C2-ceramide influences alveolar epithelial barrier function by downregulating Zo-1, occludin and claudin-4 expression

IF 2.8 4区医学 Q2 TOXICOLOGY Toxicology Mechanisms and Methods Pub Date : 2017-01-24 DOI:10.1080/15376516.2017.1278812

Jin Yang, Ying Wang, Hui Liu, Ji-rui Bi, Youjin Lu

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引用次数: 22

Abstract

Abstract Noncardiogenic lung edema is a key factor affecting the prognosis of acute lung injury (ALI). Previous studies have been focused on regulatory roles of ceramide on lung vascular endothelial barrier functions and had already identified ceramide as mediator involved in the formation of lung edema. However, the effects of ceramide on lung epithelial barrier were still unknown. This study aimed to investigate the effects of ceramide on the barrier function of alveolar epithelial cells. Primary mouse alveolar type II epithelial cells (AECII) were grown on Transwell polyester membranes to construct monolayer, and stimulated with different concentrations of ceramide. Transepithelial resistance (TER) was measured to assess the epithelial cell permeability. Western blotting and real-time quantitative polymerase chain reaction were used to detect the mRNA and protein levels of tight junction, respectively. After incubation with different concentrations of c2-ceramide, TER of AECII monolayer decreased significantly in a dose-dependent manner. Moreover, expressions of ZO-1, occludin and claudin-4 were significantly reduced by c2-ceramide in the study. This study demonstrated that ceramide could increase alveolar epithelial cell monolayer permeability by downregulation of tight junction proteins. Therefore, modulation of ceramide expression may serve as a new therapeutic approach to treat acute lung injury.

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C2神经酰胺通过下调Zo-1、occludin和claudin-4的表达影响肺泡上皮屏障功能

摘要非心源性肺水肿是影响急性肺损伤（ALI）预后的关键因素。先前的研究集中在神经酰胺对肺血管内皮屏障功能的调节作用上，并且已经确定神经酰胺是参与肺水肿形成的介质。然而，神经酰胺对肺上皮屏障的影响尚不清楚。本研究旨在探讨神经酰胺对肺泡上皮细胞屏障功能的影响。原代小鼠肺泡II型上皮细胞（AECII）在Transwell聚酯膜上生长以构建单层，并用不同浓度的神经酰胺刺激。通过测量跨上皮电阻（TER）来评估上皮细胞的通透性。采用蛋白质印迹和实时定量聚合酶链反应分别检测紧密连接的mRNA和蛋白质水平。在与不同浓度的c2神经酰胺孵育后，AECII单层的TER以剂量依赖的方式显著降低。此外，在研究中，c2神经酰胺显著降低了ZO-1、occludin和claudin-4的表达。本研究表明，神经酰胺可以通过下调紧密连接蛋白来增加肺泡上皮细胞单层的通透性。因此，调节神经酰胺的表达可能成为治疗急性肺损伤的一种新的治疗方法。

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来源期刊

Toxicology Mechanisms and Methods TOXICOLOGY-

自引率

3.10%

发文量

期刊介绍： Toxicology Mechanisms and Methods is a peer-reviewed journal whose aim is twofold. Firstly, the journal contains original research on subjects dealing with the mechanisms by which foreign chemicals cause toxic tissue injury. Chemical substances of interest include industrial compounds, environmental pollutants, hazardous wastes, drugs, pesticides, and chemical warfare agents. The scope of the journal spans from molecular and cellular mechanisms of action to the consideration of mechanistic evidence in establishing regulatory policy. Secondly, the journal addresses aspects of the development, validation, and application of new and existing laboratory methods, techniques, and equipment. A variety of research methods are discussed, including: In vivo studies with standard and alternative species In vitro studies and alternative methodologies Molecular, biochemical, and cellular techniques Pharmacokinetics and pharmacodynamics Mathematical modeling and computer programs Forensic analyses Risk assessment Data collection and analysis.