HMGB1 B box increases the permeability of Caco-2 enterocytic monolayers and impairs intestinal barrier function in mice
HMGB1 B box increases the permeability of Caco-2 enterocytic monolayers and impairs intestinal barrier function in mice
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DOI：
10.1053/gast.2002.35391
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发表时间：
2002-09-01
期刊：
影响因子：
29.4
通讯作者：
Fink, MP
中科院分区：
文献类型：
--
作者：
Sappington, PL;Yang, R;Fink, MP
背景和目标：高迁移率族（HMG）B1是一种非组蛋白核蛋白，最近被确定为脂多糖诱导的小鼠致死性的迟效介质。HMGB1的促炎作用定位于分子的一个区域，称为B盒。研究方法：为了确定HMGB：1或B盒是否能够引起肠屏障功能的紊乱，我们将培养的Caco-2人肠细胞单层与重组人HMGB 1或由B盒结构域组成的74个残基截短形式的蛋白质孵育。结果如下：HMGB1和B box均以时间和剂量依赖性方式增加Caco-2单层对异硫氰酸荧光素标记的葡聚糖（17134）的渗透性。去除重组蛋白后，渗透性的增加是可逆的。Caco-2细胞暴露于B盒导致诱导型一氧化氮合酶信使RNA表达增加和NO产生增加。当我们使用各种药理学策略来抑制NO产生或过氧化亚硝酸盐（ONOO-）时，我们消除了B盒诱导的高通透性。给野生型小鼠施用B box增加了回肠粘膜对FD 4的渗透性和细菌向肠系膜淋巴结的移位。在诱导型一氧化氮合酶基因敲除小鼠中未观察到这些效应。结论：这些数据支持HMGB1和B box能够通过依赖于NO和ONOO-形成的机制引起肠道屏障功能改变的观点。
Background & Aims: High mobility group (HMG) B1 is a nonhistone nuclear protein that was recently identified as a late-acting mediator of lipopolysaccharide-induced lethality in mice. The proinflammatory actions of HMGB1 have been localized to a region of the molecule called the B box. Methods: To determine whether HMGB:l or B box are capable of causing derangements in intestinal barrier function, we incubated cultured Caco-2 human enterocytic monolayers with recombinant human HMGB1 or a 74-residue truncated form of the protein consisting of the B box domain. Results: Both HMGB1 and B box increased the permeability of Caco-2 monolayers to fluorescein Isothiocyanate-labeled dextran (17134) in a time- and dose-dependent fashion. The increase in permeability was reversible following removal of the recombinant protein. Exposure of Caco-2 cells to B box resulted in increased expression of inducible nitric oxide synthase messenger RNA and increased production of NO. When we used various pharmacologic strategies to inhibit NO production or scavenge NO or peroxynitrite (ONOO-), we abrogated B box-induced hyperpermeability. Administration of B box to wild-type mice increased both ileal mucosal permeability to FD4 and bacterial translocation to mesenteric lymph nodes. These effects were not observed in inducible nitric oxide synthase knockout mice. Conclusions: These data support the view that HMGB1 and B box are capable of causing alterations in gut barrier function via a mechanism that depends on the formation of NO and ONOO-.