Research Paper Volume 16, Issue 3 pp 2702—2714

MicroRNA-146a gene transfer ameliorates senescence and senescence-associated secretory phenotypes in tendinopathic tenocytes

Che-Chia Hsu1, *, , Shih-Yao Chen2, *, , Po-Yen Ko1, , Fa-Chuan Kwan1, , Wei-Ren Su1, , I-Ming Jou3,4,5, , Po-Ting Wu1,6,7,8,9, ,

  • 1 Department of Orthopaedics, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan
  • 2 Department of Nursing, College of Nursing, Chung Hwa University of Medical Technology, Tainan, Taiwan
  • 3 Department of Orthopaedics, E-Da Hospital, Kaohsiung, Taiwan
  • 4 School of Medicine, College of Medicine, I-Shou University, Kaohsiung, Taiwan
  • 5 GEG Orthopedic Clinic, Tainan, Taiwan
  • 6 Medical Device Innovation Center, National Cheng Kung University, Tainan, Taiwan
  • 7 Department of Orthopaedics, College of Medicine, National Cheng Kung University, Tainan, Taiwan
  • 8 Department of Biomedical Engineering, National Cheng Kung University, Tainan, Taiwan
  • 9 Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University, Tainan, Taiwan
* Equal contribution

Received: May 30, 2023       Accepted: December 13, 2023       Published: February 2, 2024
How to Cite

Copyright: © 2024 Hsu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Objective: Tendinopathy is influenced by multiple factors, including chronic inflammation and aging. Senescent cells exhibit characteristics such as the secretion of matrix-degrading enzymes and pro-inflammatory cytokines, collectively known as senescence-associated secretory phenotypes (SASPs). Many of these SASP cytokines and enzymes are implicated in the pathogenesis of tendinopathy. MicroRNA-146a (miR-146a) blocks senescence by targeting interleukin-1β (IL-1β) receptor–associated kinase 4 (IRAK-4) and TNF receptor–associated factor 6 (TRAF6), thus inhibiting NF-κB activity. The aims of this study were to (1) investigate miR-146a expression in tendinopathic tendons and (2) evaluate the role of miR-146a in countering senescence and SASPs in tendinopathic tenocytes.

Methods: MiR-146a expression was assessed in human long head biceps (LHB) and rat tendinopathic tendons by in situ hybridization. MiR-146a over-expression in rat primary tendinopathic tenocytes was achieved by lentiviral vector-mediated precursor miR-146a transfer (LVmiR-146a). Expression of various senescence-related markers was analyzed by quantitative reverse transcription polymerase chain reaction (qRT-PCR), immunoblotting and immunofluorescence. MiR-146a expression showed a negative correlation with the severity of tendinopathy in human and rat tendinopathic tendons (p<0.001).

Results: Tendinopathic tenocyte transfectants overexpressing miR-146a exhibited downregulation of various senescence and SASP markers, as well as the target molecules IRAK-4 and TRAF6, and the inflammatory mediator phospho-NF-κB. Additionally, these cells showed enhanced nuclear staining of high mobility group box 1 (HMGB1) compared to LVmiR-scramble-transduced controls in response to IL-1β stimulation.

Conclusions: We demonstrate that miR-146a expression is negatively correlated with the progression of tendinopathy. Moreover, its overexpression protects tendinopathic tenocytes from SASPs and senescence through the IRAK-4/TRAF6/NF-kB pathway.


SASPs: senescence-associated secretory phenotypes; miR-146a: microRNA-146a; IL-1β: interleukin-1β; IRAK-4: IL-1β receptor–associated kinase 4; TRAF6: TNF receptor–associated factor 6; LHB: long head biceps; Ham: hamstring; LVmiR-146a: lentiviral vector-mediated precursor miR-146a transfer; LVmiR-scramble: lentiviral vector-mediated precursor miR-scramble transfer; qRT-PCR: quantitative reverse transcription polymerase chain reaction; p-NF-ĸB: phospho-NF-ĸB:; HMGB1: high mobility group box 1; OA: osteoarthritis; IL-6: interleukin-6; MMP: matrix metalloproteinase; UTR: untranslated region; mRNA: messenger RNA; COX-2: cyclooxygenase-2; SA-β-gal: Senescence β-Galactosidase.