ABSTRACT
Objective:
To investigate the effect of miR-204 on the proliferation and differentiation of
osteoblasts in
osteoporosis mice by
Wnt signaling pathway and its mechanism.
Methods:
Female Kunming
mice were divided into
control group, sham operation group and
osteoporosis group.
Ovariectomy mouse models were established and identified by
bilateral ovariectomy;
Mouse primary
osteoblasts were extracted and identified;
Cells was transfected and detected the miR-204 expression levels; MTT was used to detect the viability of each group of
cells;
Alkaline phosphatase (ALP) activity was detected in each
cell group;
Cell flowmetry was used to detect
apoptosis in each group;
Cell flowmetry was used to detect the activity of
Caspase-3 in each group of
cells; Interaction between miR-204, β-
catenin and LRP-5 was detected by dual
luciferase reporter gene.
Western blot was used to detect the expression of
Wnt signaling pathway-related
proteins.
Results:
The
bone mineral density of the
osteoporosis group was significantly lower than that of the
control group and the sham operation group ( P=0.007, P=0.057) , indicating that the
osteoporosis mice were successfully modeled; The expression level of miR-204 was significantly increased in the miR-204 mimics group ( P=0.007) , and decreased in the miR-204 inhibitor group ( P=0.031) ; The activity of
bone cell and ALP activity of miR-204 mimics increased ( P=0.007, P=0.043) , and the activity of
bone cell and ALP decreased by miR-204 inhibitor ( P=0.007, P=0.035) ; The invasive
ability of miR-204 mimics was significantly increased ( P=0.006) , and the invasive
ability of miR-204 inhibitor was decreased ( P=0.036) ; The
apoptosis ability and
Caspase-3 activity of miR-204 mimics were decreased ( P=0.041, P=0.045) , and the
apoptosis ability and
Caspase-3 activity of
bone cells were enhanced by miR-204 inhibitor ( P=0.005, P=0.039) ; There were targeting relationship between miR-204 and β-
catenin, LRP-5. The expressions of β-
catenin and LRP-5
protein in
osteoblasts of miR-204 mimics were up-regulated ( P=0.043, P=0.009) , and the expression of β-
catenin and LRP-5
protein in
bone cells of miR-204 inhibitor was down-regulated ( P=0.041, P=0.032) .
Conclusion:
miR-204 maybe promote the proliferation and differentiation of
osteoblasts, activate
Wnt signaling pathway, and has certain protective effect on
osteoporosis.