锌指蛋白 A20 通过影响 NF-κB p65 的活性来调节骨关节炎的发生和发展。

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

机构信息

The First Department of Orthopedics, Handan Central Hospital, Handan, China.

Department of Nephrology, Affiliated Hospital of Hebei University of Technology, Handan, China.

出版信息

Immunopharmacol Immunotoxicol. 2021 Dec;43(6):713-723. doi: 10.1080/08923973.2021.1970764. Epub 2021 Aug 31.

DOI:10.1080/08923973.2021.1970764

PMID:34463587

Abstract

OBJECTIVE

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

METHODS

and 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

was down-regulated in human OA cartilage samples, and negatively correlated with the expressions of and . 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. , 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 , as well as mitigate the inflammation in OA mice.

摘要

目的

通过调节 NF-κB p65 来研究锌指蛋白 A20 在骨关节炎（OA）中的作用。

方法

通过 qRT-PCR 检测人 OA 软骨样本中和的表达。用 A20 慢病毒激活粒子、吡咯烷二硫代氨基甲酸盐（PDTC，NF-κB 抑制剂）处理 IL-1β 诱导的软骨细胞，并用/不用 A20 siRNA。通过 ELISA 测定 IL-6、TNF-α 和 PGE2 水平，通过 Greiss 反应测定 NO 产生。采用内侧半月板不稳定（DMM）手术构建 OA 模型，然后注射 A20 腺病毒。通过免疫组化测定 MMP1 和 MMP13 的表达。通过 qRT-PCR 和 Western blot 分别进行 mRNA 和蛋白表达检测。

结果

在人 OA 软骨样本中下调，与和的表达呈负相关。IL-1β 诱导的软骨细胞表现出 A20 减少，NF-κB p65 活性增加。A20 过表达抑制了 IL-1β 诱导的软骨细胞中 NF-κB p65 的活性。此外，PDTC 降低了 IL-1β 诱导的软骨细胞凋亡，同时上调了 COL1A1、COL2A1、COL10A1 和 ACAN，下调了 MMP1、MMP13、COX2、iNOS、IL-6、TNF-α、NO 和 PGE2，而 A20 siRNA 则逆转了这一结果。在 OA 小鼠中，A20 腺病毒注射后，OARSI 评分和 Mankin 评分升高，MMP1 和 MMP13 上调，NF-κB p65 活性降低。