Changes in biomarkers after therapeutic intervention in temporal arteries cultured in Matrigel: a new model for preclinical studies in giant-cell arteritis
Changes in biomarkers after therapeutic intervention in temporal arteries cultured in Matrigel: a new model for preclinical studies in giant-cell arteritis
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DOI：
10.1136/annrheumdis-2012-202883
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发表时间：
2014-03-01
影响因子：
27.4
通讯作者：
Cid, Maria C.
中科院分区：
文献类型：
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作者：
Corbera-Bellalta, Marc;Garcia-Martinez, Ana;Cid, Maria C.
研究背景巨细胞动脉炎（GCA）治疗靶点的寻找受到功能系统缺乏的阻碍。方法将28例GCA患者和22例正常对照者的颞动脉组织切片，在有或无地塞米松的基质中培养5d，观察糖皮质激素治疗后颞动脉组织中生物标志物的变化。通过实时RT-PCR评估促炎介质和血管重塑分子的组织mRNA浓度。结果培养的动脉保存了良好的组织学特征。促炎细胞因子（特别是IL-1和IFN）、趋化因子（CCL 3/MIP-1、CCL 4/MIP-1、CCL 5/RANTES）和MMP-9的mRNA浓度以及上清液中IL-1和MMP-9的蛋白浓度在来自患者的培养动脉中与对照动脉相比显著更高。培养系统本身在对照动脉中上调细胞因子和血管重塑因子的表达。这使患者和对照组之间的差异最小化，但强调了观察到的变化的相关性。地塞米松下调促炎介质（IL-1，IL-6，TNF，IFN，MMP-9，TIMP-1，CCL 3和CXCL 8）的mRNA，但不修改血管重塑因子（血小板衍生生长因子，MMP-2和胶原I和III）的表达。糖皮质激素治疗引起的生物标志物的变化与体内获得的结果令人满意。这可能是一个合适的模型，探索致病途径，并进行临床前研究与新的治疗药物。
Background Search for therapeutic targets in giant-cell arteritis (GCA) is hampered by the scarcity of functional systems. We developed a new model consisting of temporal artery culture in tri-dimensional matrix and assessed changes in biomarkers induced by glucocorticoid treatment.Methods Temporal artery sections from 28 patients with GCA and 22 controls were cultured in Matrigel for 5days in the presence or the absence of dexamethasone. Tissue mRNA concentrations of pro-inflammatory mediators and vascular remodelling molecules was assessed by real-time RT-PCR. Soluble molecules were measured in the supernatant fluid by immunoassay.Results Histopathological features were exquisitely preserved in cultured arteries. mRNA concentrations of pro-inflammatory cytokines (particularly IL-1 and IFN), chemokines (CCL3/MIP-1, CCL4/MIP-1, CCL5/RANTES) and MMP-9 as well as IL-1 and MMP-9 protein concentrations in the supernatants were significantly higher in cultured arteries from patients compared with control arteries. The culture system itself upregulated expression of cytokines and vascular remodelling factors in control arteries. This minimised differences between patients and controls but underlines the relevance of changes observed. Dexamethasone downregulated pro-inflammatory mediator (IL-1, IL-6, TNF, IFN, MMP-9, TIMP-1, CCL3 and CXCL8) mRNAs but did not modify expression of vascular remodelling factors (platelet derived growth factor, MMP-2 and collagens I and III).Conclusions Differences in gene expression in temporal arteries from patients and controls are preserved during temporal artery culture in tri-dimensional matrix. Changes in biomarkers elicited by glucocorticoid treatment satisfactorily parallel results obtained in vivo. This may be a suitable model to explore pathogenetic pathways and to perform preclinical studies with new therapeutic agents.