Gene transfer of high-mobility group box 1 box-A domain in a rat acute liver failure model
Gene transfer of high-mobility group box 1 box-A domain in a rat acute liver failure model
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DOI：
10.1016/j.jss.2014.11.022
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发表时间：
2015-04-01
影响因子：
2.2
通讯作者：
Kitagawa, Yuko
中科院分区：
文献类型：
--
作者：
Tanaka, Masayuki;Shinoda, Masahiro;Kitagawa, Yuko
背景：高迁移率族蛋白1（HMGB1）最近被确定为各种急慢性炎症的重要介质。 HMGB1 基因的 box-A 结构域编码的蛋白质已知可作为 HMGB1 的竞争性抑制剂。在本研究中，我们研究了box-A基因转移是否会导致大鼠体内box-A蛋白的产生，并使用急性肝衰竭（ALF）模型评估体内治疗效果。 材料和方法：构建了三种类型的腺病毒载体（一种野生型和两种突变型），然后根据HeLa细胞的分泌情况选择突变型载体。对分泌的蛋白质进行体外肿瘤坏死因子(TNF)产生抑制试验。该载体通过健康 Wistar 大鼠的门静脉注射，以确认肝脏中 Box-A 蛋白的产生。然后将该载体通过门静脉注射到患有 ALF 的大鼠中。结果：Western blot 分析显示，用一种突变载体转染的 HeLa 细胞中 box-A 蛋白的表达增强。转染该载体的 HeLa 细胞的培养上清液抑制巨噬细胞产生 TNF-α。转染后72小时，在转染的肝脏中证实了box-A蛋白的表达。转染的大鼠表现出肝酶、血浆 HMGB1 和肝脏 TNF-α 信使 RNA 水平降低，组织学结果和生存率显着改善。结论：HMGB1 box-A 基因转移导致肝脏中 box-A 蛋白的产生，似乎对大鼠 ALF 具有有益作用。 (C) 2015 Elsevier Inc. 保留所有权利。
Background: High-mobility group box 1 (HMGB1) has recently been identified as an important mediator of various kinds of acute and chronic inflammation. The protein encoded by the box-A domain of the HMGB1 gene is known to act as a competitive inhibitor of HMGB1. In this study, we investigated whether box-A gene transfer results in box-A protein production in rats and assessed therapeutic efficacy in vivo using an acute liver failure (ALF) model.Materials and methods: Three types of adenovirus vectors were constructed-a wild type and two mutants-and a mutant vector was then selected based on the secretion from HeLa cells. The secreted protein was subjected to a tumor necrosis factor (TNF) production inhibition test in vitro. The vector was injected via the portal vein in healthy Wistar rats to confirm box-A protein production in the liver. The vector was then injected via the portal vein in rats with ALF.Results: Western blot analysis showed enhanced expression of box-A protein in HeLa cells transfected with one of the mutant vectors. The culture supernatant from HeLa cells transfected with the vector inhibited TNF-alpha production from macrophages. Expression of box-A protein was confirmed in the transfected liver at 72 h after transfection. Transfected rats showed decreased hepatic enzymes, plasma HMGB1, and hepatic TNF-alpha messenger RNA levels, and histologic findings and survival were significantly improved.Conclusions: HMGB1 box-A gene transfer results in box-A protein production in the liver and appears to have a beneficial effect on ALF in rats. (C) 2015 Elsevier Inc. All rights reserved.