Tianjin Medical Journal

Tianjin Medical Journal ›› 2021, Vol. 49 ›› Issue (9): 921-925.doi: 10.11958/20211562

• Cell and Molecular Biology • Previous Articles     Next Articles

Study on the mechanism of alendronate in dexamethasone induced autophagy in C2C12 cells

TIAN Ai-xian1, MA Jian-xiong2, MA Xin-long2△, LI Yan1   

  1. 1 Department of Biomaterials and Histomorphology, 2 Biomechanics Research Laboratory, Institute of Orthopaedics, Tianjin Hospital (Tianjin Hospital, Tianjin University), Tianjin Key Laboratory of Orthopaedics Biomechanics and Medical Engineering, Tianjin 300050, China
  • Received:2021-07-02 Revised:2021-07-26 Published:2021-09-15 Online:2021-09-18

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Abstract: Abstract: Objective　To investigate the role and mechanism of alendronate (ALN) in dexamethasone (Dexa) induced autophagy in C2C12 cells. Methods　C2C12 cells were divided into the control group (DMSO treatment), Dexa group (100 μmol/L Dexa treatment) and Dexa+ALN group (100 μmol/L Dexa+1.0 μmol/L ALN treatment). C2C12 cells were treated with 0, 0.1, 0.5 and 1.0 μmol/L ALN for 48 h, respectively. The proliferation level of C2C12 cells was detected by CCK-8 method, and the appropriate dose of ALN was screened. The differentiation of C2C12 cells was detected by hematoxylin-eosin (HE) staining. The diameter and fusion index of myotubule were measured by Image J. Western blot assay was used to detect the expression levels of myoglobin MHC, MuRF1 and autophagy related proteins LC3 and Beclin-1 in C2C12 cells. Immunofluorescence was used to detect the myosin heavy chain (MHC) expression of C2C12 cells. Results　The appropriate dose of ALN was 1.0 μmol/L for subsequent experiments. HE staining and Image J showed that the diameter and fusion index of myotubule were significantly increased when the dose of ALN was 1.0 μmol/L (P＜0.01). Western blot results showed that compared with the control group, MuRF1 protein increased in Dexa group, while the expression of MuRF1 protein was significantly down-regulated in Dexa+ALN group than that of Dexa group, and autophagy related proteins LC3 and Beclin-1 increased (P＜0.01). Immunofluorescence results showed that compared with the control group, the fluorescence expression intensity (red light) of MHC protein was significantly decreased in Dexa group, and the fluorescence expression intensity of MHC protein was significantly increased in Dexa+ALN group than that of Dexa group. Conclusion　The 1.0 μmol/L ALN can effectively promote the proliferation and differentiation of C2C12 cells, improve the inhibitory effect of Dexa on the differentiation of C2C12 cells, and inhibit the expression of myodegradable protein MuRF1. The mechanism may be related to the moderate activation of autophagy signaling pathway.

Key words: diphosphonates, dexamethasone, autophagy, beclin-1, alendronate, C2C12 cell, signaling pathways