Mechanical stretch enhances sex steroidogenesis in C₂C₁₂ skeletal muscle cells

Skeletal muscle contains estrogens and estrogen synthesis-related enzymes. However, it has not been reported whether myoblasts cultured in vitro also express these enzymes. The purpose of the current study was to address these issues and to explore the effects of mechanical stretch on the enzyme system. The in vitro cultured C₂C₁₂ mouse myoblasts were divided into the control, stretch, testosterone and stretch plus testosterone groups. Cells in the stretch and stretch plus testosterone groups were mechanically stretched with the Flexercell cell stress loading device at an amplitude of 10% and in a frequency of 0.5 Hz for 8 h. Cells in the testosterone and stretch plus testosterone groups were incubated with 100 nM testosterone for 24 h before distraction. Following the treatments, cell proliferation and estradiol levels, as well as the expressions of 17β-hydroxysteroid (17β-HSD), 3β-hydroxysteroid (3β-HSD) and aromatase were analyzed. Compared to the control, the cell proliferation in all experimental groups increased significantly, the estradiol levels in the mechanically stretched groups were significantly higher, and, moreover, the estradiol levels were positively correlated with the cell proliferation (r = 0.615, p < 0.01). Additionally, analyses of aromatase protein and mRNA showed that, compared to the control, their levels were significantly increased upon stretching and testosterone exposure. Similarly, the protein and mRNA levels of both 3β-HSD and 17β-HSD in the stretched cells differed significantly from the control. In the presence of aromatase and 5α-reductase inhibitors, the protein and mRNA levels of these enzymes altered significantly compared to the control. Conclusions: Steroid synthases were detected in the C₂C₁₂ myoblasts cultured in vitro, the synthesized estrogen was closely related to the cell proliferation, and mechanical stretch was the external factor that affected the expression of the estrogen synthesis-related enzymes.