Wnt 5a/Smad 2/3 Activation and Sclerostin Inhibition in Subchondral Bone Cells Promote Trabecular Sclerosis in Osteoarthritis

doi:10.3870/j.issn.1672-8009.2025.06.003

Abstract

Abstract: Objective To investigate the effect of Wnt 5a/Smad 2/3 activation and sclerostin inhibition in subchondral bone cells on trabecular sclerosis in osteoarthritis. Methods The co-cultured system of chondrocytes and osteoblasts was established.Cells were divided into 5 groups:Control group,OE-vector group,LIF OE group,shRNA-NC group and shRNA-LIF group.RNA immunoprecipitation(RIP)was used to detect the direct interaction between LIFR and sclerostin encoding gene SOST mRNA.The relative expression levels of SOST mRNA was detected by qRT-PCR.LIF expression in chondrocytes was overexpressed or knocked-down,and the relative expression levels of LIF and LIFR was detected by Western blotting.fourty male C57BL 6J mice aged 7-8 weeks were randomly divided into 7 groups:Control group,Model group,Sham group with 4-week or 8-week induction,and Model+LIFR antagonist group with 8-week induction.Micro-CT was used to measure the bone volume fraction(BV/TV),trabecular thickness(Tb.Th)and trabecular number(Tb.N).HE staining and safranin O-fast green staining were used to analyze the histopathological changes in mice joint tissues.The relative expression levels of Wnt 5a,Smad 2/3,Sclerostin,leukemia inhibitory factor(LIF)/LIF receptor(LIFR),tartrate-resistant acid phosphatase(TRAP)and matrix metalloproteinases 13(MMP-13)were detected by Western blotting. Results RIP results showed that LIFR and SOST mRNA had direct interaction.Overexpression of LIF increased LIFR expression levels(P<0.05)and decreased Sclerostin expression levels(P<0.05);knocking down LIF produced the opposite results.After 8 weeks of induction,the BV/TV and Tb.Th were increased(P<0.05),and the Tb.N was decreased(P>0.05)in the Model group when compared with those in the Sham group.Compared with those in the Model group,the BV/TV and Tb.Th were decreased(P<0.05)in the Model+LIFR antagonist group,and the Tb.N was increased(P>0.05).HE staining and Safranin O-fast green cartilage staining results showed that in the Model group,cartilages was destroyed,the intercellular matrix degraded,cell arrangement was disrupted,the cartilage layer disintegrated,and the subchondral bone was affected.In the Model+LIFR antagonist group,the cartilage surfaces area and proliferation area were destroyed,but the hypertrophy zone remained relatively intact,suggesting that cartilage zone destruction was relatively alleviated following LIFR antagonist treatment.After 4 weeks or 8 weeks of induction,the expression levels of Wnt 5a,p-Smad 2/3 in the Model group were increased(P<0.05),the expression levels of sclerostin was decreased(P<0.05),and the expression levels of LIF,LIFR,TRAP and MMP-13 were up-regulated(P<0.05)when compared with those in the Sham group. Conclusion Wnt 5a/Smad 2/3 activation and sclerostin inhibition in subchondral bone cells promote trabecular sclerosis.

Key words: osteoarthritis, subchondral bone, trabecular sclerosis, sclerostin, leukemia inhibitory factor

CLC Number:

R684.3
R332

Ekermujiang Arken, HAN Xiaoping, CUI Yong, HUANG Tao. Wnt 5a/Smad 2/3 Activation and Sclerostin Inhibition in Subchondral Bone Cells Promote Trabecular Sclerosis in Osteoarthritis[J]. Journal of Medical Molecular Biology, 2025, 22(6): 548-556.

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