Effects of lipopolysaccharide-induced inflammation on hypoxia and inflammatory gene expression pathways of the rat testis.
Effects of lipopolysaccharide-induced inflammation on hypoxia and inflammatory gene expression pathways of the rat testis.
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DOI：
10.1186/s12610-018-0079-x
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发表时间：
2018
影响因子：
2.4
通讯作者：
London M
中科院分区：
文献类型：
--
作者：
Palladino MA;Fasano GA;Patel D;Dugan C;London M
细菌感染和睾丸炎症损害生育能力，但对睾丸炎症反应的理解是不完整的。我们感兴趣的是确定基因通路参与检测和清除感染性微生物在男性生殖道。在我们实验室以前的研究集中在睾丸的缺氧反应基因，初步实验表明，经典分类为缺氧基因的基因也在抗菌反应过程中被激活。本研究的目的是确定缺氧和炎症基因途径，有助于睾丸的抗菌保护，并考虑这些途径之间可能的串扰和相互作用。使用铜绿假单胞菌或大肠杆菌在Sprague-Dawley大鼠中诱导炎症。大肠杆菌脂多糖（LPS）。检测睾丸组织中缺氧诱导因子-1 α（HIF-1α）和核因子κ B（NF-κB）的表达水平，采用实时荧光定量PCR技术分析缺氧和炎症相关基因的表达谱。LPS组HIF-1α蛋白表达增加，而HIF-1α mRNA表达无明显变化。Western Blot分析也表明LPS处理后NF-κB和抑制性NF κ B α（IκBα）蛋白水平没有变化。5个缺氧通路基因（Angptl 4、Egr 1、Ier 3、Pai 1和Glut 1）和11个炎症通路基因（Ccl 12、Cc 13、Cd 14、Cxcl 10、Icam 1、Il 10、Il 1b、Il 6、Nfkbia、Tlr 2、Tnf）在炎症3 h后表达上调。Angptl 4、Ccl 12、Cc 13、Cd 14、Egr 1、Nfkbia、Tlr 2和Tnf在6 h时仍保持升高。6个基因（Bhlhe 40、C3、Jak 2、Nlrp 3、Slc 11 a1、Tlr 1）仅在6 h时表达上调。Tlr 5基因在3 h后表达下调，6 h后无表达。电泳迁移率变动分析结果表明LPS处理后NF-κB结合活性降低。睾丸HIF-1α在LPS诱导的炎症后上调与其他组织中HIF-1α通过NF-κB B的转录激活而上调相比，我们得出结论，睾丸中HIF-1α的上调不是通过HIF-1α mRNA的增加或通过NF-κ B依赖的机制。缺氧途径基因和参与Toll样受体（TLR）和精氨酸介导的信号传导的基因包括上调基因的主要功能类别，表明缺氧和经典炎症途径都参与睾丸的炎症反应。本文的在线版本（10.1186/s12610-018-0079-x）包含补充材料，可供授权用户使用。
Bacterial infection and inflammation of the testis impairs fertility, yet an understanding of inflammatory responses of the testis is incomplete. We are interested in identifying gene pathways involved in the detection and clearance of infectious microbes in the male reproductive tract. In previous studies in our lab focused on hypoxia-responsive genes of the testis, preliminary experiments suggested that genes classically categorized as hypoxia genes are also activated during antimicrobial responses. The purpose of this study was to identify hypoxia and inflammatory gene pathways that contribute to antimicrobial protection of the testis and to consider possible cross-talk and interactions between these pathways. Inflammation was induced in Sprague-Dawley rats using P. aeruginosa or E. coli lipopolysaccharide (LPS). Levels of hypoxia-inducible factor-1 (HIF-1α) protein and nuclear factor kappa B (NF-κB) were measured, and hypoxia and inflammatory gene expression patterns in testis were analyzed by gene expression profiling using real-time quantitative PCR arrays. In LPS-treated rats, HIF-1α protein increased with no change in Hif-1α mRNA. Western Blot analysis also demonstrated no change in NF-κB and inhibitory NFKB alpha (IκBα) protein levels following LPS treatment. Five hypoxia pathway genes (Angptl4, Egr1, Ier3, Pai1, and Glut1), and 11 inflammatory pathway genes (Ccl12, Cc13, Cd14, Cxcl10, Icam1, Il10, Il1b, Il6, Nfkbia, Tlr2, Tnf) up-regulated after 3 h of inflammation. Angptl4, Ccl12, Cc13, Cd14, Egr1, Nfkbia, Tlr2, and Tnf remained elevated at 6 h. Six genes were up-regulated at 6 h only (Bhlhe40, C3, Jak2, Nlrp3, Slc11a1, Tlr1). One gene (Tlr5) was down-regulated after 3 h and no genes at 6 h. Electrophoretic mobility shift assay results suggest a decrease in NF-κB binding activity following LPS treatment. Testicular HIF-1α is up-regulated following LPS-induced inflammation. In contrast to other tissues, in which HIF-1α is up-regulated through transcriptional activation via NF-κB, we conclude that HIF-1α in the testis is not up-regulated through an increase in Hif-1α mRNA or through NF-κB-dependent mechanisms. Hypoxia pathway genes and genes involved in Toll-like receptor (TLR) and cytokine-mediated signaling comprise major functional categories of up-regulated genes, demonstrating that both hypoxia and classic inflammatory pathways are involved in inflammatory responses of the testis. The online version of this article (10.1186/s12610-018-0079-x) contains supplementary material, which is available to authorized users.
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