NOX2 源性活性氧在胶原酶诱导的骨关节炎中的作用。

The role of NOX2-derived reactive oxygen species in collagenase-induced osteoarthritis.

机构信息

Experimental Rheumatology, Department of Rheumatology, Radboud University Medical Center, Nijmegen, The Netherlands.

Institute of Immunology, University of Münster, Germany.

出版信息

Osteoarthritis Cartilage. 2018 Dec;26(12):1722-1732. doi: 10.1016/j.joca.2018.08.014. Epub 2018 Sep 5.

DOI:10.1016/j.joca.2018.08.014

PMID:30195046

Abstract

OBJECTIVE

Synovitis in collagenase-induced osteoarthritis (CiOA) is driven by locally released S100A8/A9 proteins and enhances joint destruction. S100A8/A9 can induce reactive oxygen species (ROS) release by phagocytes in OA synovium via neutrophil cytosolic factor-1 (Ncf1)-regulated NOX2 activation. In the present study we investigated whether NOX2-derived ROS affect joint pathology during CiOA.

METHODS

CiOA was induced in knee joints of wild type (WT) and Ncf1-deficient (Ncf1**) mice. Synovial gene expression of NOX2-subunits was measured with quantitative real-time polymerase chain reaction (qRT-PCR). Joint pathology was assessed using histology and immunohistochemistry for aggrecan neo-epitope VDIPEN. Levels of inflammatory proteins were measured with Luminex or ELISA. Phagocytes in synovium, blood, bone marrow (BM) and spleen were analyzed with flow cytometry. ROS release by phagocytes was measured with a ROS detection kit.

RESULTS

CiOA induction in knee joints of WT mice caused significantly increased synovial gene expression of NOX2 subunits. On day 7 of CiOA, cartilage damage and MMP activity, as measured by VDIPEN, were comparable between WT and Ncf1** mice. Synovial thickening, synovial S100A8/A9 levels and percentages of synovial macrophages, polymorphonuclear cells (PMNs), and monocytes were not different, as were levels of inflammatory mediators in serum and phagocyte percentages in blood, BM and spleen. On day 42 of CiOA, synovitis, cartilage damage, and osteophyte formation in Ncf1** mice were unaltered when compared to WT mice. ROS detection confirmed that Ncf1** PMNs lack functional NOX2, but in vitro macrophages showed ROS production, suggesting activation of compensatory mechanisms.

CONCLUSIONS

Absence of Ncf1-mediated ROS production does not alter joint pathology in CiOA.

摘要

目的

胶原酶诱导性骨关节炎（CiOA）中的滑膜炎是由局部释放的 S100A8/A9 蛋白驱动的，会增强关节破坏。S100A8/A9 可通过中性粒细胞胞质因子-1（Ncf1）调节的 NADPH 氧化酶 2（NOX2）激活诱导 OA 滑膜中的吞噬细胞释放活性氧（ROS）。在本研究中，我们研究了 NOX2 衍生的 ROS 是否会影响 CiOA 期间的关节病理。

方法

在野生型（WT）和 Ncf1 缺陷型（Ncf1**）小鼠的膝关节中诱导 CiOA。使用实时定量聚合酶链反应（qRT-PCR）测量滑膜中 NOX2 亚基的基因表达。通过组织学和软骨寡聚基质蛋白（aggrecan）新表位 VDIPEN 的免疫组织化学评估关节病理。使用 Luminex 或 ELISA 测量炎症蛋白水平。使用流式细胞术分析滑膜、血液、骨髓（BM）和脾脏中的吞噬细胞。使用 ROS 检测试剂盒测量吞噬细胞的 ROS 释放。

结果

WT 小鼠膝关节 CiOA 诱导导致滑膜中 NOX2 亚基的基因表达显著增加。在 CiOA 的第 7 天，WT 和 Ncf1小鼠的 VDIPEN 测量的软骨损伤和 MMP 活性相当。滑膜增厚、滑膜 S100A8/A9 水平以及滑膜巨噬细胞、多形核细胞（PMN）和单核细胞的百分比在 WT 和 Ncf1小鼠之间没有差异，血清中炎症介质的水平和血液、BM 和脾脏中的吞噬细胞百分比也没有差异。在 CiOA 的第 42 天，与 WT 小鼠相比，Ncf1小鼠的滑膜炎、软骨损伤和骨赘形成没有改变。ROS 检测证实 Ncf1PMN 缺乏功能性 NOX2，但体外巨噬细胞显示 ROS 产生，表明激活了代偿机制。