Differential effects of Th1 versus Th2 cytokines in combination with hypoxia on HIFs and angiogenesis in RA.
Differential effects of Th1 versus Th2 cytokines in combination with hypoxia on HIFs and angiogenesis in RA.
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DOI：
10.1186/ar3934
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发表时间：
2012-08-06
影响因子：
4.9
通讯作者：
Paleolog EM
中科院分区：
文献类型：
--
作者：
Larsen H;Muz B;Khong TL;Feldmann M;Paleolog EM
缺氧和T辅助细胞1（Th 1）介导的炎症是类风湿关节炎（RA）的关键特征，并通过促进血管生成而促成疾病的发病机制。本研究的目的是探讨RA成纤维细胞样滑膜细胞（FLS）在缺氧时的血管生成基因特征，以及Th 1和Th 2细胞因子，特别是探讨缺氧和细胞因子联合作用对缺氧诱导转录因子（HIF）和血管生成的影响。人血管生成PCR阵列用于筛选暴露于缺氧（1%氧）或二甲基草酰甘氨酸（其稳定HIF）的RA FLS的cDNA。使用DNA结合测定和RNA干扰研究了缺氧诱导因子亚型在缺氧和/或细胞因子刺激的RA FLS中产生血管生成特征的参与。使用体外基于内皮的测定来测量来自缺氧处理的和/或苦参碱处理的RA FLS的条件培养基的血管生成潜力。12个血管生成基因的表达显着改变，在RA FLS暴露于缺氧，其中7个改变二甲基草酰甘氨酸，包括肝配蛋白A3（EFNA 3），血管内皮生长因子（VEGF），脂肪因子血管生成素样（ANGPTL）-4和瘦素。缺氧条件下，这4个促血管生成基因对HIF-1有不同程度的依赖性：EFNA 3>ANGPTL-4 >VEGF >leptin。Th 1细胞因子TNFα和IL-1β可诱导HIF-1的转录和活性，但不诱导HIF-2的转录和活性，且这种作用与缺氧有关。相反，Th 2细胞因子对HIF没有影响。IL-1β与缺氧协同作用，以HIF-1依赖性方式上调EFNA 3和VEGF，但尽管TNFα强烈诱导HIF-1，但其抑制脂肪因子表达，对EFNA 3的影响极小。尽管VEGF蛋白水平较高，但缺氧和TNFα处理的RA FLS的上清液诱导的内皮小管少于TNFα或缺氧处理的FLS的上清液。Th 2细胞因子IL-4通过常氧FLS强烈诱导ANGPTL-4和血管生成，并与缺氧协同诱导进一步的促血管生成活性。目前的工作表明，Th 1细胞因子与缺氧的组合是不足以诱导血管生成活性的RA FLS尽管HIF-1激活和VEGF的生产。相反，Th 2细胞因子在常氧和缺氧中诱导血管生成活性，尽管它们不能激活HIF，突出了RA中缺氧、血管生成和炎症之间的复杂关系。
Hypoxia and T-helper cell 1 (Th1) cytokine-driven inflammation are key features of rheumatoid arthritis (RA) and contribute to disease pathogenesis by promoting angiogenesis. The objective of our study was to characterise the angiogenic gene signature of RA fibroblast-like synoviocytes (FLS) in response to hypoxia, as well as Th1 and T-helper cell 2 (Th2) cytokines, and in particular to dissect out effects of combined hypoxia and cytokines on hypoxia inducible transcription factors (HIFs) and angiogenesis. Human angiogenesis PCR arrays were used to screen cDNA from RA FLS exposed to hypoxia (1% oxygen) or dimethyloxalylglycine, which stabilises HIFs. The involvement of HIF isoforms in generating the angiogenic signature of RA FLS stimulated with hypoxia and/or cytokines was investigated using a DNA-binding assay and RNA interference. The angiogenic potential of conditioned media from hypoxia-treated and/or cytokine-treated RA FLS was measured using an in vitro endothelial-based assay. Expression of 12 angiogenic genes was significantly altered in RA FLS exposed to hypoxia, and seven of these were changed by dimethyloxalylglycine, including ephrin A3 (EFNA3), vascular endothelial growth factor (VEGF), adipokines angiopoietin-like (ANGPTL)-4 and leptin. These four proangiogenic genes were dependent on HIF-1 in hypoxia to various degrees: EFNA3 >ANGPTL-4 >VEGF >leptin. The Th1 cytokines TNFα and IL-1β induced HIF-1 but not HIF-2 transcription as well as activity, and this effect was additive with hypoxia. In contrast, Th2 cytokines had no effect on HIFs. IL-1β synergised with hypoxia to upregulate EFNA3 and VEGF in a HIF-1-dependent fashion but, despite strongly inducing HIF-1, TNFα suppressed adipokine expression and had minimal effect on EFNA3. Supernatants from RA FLS subjected to hypoxia and TNFα induced fewer endothelial tubules than those from FLS subjected to TNFα or hypoxia alone, despite high VEGF protein levels. The Th2 cytokine IL-4 strongly induced ANGPTL-4 and angiogenesis by normoxic FLS and synergised with hypoxia to induce further proangiogenic activity. The present work demonstrates that Th1 cytokines in combination with hypoxia are not sufficient to induce angiogenic activity by RA FLS despite HIF-1 activation and VEGF production. In contrast, Th2 cytokines induce angiogenic activity in normoxia and hypoxia, despite their inability to activate HIFs, highlighting the complex relationships between hypoxia, angiogenesis and inflammation in RA.
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作者：
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影响因子：
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