OBJECTIVE In this work, we examine the molecular basis for capillary tube regression and identify key proregressive factors, signaling pathways, and pharmacological antagonists of this process. Approach and Results: We demonstrate that the proinflammatory mediators, IL (interleukin)-1β, TNF (tumor necrosis factor) α, and thrombin, singly and in combination, are potent regulators of capillary tube regression in vitro. These proregressive factors, when added to endothelial cell-pericyte cocultures, led to selective loss of endothelial cell-lined tube networks, with retention and proliferation of pericytes despite the marked destruction of adjacent capillary tubes. Moreover, treatment of macrophages with the TLR (toll-like receptor) agonists Pam3CSK4 and lipopolysaccharide generates conditioned media with marked proregressive activity, that is completely blocked by a combination of neutralizing antibodies directed to IL-1β and TNFα but not to other factors. The same combination of blocking antibodies, as well as the anti-inflammatory cytokine IL-10, interfere with macrophage-dependent hyaloid vasculature regression in mice suggesting that proinflammatory cytokine signaling regulates capillary regression in vivo. In addition, we identified a capillary regression signaling signature in endothelial cells downstream of these proregressive agents that is characterized by increased levels of ICAM-1 (intercellular adhesion molecule-1), phospho-p38, and phospho-MLC2 (myosin light chain-2) and decreased levels of phospho-Pak2, acetylated tubulin, phospho-cofilin, and pro-caspase3. Finally, we identified combinations of pharmacological agents (ie, FIST and FISTSB) that markedly rescue the proregressive activities of IL-1β, TNFα, and thrombin, individually and in combination. CONCLUSIONS Overall, these new studies demonstrate that the major proinflammatory mediators, IL-1β, TNFα, and thrombin, are key regulators of capillary tube regression-a critical pathological process regulating human disease.

中文翻译：

---

促炎药，IL（白介素）-1β，TNF（肿瘤坏死因子）α和凝血酶直接导致毛细血管退化。

目的在这项工作中，我们检查了毛细管回归的分子基础，并确定了该过程的关键递进因子，信号传导途径和药理拮抗剂。方法和结果：我们证明促炎性介质IL（白介素）-1β，TNF（肿瘤坏死因子）α和凝血酶单独或组合是体外毛细管降解的有效调节剂。这些进步因素，当添加到内皮细胞-周细胞共培养物中时，会导致内皮细胞内衬管网络的选择性损失，尽管相邻毛细管显着破坏，但周细胞会保留和增殖。此外，使用TLR（toll-like receptor）激动剂Pam3CSK4和脂多糖处理巨噬细胞会产生条件培养基，并具有明显的渐进活性，它被针对IL-1β和TNFα的中和抗体的组合完全阻断，但不受其他因素的阻断。阻断抗体以及抗炎细胞因子IL-10的相同组合会干扰小鼠中巨噬细胞依赖性玻璃样血管的消退，提示促炎细胞因子信号传导可调节体内的毛细血管消退。此外，我们在这些递质的下游发现了内皮细胞毛细血管递减信号，其特征是ICAM-1（细胞间粘附分子-1），磷酸化p38和磷酸化MLC2（肌球蛋白轻链2）水平升高。 ），并降低磷酸化Pak2，乙酰化微管蛋白，磷酸化纤溶蛋白和半胱天冬酶原3的水平。最后，我们确定了药理剂的组合（即，（FIST和FISTSB）分别或联合显着地拯救了IL-1β，TNFα和凝血酶的递进活性。结论总体而言，这些新研究表明，主要的促炎性介质IL-1β，TNFα和凝血酶是毛细管回归的关键调节因子，而毛细管回归是调节人类疾病的关键病理过程。