锌指蛋白 440 促进人关节突和膝关节骨关节炎软骨细胞的软骨退行性病变机制。
Zinc finger protein-440 promotes cartilage degenerative mechanisms in human facet and knee osteoarthritis chondrocytes.
机构信息
Schroeder Arthritis Institute, Krembil Research Institute, University Health Network, Toronto, Ontario, Canada; Institute of Medical Science, Department of Medicine, University of Toronto, Ontario, Canada; Department of Rheumatology, University of Toronto, Ontario, Canada.
Schroeder Arthritis Institute, Krembil Research Institute, University Health Network, Toronto, Ontario, Canada; Department of Surgery, University of Toronto, Ontario, Canada.
出版信息
Osteoarthritis Cartilage. 2021 Mar;29(3):372-379. doi: 10.1016/j.joca.2020.12.004. Epub 2020 Dec 24.
OBJECTIVES
To investigate the role of zinc finger protein 440 (ZNF440) in the pathophysiology of cartilage degeneration during facet joint (FJ) and knee osteoarthritis (OA).
METHODS
Expression of ZNF440 in FJ and knee cartilage was determined by immunohistochemistry, quantitative (q)PCR, and Western blotting (WB). Human chondrocytes isolated from FJ and knee OA cartilage were cultured and transduced with ZNF440 or control plasmid, or transfected with ZNF440 or control small interfering RNA (siRNA), with/without interleukin (IL)-1β. Gene and protein levels of catabolic, anabolic and apoptosis markers were determined by qPCR or WB, respectively. In silico analyses were performed to determine compounds with potential to inhibit expression of ZNF440.
RESULTS
ZNF440 expression was increased in both FJ and knee OA cartilage compared to control cartilage. In vitro, overexpression of ZNF440 significantly increased expression of MMP13 and PARP p85, and decreased expression of COL2A1. Knockdown of ZNF440 with siRNA partially reversed the catabolic and cell death phenotype of human knee and FJ OA chondrocytes stimulated with IL-1β. In silico analysis followed by validation assays identified scriptaid as a compound with potential to downregulate the expression of ZNF440. Validation experiments showed that scriptaid reduced the expression of ZNF440 in OA chondrocytes and concomitantly reduced the expression of MMP13 and PARP p85 in human knee OA chondrocytes overexpressing ZNF440.
CONCLUSIONS
The expression of ZNF440 is significantly increased in human FJ and knee OA cartilage and may regulate cartilage degenerative mechanisms. Furthermore, scriptaid reduces the expression of ZNF440 and inhibits its destructive effects in OA chondrocytes.
目的
研究锌指蛋白 440（ZNF440）在小关节（FJ）和膝骨关节炎（OA）关节软骨退变中的病理生理学作用。
方法
通过免疫组织化学、定量（q）PCR 和 Western blot（WB）检测 FJ 和膝关节软骨中 ZNF440 的表达。从小关节和膝骨关节炎软骨中分离培养人软骨细胞，转染 ZNF440 或对照质粒，或用 ZNF440 或对照小干扰 RNA（siRNA）转染，有/无白细胞介素（IL）-1β。分别通过 qPCR 或 WB 测定分解代谢、合成代谢和凋亡标志物的基因和蛋白水平。通过计算机分析确定具有抑制 ZNF440 表达潜力的化合物。
结果
与对照软骨相比，FJ 和膝 OA 软骨中 ZNF440 的表达均增加。体外，ZNF440 过表达显著增加了 MMP13 和 PARP p85 的表达，并降低了 COL2A1 的表达。用 siRNA 敲低 ZNF440 可部分逆转 IL-1β刺激的人膝关节和 FJ OA 软骨细胞的分解代谢和细胞死亡表型。计算机分析后验证实验确定 scriptaid 是一种具有潜在下调 ZNF440 表达能力的化合物。验证实验表明，scriptaid 降低了 OA 软骨细胞中 ZNF440 的表达，并同时降低了过表达 ZNF440 的人膝关节 OA 软骨细胞中 MMP13 和 PARP p85 的表达。
结论
ZNF440 在人类 FJ 和膝关节 OA 软骨中的表达显著增加，可能调节软骨退行性机制。此外，scriptaid 降低了 ZNF440 的表达，并抑制了其在 OA 软骨细胞中的破坏性作用。
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