Abstract:
【Objective】 To investigate the effects of miR-486 on proliferation of sheep(
Ovis aries) skeletal muscle satellite cell and expression level of genes related to PI3K-Akt signal pathway, and finally lay the foundation for illuminating the regulatory mechanism of miR-486 on development of sheep skeletal muscle.【Method】 Here the skeletal muscle satellite cells of one day-old Bashby sheep were applied, and the proliferation of satellite cells and the expression levels of 8 key genes,
PTEN,
GSK3b,
PRKCα,
Raf-1,
MAPK1,
PKN1,
Casp9 and
SGK1 in PI3K-Akt signal pathway were detected when the level of miR-486 in cells were up-or down-regulated by transferring into miR-486 mimics and miR-486 inhibitor.【Result】 The result showed that the skeletal muscle satellite cells in blank control and treated by miR-486 mimics, miR-486 mimics negative control, and miR-486 inhibitor negative control all performed the typical S-shaped proliferation curve, but thespeed of proliferation existed great differenceamong treatments. The proliferation of satellite cells was accelerated when the level of miR-486 was up-regulated, conversely when the level of miR-486 was down-regulated and the cells kept rest state. The results of real-time fluorescence quantitative PCR showed that the expression levels of
PTEN,
Raf-1, and
MAPK1 were extremely significantly down-regulated(
P<0.01, the same below), levels of
PRKCα and
PKN1 were extremely significantly up-regulated, and level of
SGK1gene was also significantly up-regulated(
P<0.05), when the level of miR-486 was up-regulated in satellite cells, but the relative expression levels of
GSK3β and
Casp9 gene did not show significantly different(
P>0.05) no matter the level of miR-486 was up-regulated or down-regulated in satellite cells, maybe these two genes played important roles in essential activity of satellite cells.【Conclusion】 miR-486 can affect the proliferation of sheep skeletal muscle satellite cell by regulating the expression of genes in PI3KAkt signal pathway, and provided a theoretical basis for further investigate the regulation mechanism of miR-486 on skeletal muscle development of sheep and breeding of high-quality mutton sheep breeds.