The mechanism of microRNA-153-3p/snca axis promoting osteoporosis by regulating osteoclast differentiation
Objective To investigate how the interactions between microR NA(miR)-153-3p andα-synuclein(SNCA)contribute to osteoporosis in mice.Methods The moust osteoporosis(OVX)model was constructed by bilateral ovariectomy,and the pathological changes were confirmed by hematoxylin-eosin(HE)staining.Real-time fluorescent quantitative reverse transcription polymerase chain reaction(RT-qPCR)and Western blotting were used to quantify the messenger RNA(mRNA)and protein expres-sion of target genes.After treating mouse RAW264.7 cells with receptor activator of nuclear factor-KB lig-and(RANKL),morphological changes were observed under the microscopy.Methyl thiazolyl tetrazolium(MTT)assay was used to evaluate cell viability,Transwell experiment was used to detect osteoclast migration ability,and dual-luciferase reporter(DLR)assay was used to verify the interaction between miR-153-3p and snca.Results The expression of snca was low in OVX mice(t=11.436,P＜0.05),while the expres-sion of miR-153-3p was high(t=15.833,P＜0.05).Inhibiting miR-153-3p or overexpressing snca sup-pressed osteoporosis.The expression of snca was low in RANKL-induced RAW264.7 cells(t=11.796,P＜0.05),while the expression of miR-153-3p was high(t=10.448,P＜0.05).Overexpression of snca inhibited osteoclast differentiation,evidenced by the expression changes in a series of osteoclast markers[tartrate resistant acid phosphatase(TRAP):t=7.493,P＜0.05;Cathepsin K:t=9.536,P＜0.05;matrix metalloproteinase(MMP)-9:t=20.371,P＜0.05;MMP-2:t=31.924,P＜0.05].miR-153-3p promoted osteoclast differentiation by inhibiting snca expression(F=123.390,P＜0.05),and overexpres-sion of snca restored the effect of overexpressing miR-153-3p(F=136.515,P＜0.05).DLR assay showed that miR-153-3p negatively regulated the expression of snca(F=92.528,P＜0.05).Conclusion Inhibition of miR-153-3p suppresses the function of osteoclasts by promoting snca expression,and then in-hibits the process of osteoporosis in mice.
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