医学分子生物学杂志2025，Vol.22Issue(6)：548-556,9.DOI:10.3870/j.issn.1672-8009.2025.06.003
骨关节炎软骨下骨细胞中Wnt 5a/Smad 2/3激活及Sclerostin的抑制促进骨小梁硬化
Wnt 5a/Smad 2/3 Activation and Sclerostin Inhibition in Subchondral Bone Cells Promote Trabecular Sclerosis in Osteoarthritis
摘要
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-cul-tured system of chondrocytes and osteoblasts was established.Cells were divided into 5 groups:Con-trol group,OE-vector group,LIFOE group,shRNA-NC group and shRNA-LIF group.RNA immu-noprecipitation(RIP)was used to detect the direct interaction between LIFR and sclerostin enco-ding 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 expres-sion 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 his-topathological 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 phos-phatase(TRAP)and matrix metalloproteinases 13(MMP-13)were detected by Western blot-ting.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 affect-ed.In the Model+LIFR antagonist group,the cartilage surfaces area and proliferation area were de-stroyed,but the hypertrophy zone remained relatively intact,suggesting that cartilage zone destruc-tion was relatively alleviated following LIFR antagonist treatment.After 4 weeks or 8 weeks of induc-tion,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,LI-FR,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.关键词
骨关节炎/软骨下骨/骨小梁硬化/硬化蛋白/白血病抑制因子Key words
osteoarthritis/subchondral bone/trabecular sclerosis/sclerostin/leukemia in-hibitory factor分类
医药卫生引用本文复制引用
艾克热木江·阿尔肯,韩小平,崔泳,黄涛..骨关节炎软骨下骨细胞中Wnt 5a/Smad 2/3激活及Sclerostin的抑制促进骨小梁硬化[J].医学分子生物学杂志,2025,22(6):548-556,9.基金项目
新疆维吾尔自治区自然科学基金(No.2021D01C427) This work was supported by a grant from the Natural Seience Foundation of Xinjiang Uygur Autonomous Region(No.2021D01C427) （No.2021D01C427）
