Caffeine Treatment Promotes Differentiation and Maturation of Hypoxic Oligodendrocytes via Counterbalancing Adenosine 1 Adenosine Receptor-Induced Calcium Overload (Retracted Article)
Caffeine Treatment Promotes Differentiation and Maturation of Hypoxic Oligodendrocytes via Counterbalancing Adenosine 1 Adenosine Receptor-Induced Calcium Overload (Retracted Article)
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DOI：
10.12659/msm.915147
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发表时间：
2019-03-06
影响因子：
3.1
通讯作者：
Li, Hongli
中科院分区：
文献类型：
--
作者：
Cao, Ting;Ma, Teng;Li, Hongli
背景：我们旨在探讨腺苷1腺苷受体（A1AR）在缺氧诱导的少突胶质细胞（OLs）分化不良中的作用，以及咖啡因治疗缺氧损伤的潜在机制。材料/方法：采用实时聚合酶链反应（RT-PCR）技术检测培养的缺氧OLs在咖啡因处理或不处理的情况下AR表达的变化。然后，通过共聚焦Fluo-3成像检测细胞内Ca2+浓度（[Ca2+]）的变化。western blot和免疫荧光法检测髓磷脂相关蛋白的表达变化。结果：缺氧后3 h， A1AR表达显著上调，少突胶质细胞转录因子（Olig2）表达显著降低。此外，缺氧刺激或100 μ M腺苷均可诱导培养的OLs静息[Ca2+]明显升高。然而，dppcpx （A1AR选择性拮抗剂）或咖啡因预处理消除了[Ca2+]的增加，随后的高剂量腺苷诱导了发展中的OLs中的Ca2+活性。此外，咖啡因或DPCPX在缺氧刺激后提高了MBP和CNPase蛋白的表达，导致OLs形态成熟。结论：咖啡因对新生儿缺氧损伤有保护作用。它可以防止Ca2+过载损伤，在缺氧发育的OLs中保持Ca2+稳态，并通过抑制A1AR促进髓磷脂相关蛋白的最佳表达。本研究也为咖啡因在新生儿缺氧早期治疗中的临床应用提供了实验依据，并突出了A1AR在抗缺氧药物发现中的潜在意义。
Background: We aimed to explore the involvement of adenosine 1 adenosine receptor (A1AR) in hypoxia-induced poor differentiation of oligodendrocytes (OLs), and the underlying mechanism of caffeine treatment in hypoxic injuries.Material/Methods: Real-time polymerase chain reaction (RT-PCR) was used to assess the alterations of AR expression in cultured hypoxic OLs with or without caffeine treatment. Then, intracellular alterations of Ca2+ concentrations ([Ca2+]) were detected by confocal Fluo-3 imaging. The subsequent changes of myelin related protein expression were determined by western blot and immunofluorescence.Results: Three hours after hypoxia, significantly upregulated expression of A1AR was observed, accompanied with significantly decreased expression of oligodendrocyte transcription factor (Olig2). In addition, either hypoxia stimulation or 100 mu M adenosine induced apparent elevation of resting [Ca2+] in cultured OLs. However, pretreatment with DPCPX (A1AR selective antagonist) or caffeine abolished the [Ca2+] increase, and the subsequent adenosine of high dose induced Ca2+ activity in developing OLs. Furthermore, caffeine or DPCPX improved the expression MBP and CNPase proteins after hypoxia stimulation, which resulted in the morphological maturation of OLs.Conclusions: Caffeine treatment exerted protective effects on neonatal hypoxia injuries. It prevented Ca2+ overload injury, kept Ca2+ homeostasis in hypoxic developing OLs, and facilitated optimal expression of myelin related proteins by inhibiting A1AR in vitro. This study also provided experimental evidence for clinical application of caffeine in early treatment of neonatal hypoxia, and highlighted the potential significance of A1AR in anti-hypoxic drug discovery.