Therapeutic Effect of AAV8-Mediated miR-23a in Immobilization-Induced Muscle Atrophy in Mice

Lu Na; Wang Yijin; Wenxin .; Farra Aidah Jumuddin

doi:10.31632/ijalsr.2024.v07i03.008

Lu Na Hainan Medical University, Hainan, 570102,China & Lincoln University College, Wisma Lincoln, 12-18, Jalan SS 6/12, 47301 Petaling Jaya, Selangor, Malaysia https://orcid.org/0009-0004-9483-8767
Wang Yijin Emergency and Trauma College of Hainan Medical University, Hainan, 570102, China https://orcid.org/0009-0003-1652-8934
Wenxin . Hainan Medical University, Hainan, 570102,China https://orcid.org/0009-0004-6648-683X
Farra Aidah Jumuddin Faculty of Medicine，Lincoln University College , Wisma Lincoln, No. 12-18, Jalan SS 6/12, 47301 Petaling Jaya, Selangor Darul Ehsan, Malaysia https://orcid.org/0000-0002-3817-7050

DOI https://doi.org/10.31632/ijalsr.2024.v07i03.008

Abstract

Objective: To investigate the therapeutic effect of miR-23a in immobilization-induced muscle atrophy in mice. Methods: (Experiment 1) Twelve C57BL6 wild-type mice were randomly divided into two groups: Sham group (sham surgery) and Immobilization (hind limb immobilization surgery). Fluorescence quantitative PCR was performed to detect the expression changes of miR-23a, MuRF-1, and Atrogin-1 7 days post-surgery. (Experiment 2) Twelve C57BL6 wild-type mice were randomly divided into 4 groups according to surgery type (sham surgery and hind limb immobilization surgery) and injection type (AAV8 or AAV8-OEmiR-23a), fluorescence quantitative PCR to detect changes in MuRF-1 and Atrogin-1 expression in mouse gastrocnemius muscle. (Experiment 3) NFATc3 expression changes were detected by fluorescence quantitative PCR 48 hours post-transfection. (Experiment 4) Twelve C57BL6 wild-type mice underwent hind limb immobilization surgery, divided into 4 groups : Immobilization+AAV8-control, Immobilization+AAV8-OEmiR23a, Immobilization+AAV8-shNFATc3, and Immobilization+AAV8-OEmiR-23a+AAV8-shNFATc3. quantitative PCR to detect changes in MuRF-1 and Atrogin-1 expression in mouse gastrocnemius muscle. Results: Fluorescence quantitative PCR results showed that miR-23a was downregulated in immobilization-induced muscle atrophy, while MuRF-1 and Atrogin-1 expression was upregulated. WGA staining results showed that intramuscular injection of AAV8-OEmiR-23a could significantly alleviate the decrease in muscle fiber cross-sectional area and the increase in expression of muscle atrophy marker genes MuRF-1 and Atrogin-1 caused by hind limb immobilization, Fluorescence quantitative PCR revealed that when miR-23a expression was inhibited, NFATc3 expression was downregulated; when miR-23a was overexpressed, NFATc3 expression was upregulated, Fluorescence quantitative PCR results showed that intramuscular injection of AAV8-OEmiR-23a and AAV8-shNFATc3 could not alleviate the increase in expression of muscle atrophy marker genes MuRF-1 and Atrogin-1 caused by hind limb immobilization. Conclusion: miR-23a can treat hind limb immobilization-induced muscle atrophy in mice.

Keywords: miR-23a, hind limb immobilization-induced muscle atrophy, NFATc3

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Therapeutic Effect of AAV8-Mediated miR-23a in Immobilization-Induced Muscle Atrophy in Mice

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