Zinc finger protein A20 regulates the development and progression of osteoarthritis by affecting the activity of NF-κB p65

Abstract

Objective

To investigate the role of Zinc finger protein A20 in osteoarthritis (OA) by regulating NF-κB p65.

Methods

A20, MMP1, MMP13 and IL-1β expressions in human OA cartilage samples were detected by qRT-PCR. IL-1β-induced chondrocyte was treated with A20 lentivirus activation particle, pyrrolidine dithiocarbamate (PDTC, a NF-κB inhibitor) with/without A20 siRNA. IL-6, TNF-α, and PGE2 levels were measured by ELISA, and NO production by Greiss reaction. Destabilization of the medial meniscus (DMM) surgery was used to construct the OA models, followed by injection of A20 adenovirus. MMP1 and MMP13 expression was measured by immunohistochemistry. The mRNA and protein expression were performed by qRT-PCR and western blotting, respectively.

Results

A20 was down-regulated in human OA cartilage samples, and negatively correlated with the expressions of MMP1, MMP13 and IL-1β. The IL-1β-induced chondrocyte manifested decreased A20 with increased NF-κB p65 activity. A20 overexpression suppressed the NF-κB p65 activity in IL-1β-induced chondrocyte. Furthermore, PDTC decreased IL-1β-induced chondrocyte apoptosis with the upregulated COL1A1, COL2A1, COL10A1 and ACAN, as well as the down-regulated MMP1, MMP13, COX2, iNOS, IL-6, TNF-α, NO and PGE2, which was reversed by A20 siRNA. In vivo, OA mice gained higher OARSI score and Mankin’s score, exhibited up-regulations of MMP1 and MMP13, and decreased NF-κB p65 activity, which was improved after injection of A20 adenovirus.

Conclusion

A20 was reduced in OA cartilage samples, and its overexpression, by suppressing the activity of NF-κB p65, could improve IL-1β-induced chondrocyte degradation and apoptosis in vitro, as well as mitigate the inflammation in OA mice.

Keywords:

• Osteoarthritis
• A20
• NF-κB p65
• inflammation

Disclosure statement

No potential conflict of interest was reported by the author(s).

Author contributions

Shu-Bei Cui and Tao-Xia Wang designed the study, Zhen-Wu Liu, Shu-Bei Cui and Ji-Ying Yan performed the experiment, Kai Zhang and Zhen-Wu Liu analyzed the data, Tao-Xia Wang, Kai Zhang and Ji-Ying Yan wrote the manuscript. All of them revised and approved the manuscript the manuscript.