World J Mens Health.  2023 Apr;41(2):363-372. 10.5534/wjmh.210226.

Myocardin Reverses Hypoxia-Inducible Factor-1α Mediated Phenotypic Modulation of Corpus Cavernosum Smooth Muscle Cells in Hypoxia Induced by Cobalt Chloride

• Zhou X
• Luo C
• Fan J
• Gao G
• Wang T
• Zhang H
• Wei A

• KMID: 2540968
• DOI: http://doi.org/10.5534/wjmh.210226

Abstract

Purpose
We aimed to investigate the mechanism of phenotypic transformation of corporal cavernosum smooth muscle cells (CCSMCs) under hypoxic conditions in vitro.
Materials and Methods
In this study, a hypoxia model was established using cobalt chloride (CoCl₂). CCSMCs were treated with different concentrations of CoCl₂ for varying time periods, and cell viability was assessed. Hypoxia-inducible factor-1α (HIF-1α), myocardin (Myocd) and phenotypic markers were detected in the CCSMCs. We also transfected the CCSMCs with si-HIF-1α and Ad-Myocd and evaluated the effects on phenotypic modulation of CCSMCs and the relationship between HIF-1α and Myocd was evaluated.
Results
CoCl₂ inhibited the viability of CCSMCs in a dose- and time-dependent manner, and treatment with 300 µM CoCl₂ for 48 hours were the optimal conditions for establishing the hypoxia model. The results showed increased expression levels of HIF-1α and osteopontin and decreased Myocd, alpha-smooth muscle actin, and calponin levels in CCSMCs under hypoxia. HIF-1α knockdown reversed hypoxia-induced phenotypic transformation with elevated Myocd expression. Overexpression of Myocd also reversed the effect of hypoxia on the phenotypic switch, but did not affect HIF-1α expression.
Conclusions
Our findings showed that HIF-1α was involved in the effect of hypoxia induced by CoCl₂ on CCSMC phenotypic modulation, and Myocd overexpression could inhibit this process. Thus, Myocd might be a potential therapeutic target for erectile dysfunction under hypoxia or HIF-1α activation.

Keyword

Hypoxia; Myocardin; Myocytes; smooth muscle; Phenotype