Department of Anesthesia and Perioperative Care, University of California, San Francisco, CA.
Division of Pediatric Critical Care, UCSF Benioff Children's Hospitals, San Francisco, CA.
Crit Care Med. 2021 Mar 1;49(3):e315-e326. doi: 10.1097/CCM.0000000000004854.
Catecholaminergic vasopressors are the cornerstone of circulatory shock management. Nevertheless, catecholamines have problematic side effects, arousing a growing interest in noncatecholaminergic agents such as vasopressin or angiotensin-II. However, their respective effects on sepsis-associated microvascular endothelial dysfunction such as permeability or inflammation remain elusive. We investigated the role of catecholamines and other vasopressors on Toll-like receptor agonists-induced microvascular endothelial permeability and inflammation.
University research laboratory/cell research.
Human pulmonary microvascular endothelial cells from multiple donors.
Confluent monolayers of human pulmonary microvascular endothelial cells were treated with Toll-like receptor agonists (lipopolysaccharide, Poly[I:C], or tripalmitoyl-S-glyceryl cysteine) in the presence or absence of epinephrine, norepinephrine, vasopressin, and angiotensin-II. Permeability was inferred from transendothelial resistance, measured using electrical cell impedance sensing, where decreased transendothelial resistance is consistent with increased permeability. Cell-cell junction molecule expression was assessed via immunofluorescence microscopy and flow cytometry. We quantified cytokines in supernatants of Toll-like receptor agonist-treated human pulmonary microvascular endothelial cells.
Epinephrine and norepinephrine both ameliorate lipopolysaccharide, polyinosinic:polycytidylic acid, or tripalmitoyl-S-glyceryl cysteine-induced reductions in transendothelial resistance, a surrogate for endothelial permeability. In contrast, the noncatecholaminergic agents, vasopressin, and angiotensin-II did not affect Toll-like receptor agonists-induced reductions in transendothelial resistance. β1- and β2-adrenergic receptor antagonists reduced the effects of the catecholamines on transendothelial resistance, whereas α-adrenergic receptor antagonists did not. We observed that epinephrine and norepinephrine induced actin cytoskeletal rearrangement and normalized the membrane expression of proteins involved with adherens-junctions (vascular endothelial-cadherin) and tight-junctions (zona occludens-1). Despite having a substantial effect on endothelial permeability, epinephrine and norepinephrine did not affect human pulmonary microvascular endothelial cell survival or production of interleukin-8, interleukin-6, or monocyte chemoattractant protein-1 (CCL-2) induced by Toll-like receptor agonists, suggesting that these functions are regulated separately from permeability.
Our findings demonstrate that treatment with epinephrine or norepinephrine strongly reduces endothelial permeability induced by agonists of multiple Toll-like receptors (Toll-like receptor-2, Toll-like receptor-3, Toll-like receptor-4) in vitro. Our studies suggest that both β1- and β2-adrenergic receptors mediate the stabilizing effects of epinephrine and norepinephrine on the endothelial barrier.
儿茶酚胺类血管加压素是休克治疗的基石。然而，儿茶酚胺有其问题的副作用，这引起了人们对非儿茶酚胺类药物如血管加压素或血管紧张素-II 的日益关注。然而，它们对脓毒症相关的微血管内皮功能障碍（如通透性或炎症）的各自影响仍不清楚。我们研究了儿茶酚胺和其他血管加压素对 Toll 样受体激动剂诱导的微血管内皮通透性和炎症的作用。
大学研究实验室/细胞研究。
来自多个供体的人肺微血管内皮细胞。
在存在或不存在肾上腺素、去甲肾上腺素、血管加压素和血管紧张素-II 的情况下，用 Toll 样受体激动剂（脂多糖、聚[I：C]或三棕榈酰-S-甘油半胱氨酸）处理人肺微血管内皮细胞的汇合单层。通过使用电细胞阻抗传感测量跨内皮电阻来推断通透性，跨内皮电阻的降低与通透性的增加一致。通过免疫荧光显微镜和流式细胞术评估细胞-细胞连接分子的表达。我们定量了 Toll 样受体激动剂处理的人肺微血管内皮细胞上清液中的细胞因子。
肾上腺素和去甲肾上腺素均可改善脂多糖、聚肌苷酸：聚胞苷酸或三棕榈酰-S-甘油半胱氨酸诱导的跨内皮电阻降低，跨内皮电阻降低是内皮通透性的替代物。相比之下，非儿茶酚胺类药物血管加压素和血管紧张素-II 并不影响 Toll 样受体激动剂诱导的跨内皮电阻降低。β1-和β2-肾上腺素能受体拮抗剂降低了儿茶酚胺对跨内皮电阻的作用，而α-肾上腺素能受体拮抗剂则没有。我们观察到肾上腺素和去甲肾上腺素诱导了肌动蛋白细胞骨架的重排，并使参与黏附连接（血管内皮钙粘蛋白）和紧密连接（闭合蛋白-1）的蛋白的膜表达正常化。尽管对内皮通透性有很大影响，但肾上腺素和去甲肾上腺素并不影响 Toll 样受体激动剂诱导的人肺微血管内皮细胞的存活或白细胞介素-8、白细胞介素-6 或单核细胞趋化蛋白-1（CCL-2）的产生，这表明这些功能与通透性分开调节。
我们的发现表明，肾上腺素或去甲肾上腺素的治疗可强烈降低体外多种 Toll 样受体（Toll 样受体-2、Toll 样受体-3、Toll 样受体-4）激动剂诱导的内皮通透性。我们的研究表明，β1-和β2-肾上腺素能受体均介导肾上腺素和去甲肾上腺素对内皮屏障的稳定作用。
