Vasculitis Research Unit, Department of Systemic Autoimmune Diseases, Hospital Clínic, University of Barcelona, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), , Barcelona, Catalonia, Spain.
Ann Rheum Dis. 2014 Mar;73(3):616-23. doi: 10.1136/annrheumdis-2012-202883. Epub 2013 Apr 27.
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.
Temporal artery sections from 28 patients with GCA and 22 controls were cultured in Matrigel for 5 days 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.
Histopathological features were exquisitely preserved in cultured arteries. mRNA concentrations of pro-inflammatory cytokines (particularly IL-1β and IFNγ), chemokines (CCL3/MIP-1α, CCL4/MIP-1β, CCL5/RANTES) and MMP-9 as well as IL-1β 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-1β, IL-6, TNFα, IFNγ, 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).
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.
在巨细胞动脉炎（GCA）中寻找治疗靶点受到功能系统缺乏的阻碍。我们开发了一种新的模型，由三维基质中的颞动脉培养组成，并评估了糖皮质激素治疗诱导的生物标志物变化。
来自 28 名 GCA 患者和 22 名对照者的颞动脉切片在 Matrigel 中培养 5 天，存在或不存在地塞米松。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。通过免疫测定法测量上清液中的可溶性分子。
培养的动脉中保存了极好的组织病理学特征。与对照动脉相比，培养的动脉中促炎细胞因子（特别是 IL-1β 和 IFNγ）、趋化因子（CCL3/MIP-1α、CCL4/MIP-1β、CCL5/RANTES）和 MMP-9 以及上清液中的 IL-1β 和 MMP-9 蛋白浓度的 mRNA 浓度明显更高。培养体系本身上调了对照动脉中细胞因子和血管重塑因子的表达。这最小化了患者和对照组之间的差异，但强调了观察到的变化的相关性。地塞米松下调了促炎介质（IL-1β、IL-6、TNFα、IFNγ、MMP-9、TIMP-1、CCL3 和 CXCL8）的 mRNAs，但不改变血管重塑因子（血小板衍生生长因子、MMP-2 和胶原蛋白 I 和 III）的表达。
在三维基质中的颞动脉培养过程中，患者和对照组颞动脉中基因表达的差异得以保留。糖皮质激素治疗引起的生物标志物变化与体内获得的结果非常吻合。这可能是一种合适的模型，可以探索发病机制，并进行新的治疗药物的临床前研究。
