Ethanol consumption during pregnancy or lactation period can induce permanent damage to the development of the central nervous system (CNS), resulting in fetal alcohol spectrum disorders (FASD). CNS development depends on proper neural cells and blood vessel (BV) development and blood-brain barrier (BBB) establishment; however, little is known about how ethanol affects these events. Here, we investigated the impact of ethanol exposure to endothelial cells (ECs) function and to ECs interaction with astrocytes in the context of BBB establishment. Cerebral cortex of newborn mice exposed in utero to ethanol (FASD model) presented a hypervascularized phenotype, revealed by augmented vessel density, length, and branch points. Further, aberrant distribution of the tight junction ZO-1 protein along BVs and increased rates of perivascular astrocytic endfeet around BVs were observed. In vitro exposure of human brain microcapillary ECs (HBMEC) to ethanol significantly disrupted ZO-1 distribution, decreased Claudin-5 and GLUT-1 expression and impaired glucose uptake, and increased nitric oxide secretion. These events were accompanied by upregulation of angiogenesis-related secreted proteins by ECs in response to ethanol exposure. Treatment of cortical astrocytes with conditioned medium (CM) from ethanol exposed ECs, upregulated astrocyte’s expression of GFAP, Cx43, and Lipocalin-2 genes, as well as the pro-inflammatory genes, IL-1beta, IL-6, and TNF-alpha, which was accompanied by NF-kappa B protein nuclear accumulation. Our findings suggest that ethanol triggers a dysfunctional phenotype in brain ECs, leading to impairment of cortical vascular network formation, and promotes ECs-induced astrocyte dysfunction, which could dramatically affect BBB establishment in the developing brain.

怀孕或哺乳期间饮酒会导致中枢神经系统 (CNS) 发育的永久性损害，导致胎儿酒精谱系障碍 (FASD)。中枢神经系统发育依赖于适当的神经细胞和血管（BV）发育以及血脑屏障（BBB）的建立；然而，人们对乙醇如何影响这些事件知之甚少。在这里，我们研究了乙醇暴露对内皮细胞 (ECs) 功能的影响以及在 BBB 建立的背景下对 ECs 与星形胶质细胞相互作用的影响。子宫内暴露于乙醇（FASD 模型）的新生小鼠的大脑皮层呈现出血管丰富的表型，通过增加的血管密度、长度和分支点来揭示。更远，观察到紧密连接 ZO-1 蛋白沿 BVs 的异常分布和 BVs 周围血管周围星形胶质细胞末端的发生率增加。人脑微毛细血管内皮细胞 (HBMEC) 体外暴露于乙醇会显着破坏 ZO-1 分布，降低 Claudin-5 和 GLUT-1 表达，降低葡萄糖摄取，并增加一氧化氮分泌。这些事件伴随着 ECs 响应乙醇暴露而上调血管生成相关分泌蛋白。用来自乙醇暴露的 ECs 的条件培养基 (CM) 处理皮质星形胶质细胞，上调星形胶质细胞的 GFAP、Cx43 和 Lipocalin-2 基因以及促炎基因 IL-1beta、IL-6 和 TNF-alpha 的表达, 伴随着 NF-kappa B 蛋白核积累。