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Effects of mechanical trauma on the differentiation and ArfGAP3 expression of C2C12 myoblast and mouse levator ani muscle

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Abstract

Introduction and hypothesis

Severe mechanical injury or inadequate repair of the levator ani muscle (LAM) is a key contributor to the development of pelvic floor dysfunction (PFD). We explored the effects of mechanical stress on myoblasts and LAM at the cellular and animal level and the possible mechanism of PFD induced by mechanical trauma.

Methods

A C2C12 cell mechanical injury model was established with a four-point bending device, and a LAM injury mouse model was established via vaginal distention and distal traction, a common way of simulating the birth injury. The cells were divided into control, 1333 μ strain for 4-h cyclic mechanical strain (CMS), 1333 μ strain for 8-h CMS, and 5333 μ strain for 4-h CMS groups. Mice were divided into control and injury groups. After treatment, mitochondrial membrane potential (ΔΨm), reactive oxygen species (ROS) levels, indicators of oxidative damage, cell apoptosis, muscle and cell morphology, cell differentiation, and expression of adenosine diphosphate (ADP)-ribosylation factor GTPase activating protein 3 (ArfGAP3) were detected.

Results

5333 μ strain for 4-h CMS loading could induce myoblast injury with a reduction of ΔΨm, increased ROS levels, aggravation of oxidative damage-associated proteins NADPH oxidase 2 (NOX2) and xanthine oxidase (XO), and an increased apoptosis rate of C2C12 cells. At the same time, the injury CMS loading can promote the differentiation of myoblasts and increase the expression of ArfGAP3, a factor regulating intracellular transport. Mechanical trauma could also lead to the oxidative damage of LAM, indicated by 8-hydroxy-2′-deoxyguanosine(8-OHdG), NOX2 and XO protein accumulation, and increase the expression of ArfGAP3 in LAM.

Conclusions

Oxidative stress caused by mechanical trauma induces dysfunction and damage repairing of LAM and C2C12 myoblast, and ArfGAP3 may promote the repairing process.

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Abbreviations

PFD:

Pelvic floor dysfunction

SUI:

Stress urinary incontinence

POP:

Pelvic organ prolapse

LAM:

Levator ani muscle

CMS:

Cyclic mechanical strain

ΔΨm:

Mitochondrial membrane potential

MyHC:

Myosin heavy chain

ArfGAP3:

Adenosine diphosphate-ribosylation factor GTPase activating protein 3

NOX2:

NADPH oxidase 2

XO:

Xanthine oxidase

8-OHdG:

8-hydroxy-2′-deoxyguanosine

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (no. 81771562; no. 81971364). The authors would like to thank all the teachers in the Department of Gynecology and Obstetrics and Central Laboratory, Renmin Hospital of Wuhan University, for their technical assistance.

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Renmin Hospital of Wuhan University, #238 Jiefang Road, Wuhan, 430060, Hubei Province, People’s Republic of China

    Yingyi Yi, Linlin Wang, Suting Li, Bingshu Li, Cheng Liu & Li Hong

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  1. Yingyi Yi
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  2. Linlin Wang
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  3. Suting Li
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  4. Bingshu Li
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  5. Cheng Liu
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  6. Li Hong
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Contributions

YY: study design, protocol development, data curation, formal analysis, and manuscript writing; LW: mouse modeling, project administration, data collection, and analysis; SL: tissue sampling of mice and data collection; BL: molecular experiment, data collection, and analysis; CL: cell experiment, data collection, and analysis; LH: conceptualization, funding acquisition, and final approval.

Corresponding author

Correspondence to Li Hong.

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Ethical approval

The procedures of the animal study received approval from the ethics committee of the Institutional Animal Care and Use Committee of Renmin Hospital of Wuhan University (20140305).

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Yi, Y., Wang, L., Li, S. et al. Effects of mechanical trauma on the differentiation and ArfGAP3 expression of C2C12 myoblast and mouse levator ani muscle. Int Urogynecol J 31, 1913–1924 (2020). https://doi.org/10.1007/s00192-019-04212-4

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  • DOI: https://doi.org/10.1007/s00192-019-04212-4

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