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可透析白细胞提取物在骨关节炎大鼠模型中的抗炎作用:组织病理学和分子特征

Anti-inflammatory effect of dialysable leucocyte extract in a rat model of osteoarthritis: histopathological and molecular characterization.

作者信息

Acosta P, Pérez N, Pérez E, Correa B, Pérez C, Gómez C, Sánchez V, Pérez D G

机构信息

a Laboratory of Molecular Biomedicine , National School of Medicine and Homeopathy (ENMH), National Polytechnic Institute (IPN) , Mexico City , Mexico.

b Hospital Dr Victorio de la Fuente Narváez , Mexico City , Mexico.

出版信息

Scand J Rheumatol. 2016 Nov;45(6):528-535. doi: 10.3109/03009742.2016.1153140. Epub 2016 Apr 21.

Abstract

OBJECTIVES

To evaluate the effect of dialysable leucocyte extract (DLE) on pro- and anti-inflammatory profiles in a rat model of osteoarthritis (OA).

METHOD

Forty-eight male Wistar rats were divided into three groups: normal rats without treatment, OA rats treated with placebo, and OA rats treated with DLE. After treatment, the animals were killed to obtain cartilage for histological analysis and to determine the expression of pro- and anti-inflammatory cytokines by reverse transcription multiplex polymerase chain reaction (RT-MPCR) and immunohistofluorescence analyses.

RESULTS

Histological analysis revealed that OA cartilage from rats treated with DLE displayed similar characteristics to non-OA cartilage from the control group. The OA cartilage treated with placebo showed alterations in the cellular architecture and in chondrocyte cluster formation. Analysis of cytokine expression by RT-MPCR showed that OA cartilage from DLE-treated rats expressed platelet-derived growth factor (PDGF), interferon (IFN)-γ, and fibroblast growth factor (FGF)-2, similar to non-OA cartilage from the control group. However, OA cartilage from rats treated with placebo expressed interleukin (IL)-1, PDGF, and I kappa B (IκB). Confocal immunodetection of FGF-2, PDGF, and non-phosphorylated IκB showed that they were distributed in the cytoplasm of most chondrocytes in OA cartilage from DLE-treated rats whereas no nuclear factor kappa B (NF-κB) expression was observed in the nuclei. Instead, in OA cartilage from the placebo group, only weak FGF-2 staining was observed, PDGF and IκB were not detected, and NF-κB was strongly observed in both cytoplasm and nuclei.

CONCLUSIONS

Our findings suggest that DLE treatment modifies the OA process, promoting the expression of anti-inflammatory cytokines and diminishing the inflammatory effects, avoiding the nuclear translocation of NF-κB in chondrocytes.

摘要

目的

评估可透析白细胞提取物(DLE)对骨关节炎(OA)大鼠模型中促炎和抗炎谱的影响。

方法

将48只雄性Wistar大鼠分为三组:未治疗的正常大鼠、接受安慰剂治疗的OA大鼠和接受DLE治疗的OA大鼠。治疗后,处死动物以获取软骨进行组织学分析,并通过逆转录多重聚合酶链反应(RT-MPCR)和免疫组织荧光分析确定促炎和抗炎细胞因子的表达。

结果

组织学分析显示,接受DLE治疗的大鼠的OA软骨表现出与对照组非OA软骨相似的特征。接受安慰剂治疗的OA软骨在细胞结构和软骨细胞簇形成方面出现改变。通过RT-MPCR分析细胞因子表达表明,接受DLE治疗的大鼠的OA软骨表达血小板衍生生长因子(PDGF)、干扰素(IFN)-γ和成纤维细胞生长因子(FGF)-2,与对照组的非OA软骨相似。然而,接受安慰剂治疗的大鼠的OA软骨表达白细胞介素(IL)-1、PDGF和IκB。对FGF-2、PDGF和非磷酸化IκB的共聚焦免疫检测显示,它们分布在接受DLE治疗的大鼠的OA软骨中大多数软骨细胞的细胞质中,而在细胞核中未观察到核因子κB(NF-κB)表达。相反,在安慰剂组的OA软骨中,仅观察到微弱的FGF-2染色,未检测到PDGF和IκB,并且在细胞质和细胞核中均强烈观察到NF-κB。

结论

我们的研究结果表明,DLE治疗可改变OA进程,促进抗炎细胞因子的表达并减轻炎症效应,避免软骨细胞中NF-κB的核转位。

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