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Dual role of endoplasmic reticulum stress-ATF-6 activation in autophagy and apoptosis induced by cyclic stretch in myoblast

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A Correction to this article was published on 13 July 2023

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Abstract

Objective: Mandibular growth that is induced by functional appliances is closely associated with skeletal and neuromuscular adaptation. Accumulating evidence has proved that apoptosis and autophagy have a vital role in adaptation process. However, little is known about the underlying mechanisms. This study sought to determine whether ATF-6 is involved in stretch-induced apoptosis and autophagy in myoblast. The study also sought to uncover the potential molecular mechanism. Materials and methods: Apoptosis was assessed by TUNEL and Annexin V and PI staining. Autophagy was detected by transmission electron microscopy (TEM) analysis and immunofluorescent staining for autophagy-related protein light chain 3 (LC3). Real time-PCR and western blot were performed to evaluate the expression level of mRNA and proteins that were associated with endoplasmic reticulum stress (ERS), autophagy and apoptosis. Results: Cyclic stretch significantly decreased the cell viability and induced apoptosis and autophagy of myoblasts time-dependently. Stretching stimuli activated ATF-6 pathway and induced ERS-mediated apoptosis. Moreover, using 4-PBA significantly inhibited ERS-related apoptosis, as well as partially decreasing autophagy. In addition, inhibition of autophagy by 3-MA enhanced apoptosis by affecting the expression of CHOP and Bcl-2. However, it had no obvious effects on ERS-related proteins of GRP78 and ATF-6. More importantly, knockdown ATF-6 effectively weakened apoptosis and autophagy. It did so by regulating the expression of Bcl-2, Beclin1 and CHOP, but not cleaved Caspase-12, LC3II and p62 in stretched myoblast. Conclusion: ATF-6 pathway was activated in myoblast by mechanical stretch. ATF-6 may regulate the process of stretch-induced myoblast apoptosis and autophagy via CHOP, Bcl-2 and Beclin1 signaling.

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Data Availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

Change history

Abbreviations

ATF-6:

activating transcription factor 6.

Bcl-2:

B-cell lymphoma-2.

CHOP:

CCAAT enhance-binding protein homologous protein.

DAPI:

4′,6-Diamidino-2-phenylindole.

ERS:

endoplasmic reticulum stress.

FITC:

Fluorescein isothiocyanate.

GRP78:

Glucose-regulated protein 78KD.

LC3:

autophagy-related protein light chain 3.

MTT:

3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, Thiazolyl Blue Tetrazolium Bromide.

TEM:

transmission electron microscopy.

TUNEL:

Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling.

UPR:

unfolded protein response.

3-MA:

3-Methyladenine.

4-PBA:

4-phenylbutyrate.

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Acknowledgements

The authors thank the laboratory members of Central Laboratory of Affiliated Hospital of Qingdao University for their help and instructions.

Funding

This work was supported by the National Natural Science Foundation of China [grant number 32171303].

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Author notes

  1. Qiang Zhang and Guirong Liu contributed equally to this work and should be considered as co-first authors.

Authors and Affiliations

  1. Department of Orthodontics, The Affiliated Hospital of Qingdao University, Qingdao, China

    Qiang Zhang, Ran Liu, Jun Liu, Xuemin Zeng, Dapeng Ren, Xiao Yan & Xiao Yuan

  2. School of Stomatology, Qingdao University, Qingdao, China

    Qiang Zhang, Jun Liu, Xuemin Zeng, Dapeng Ren, Xiao Yan & Xiao Yuan

  3. Department of Stomatology, Qingdao Municipal Hospital, Qingdao, China

    Guirong Liu

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Contributions

Conceptualization: Yuan X，Yan X; Experiment performance: ZQ，LG and LR; Data collection and analysis: LJ; ZQ，ZX and RD wrote the main manuscript text; ZQ prepared Figs. 1, 2 and 3 and LG prepared Figs. 4, 5 and 6; All authors reviewed the manuscript.

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Correspondence to Xiao Yan or Xiao Yuan.

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Zhang, Q., Liu, G., Liu, R. et al. Dual role of endoplasmic reticulum stress-ATF-6 activation in autophagy and apoptosis induced by cyclic stretch in myoblast. Apoptosis 28, 796–809 (2023). https://doi.org/10.1007/s10495-023-01825-5

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