缺氧通过HIF-1α抑制hPDLFs内毒素耐受  

Hypoxia inhibits the Endotoxin Tolerance of h PDLFs by Upregulating HIF-1α Expression

作　　者：武曦 张纲 冯晓丹 冯小倩 谭颖徽 WU Xi;ZHANG Gang;FENG Xiao-dan;FENG Xiao-qian;TAN Ying-hui(Dept.of Stomatology,Xinqiao Hospital,Army Medical University,Chongqing,400037,China;Dept.of Oral and Maxillofacial Surgery,Xinqiao Hospital,Army Medical University,Chongqing,400037,China)

机构地区：[1]陆军军医大学新桥医院口腔科,重庆400037 [2]陆军军医大学新桥医院口腔颌面外科,重庆400037

出　　处：《现代生物医学进展》2019年第19期3611-3615,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金青年科学基金项目(81600873)

摘　　要：目的:观察缺氧对人牙周膜成纤维细胞(human Periodontal Ligament Fibroblasts,hPDLFs)内毒素耐受(Endotoxin Tolerance,ET)的影响并初步探讨其可能作用机制。方法:人牙周膜经组织块和胰蛋白酶消化,原代分离培养hPDLFs并鉴定后,给予缺氧诱导,检测其炎症因子白介素-6(Interleukin 6,IL-6)和白介素-8(Interleukin 8,IL-8)的分泌情况。进一步在常氧环境下通过慢病毒过表达增强缺氧诱导因子-1α(Hypoxia Inducible Factor 1α, HIF-1α)表达或在缺氧环境下利用特异性抑制剂YC-1 (3-(50-Hydroxymethyl-20-furyl)-1-benzylindazole)抑制hPDLFs中HIF-1α表达,分析hPDLFs炎症因子IL-6和IL-8的分泌情况的变化。结果:①缺氧时,脂多糖(Lipopolysaccharides,LPS)再次刺激hPDLFs分泌炎症因子白介素IL-6、IL-8没有明显减少(P>0.05),提示hPDLFs的ET能力受到抑制。②常氧环境下,与HIF-1α未过表达组相比,过表达组的hPDLFs受到LPS再次刺激后,其分泌炎症因子IL-6或IL-8的能力并未显著降低(P>0.05);而在缺氧环境下,hPDLFs HIF-1α表达受到抑制后,LPS再次刺激可以显著抑制hPDLFs分泌IL-6或IL-8(P<0.05)。结论:缺氧可能通过诱导HIF-1α抑制hPDLFs的ET,调节hPDLFs的免疫应答,从而加重牙周组织的免疫损伤。Objective: To observe the effect of hypoxia on the endotoxin tolerance(ET) of human periodontal ligament fibroblasts(h PDLFs), and preliminarily investigate the underlying mechanism. Methods: hPDLFs were cultured and identified after human periodontium were cut into pieces and digested with trypsin. Then to observe the effect of hypoxia and hypoxia inducible factor-1 alpha(HIF-1α)on the secretions of cytokines interleukin-6(IL-6) and IL-8 by hPDLFs, HIF-1α was overexpressed by lentivirus vector in normoxia or inhibited by specific inhibitor YC-1 in hypoxia. Results: ① When hPDLFs were stimulated by lipopolysaccharides(LPS) again, the secretions of cytokines interleukin-6(IL-6) and IL-8 were not significantly reduced compared with the initial stimulation in hypoxia(P>0.05),indicating that the activities of ET in hPDLFs were inhibited in hypoxia. ②Compared with the HIF-1α unoverexpressed group, the secretion of inflammatory factor IL-6 or IL-8 by hPDLFs was not significantly decreased in the HIF-1α overexpressed group upon LPS re-stimulation in normoxia(P>0.05), while the secretion was significantly inhibited when the expression of HIF-1α in hPDLFs was inhibited in hypoxia(P<0.05). Conclusion: Hypoxia may inhibit hPDLFs ET through inducing the expression of HIF-1α to enhance the immune injury of periodontal tissues, suggesting a new potential mechanism for hypoxia aggravating periodontitis.

关 键 词：牙周炎 牙周膜成纤维细胞 内毒素耐受 缺氧 缺氧诱导因子-1Α 

分 类 号：R-33[医药卫生] R781.4