Lin Shishui, Qiu Meiguang, Chen Jianting
Department of Spinal Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, China.
Cell Physiol Biochem. 2015;35(6):2213-22. doi: 10.1159/000374026. Epub 2015 Apr 8.
BACKGROUND/AIMS: Osteoclasts (OC) originate from monocytes/macrophages and play a critical role in the development of ankylosing spondylitis (AS). Receptor activator of NF-kB ligand (RANKL) is critical for the differentiation and maturation of OC from monocytes/macrophages, the underlying mechanisms of which processes have not been completely elucidated. Interleukin 4 (IL-4) attenuates the pathogenesis of AS via ill-defined mechanisms.
We used a proteoglycan-induced arthritis (PGIA) model in Balb/c mice to study AS in humans. We injected IL-4 into the articular cavity and evaluated its effects on PGIA by incidence of arthritis, clinical and pathological arthritis severity and PET tracer uptake. We isolated and analyzed the number and polarization of macrophages in the articular cavity before and after IL-4 treatment. We analyzed RANKL levels in macrophage subtypes. We then isolated bone-marrow derived macrophages and treated them with IL-4 in vitro, with or without histone deacetylase inhibitors trichostatin A (TSA), and then analyzed the polarization of cultured macrophages before and after IL-4 treatment and RANKL levels in macrophage subtypes.
IL-4 treatment decreased the incidence and severity of arthritis in a mouse AS model, and polarized macrophages from a classical M1 subtype to an M2 subtype in vivo and in vitro. RANKL was predominantly produced by M1, but not by M2 macrophages. IL-4-mediated inhibition of RANKL in macrophages was abolished by TSA.
Our data suggest that the therapeutic effects of IL-4 on AS may result from a M1-to-M2 macrophage polarization and its inhibition of RANKL expression on macrophages, possibly through enhanced histone deacetylation.
背景/目的:破骨细胞(OC)起源于单核细胞/巨噬细胞,在强直性脊柱炎(AS)的发展中起关键作用。核因子κB受体活化因子配体(RANKL)对于单核细胞/巨噬细胞向OC的分化和成熟至关重要,但其相关过程的潜在机制尚未完全阐明。白细胞介素4(IL-4)通过不明机制减轻AS的发病机制。
我们在Balb/c小鼠中使用蛋白聚糖诱导的关节炎(PGIA)模型来研究人类AS。我们将IL-4注入关节腔,并通过关节炎发病率、临床和病理关节炎严重程度以及PET示踪剂摄取来评估其对PGIA的影响。我们分离并分析了IL-4治疗前后关节腔中巨噬细胞的数量和极化情况。我们分析了巨噬细胞亚型中的RANKL水平。然后我们分离骨髓来源的巨噬细胞,并在体外使用或不使用组蛋白去乙酰化酶抑制剂曲古抑菌素A(TSA)对其进行IL-4处理,接着分析IL-4处理前后培养巨噬细胞的极化情况以及巨噬细胞亚型中的RANKL水平。
IL-4治疗降低了小鼠AS模型中关节炎的发病率和严重程度,并在体内和体外将巨噬细胞从经典的M1亚型极化为M2亚型。RANKL主要由M1巨噬细胞产生,而不是M2巨噬细胞。TSA消除了IL-4介导的对巨噬细胞中RANKL的抑制作用。
我们的数据表明,IL-4对AS的治疗作用可能源于M1向M2巨噬细胞极化及其对巨噬细胞上RANKL表达的抑制,可能是通过增强组蛋白去乙酰化作用实现的。