Department of Spine Surgery, Peking University Shenzhen Hospital, Shenzhen Peking University-The Hong Kong University of Science and Technology Medical Center, Shenzhen, China.
Molecular Laboratory of TCM, Department of Basic Medicine, Liaoning University of TCM, 79 Chongshan East Road, Shenyang, 110032, China.
BMC Complement Altern Med. 2019 Jul 30;19(1):191. doi: 10.1186/s12906-019-2607-4.
Wnt/β-catenin signaling pathway is closely related to osteoarthritis. In our preliminary study, β-catenin conditional activation (cAct) mice that specifically over-express β-catenin gene in cartilage chondrocyte exhibits osteoarthritis-like phenotype in the lumbar disc and knee joint. Therefore, we used the mice to model FJ-OA and test the potential curative effect of Velvet Antler Polypeptide (VAP) on this mice model.
We tested the effect of VAP on β-catenin conditional activation mice, and used Cre negative littermates as controls. Micro-CT, histology and histomorphometry analysis were performed to evaluate the curative effect of VAP on mice facet joint-like phenotype. Expression of β-catenin and collagen II was detected by immunohistochemistry (IHC) and western-blot., MMP13, ADAMTS4 and ADAMTS5 was detected by immunofluorescence (IF). RT-PCR analysis was preformed to detect mRNA expression of cartilage degrading enzymes, such as MMP13, ADAMTS4 and ADAMTS5.
Results of micro-CT (μCT) analysis showed that VAP could partially reverse lumbar disc osteophyte formation observed in β-catenin(ex3) mice. Histology data revealed VAP partially improved facet joint cartilage tissue invades. Histomorphometry analysis showed an increase in total cartilage area after VAP treatment. IHC show that VAP reduced β-catenin protein levels and moderately up-regulated collagen II protein levels. RT-PCR and IF data showed that VAP down-regulated the expression of extracellular matrix synthesis (ECM) degradation enzymes MMP13, ADAMTS4 and ADAMTS5.
Taken together, VAP may modulate ECM by inhibits MMP13, ADAMTS4 and ADAMTS5 via Wnt /β-catenin signaling pathway. Velvet Antler Polypeptide may be a potential medicine for FJ-OA.
Wnt/β-catenin 信号通路与骨关节炎密切相关。在我们的初步研究中，β-catenin 条件性激活（cAct）小鼠在软骨细胞中特异性过表达β-catenin 基因，在腰椎间盘和膝关节中表现出骨关节炎样表型。因此，我们使用这些小鼠模型来模拟 FJ-OA，并测试 Velvet Antler Polypeptide（VAP）对这种小鼠模型的潜在治疗效果。
我们测试了 VAP 对β-catenin 条件性激活小鼠的作用，并使用 Cre 阴性同窝仔鼠作为对照。通过 micro-CT、组织学和组织形态计量学分析来评估 VAP 对小鼠小关节样表型的治疗效果。通过免疫组化（IHC）和 Western blot 检测β-catenin 和胶原 II 的表达，通过免疫荧光（IF）检测 MMP13、ADAMTS4 和 ADAMTS5 的表达。通过 RT-PCR 分析检测软骨降解酶如 MMP13、ADAMTS4 和 ADAMTS5 的 mRNA 表达。
micro-CT（μCT）分析结果表明，VAP 可部分逆转β-catenin(ex3)小鼠中观察到的腰椎间盘骨赘形成。组织学数据显示 VAP 部分改善了小关节软骨组织侵犯。组织形态计量学分析显示 VAP 处理后总软骨面积增加。IHC 显示 VAP 降低了β-catenin 蛋白水平并适度上调了胶原 II 蛋白水平。RT-PCR 和 IF 数据显示，VAP 下调了细胞外基质合成（ECM）降解酶 MMP13、ADAMTS4 和 ADAMTS5 的表达。
综上所述，VAP 可能通过 Wnt/β-catenin 信号通路抑制 MMP13、ADAMTS4 和 ADAMTS5 来调节 ECM。Velvet Antler Polypeptide 可能是 FJ-OA 的一种潜在药物。
