem 2018; 120: 1350-1361.
30. Jones TL, Esa MS, Li KHC, Krishnan SRG, Elgallab GM, Pearce MS, et al. Osteoporosis, fracture, osteoarthritis & sarcopenia: A systematic review of circulating microRNA association. Bone 2021; 152: 116068.
31. Vonk LA, Kragten AH, Dhert WJ, Saris DB, Creemers LB. Overexpression of hsa-miR-148a promotes cartilage production and inhibits cartilage degradation by osteoarthritic chondrocytes. Osteoarthritis Cartilage 2014; 22: 145-153.