Satellite cells (SCs) are multipotent skeletal muscle precursor cells, which can lead to muscle regeneration, a crucial process for the treatment of stress urinary incontinence (SUI). The activation of SCs is likely to be caused by inducible nitric oxide synthase (iNOS). This study examines the underlying mechanism of iNOS in SC activation, and the implications of a potential treatment of SUI.
SCs were isolated from the levator ani muscles of rats, followed by measurement of iNOS, paired box gene 7 (Pax7) and myosin D (MyoD) protein expression. Flow cytometric and cell counting kit-8 assays were conducted to assess SC proliferation, apoptosis, cell cycle distribution. SC myotube formation was observed under a light microscope. A rat model of SUI was established using vaginal dilation and bilateral ovarian castration, followed by assessment of leaking point pressure (LPP) and a sneezing test. The levels of MyoD, Pax7, hepatocyte growth factor (HGF) and p-38 in the levator ani muscles were detected by immunohistochemical and western blot analyses.
SCs successfully isolated from rat levator ani muscles showed increased MyoD and reduced Pax7 expression. Upregulation of iNOS promoted differentiation and myotube formation of SCs. iNOS elevated the expression of HGF, which in turn activated the p38/MAPK signaling pathway, promoting the differentiation of SCs. SCs overexpressing iNOS increased LPP, thus preventing SUI in an in vivo rat model.
Our results show that iNOS could activate the HGF-dependent p38/MAPK signaling pathway to alleviate SUI, potentially providing a novel therapeutic strategy for SUI.
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Stress urinary incontinence
Inducible nitric oxide synthase
Hepatocyte growth factor
Jun amino-terminal kinase
Mitogen-activated protein kinase
Dulbecco's modified Eagle's medium
Fetal bovine serum
Polymerase chain reaction
Small interfering RNA
Cell counting kit-8
Reverse transcription quantitative polymerase chain reaction
Paired box gene 7
Tris-buffered saline with Tween-20
Hematoxylin and eosin
PBS containing 0.1% Tween-20
Maximum bladder capacity
Leaking point pressure
Analysis of variance
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The authors would like to acknowledge the helpful comments on this paper received from the reviewers.
This study was supported by the Yunnan Provincial Science and Technology Department (No. 2015FA007); National Natural Science Foundation of China (81860127); Medical Discipline Leaders of Health and Family Planning Commission of Yunnan Province (D-201615); Young and Middle-Aged Academic and Technical Leaders Reserved Scholar of Yunnan Province (No. 2017HB038); the Engineering and Research Center of Yunnan College and University for Female Pelvic Floor Disease Diagnosis and Treatment.
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Guan, F., Wu, J., Li, J. et al. Inducible nitric oxide synthase promotes differentiation of satellite cells and prevents stress urinary incontinence via HGF-mediated p38/MAPK signaling. World J Urol (2020). https://doi.org/10.1007/s00345-020-03289-7
- Stress urinary incontinence
- Inducible nitric oxide synthase
- Satellite cells
- Hepatocyte growth factor
- p38 mitogen-activated protein kinase signaling pathway