Effects of hypoxia on the expression of inflammatory markers IL-6 and TNF-a in human normal peritoneal and adhesion fibroblasts
Effects of hypoxia on the expression of inflammatory markers IL-6 and TNF-a in human normal peritoneal and adhesion fibroblasts
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DOI：
10.3109/19396368.2012.713439
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发表时间：
2012-12-01
影响因子：
2.4
通讯作者：
Saed, Ghassan M.
中科院分区：
文献类型：
--
作者：
Ambler, Dana R.;Fletcher, Nicole M.;Saed, Ghassan M.
已知炎症参与术后粘连的形成。白细胞介素（IL）-6和肿瘤坏死因子（TNF）-α是刺激急性期反应的细胞因子，其导致全身反应，包括炎症、发热以及补体和凝血级联的激活。本研究的目的是检查这些炎症标志物的表达，在正常和缺氧条件下，在正常和粘连成纤维细胞。从同一患者的正常腹膜和粘连组织建立成纤维细胞的原代培养物，并在20%O-2或低氧2%O-2条件下培养24小时。收获细胞并分离总RNA。通过逆转录产生互补DNA，并使用IL-6和TNF-α的特异性引物进行实时RT-PCR。正常腹膜和粘连成纤维细胞均表达IL-6和TNF-α。与正常腹膜成纤维细胞相比，粘连成纤维细胞表现出显著更高水平的IL-6和TNF-α mRNA（p < 0.05）。在正常腹膜和粘连成纤维细胞中，IL-6和TNF-α mRNA水平在缺氧时均上调。正常成纤维细胞的IL-6和TNF-α mRNA水平的增加达到在粘连成纤维细胞中观察到的水平。我们的研究结果表明，缺氧通过诱导炎症标志物促进粘连表型的发展，这可能有助于术后粘连的发展。抑制炎症可能是预防和/或减少术后粘连发展的潜在治疗方法。
Inflammation is known to be involved in the postoperative adhesion development. Interleukin (IL)-6 and tumor necrosis factor (TNF)-alpha are cytokines that stimulate the acute-phase reaction, which leads to a systemic reaction including inflammation, fever, and activation of the complement and clotting cascades. The goal of this study was to examine the expression of these inflammatory markers, under normal and hypoxic conditions, in normal and adhesion fibroblasts. Primary cultures of fibroblasts were established from normal peritoneum and adhesion tissues from the same patient(s) and cultured under 20% O-2 or hypoxic 2% O-2 conditions for 24 hours. Cells were harvested and total RNA was isolated. Complimentary DNA was generated by reverse transcription and subjected to real-time RT-PCR using specific primers for IL-6 and TNF-alpha. Both normal peritoneal and adhesion fibroblasts expressed IL-6 and TNF-alpha. Adhesion fibroblasts exhibited significantly higher levels of IL-6 and TNF-alpha mRNA as compared to normal peritoneal fibroblasts (p < 0.05). Both IL-6 and TNF-alpha mRNA levels were upregulated in response to hypoxia in both normal peritoneal and adhesion fibroblasts. The increase in IL-6 and TNF-alpha mRNA levels of normal fibroblasts reached the levels observed in adhesion fibroblasts. Our results suggest that hypoxia promotes the development of the adhesion phenotype by the induction of inflammatory markers, which may contribute to the development of postoperative adhesions. The inhibition of inflammation may be a potential therapeutic approach in the prevention and/or reduction of postoperative adhesion development.