低O2高CO2对小鼠血脑屏障及水通道蛋白-4表达影响.docVIP

低O2高CO2对小鼠血脑屏障及水通道蛋白-4表达影响 　 作者：房春燕，代允义，宋兴伟，王小同 【摘要】 目的：探讨低O2高CO2对小鼠血脑屏障通透性、超微结构及水通道蛋白-4（AQP4）表达的影响。方法 ：雄性成年C57BL/6小鼠，随机分为对照组（C组）、实验组(低O2高CO2 1 d、2 d、3 d、1周、2周组)，实验组动物置于低O2高CO2舱造模。小鼠腹腔注射荧光素钠，观察不同时间点血脑屏障通透性的变化，电镜观察血脑屏障超微结构的变化，免疫组化及RT-PCR分别检测小鼠前额叶皮层及海马AQP4蛋白及mRNA表达的变化。结果 ：实验1 d组小鼠血脑屏障的通透性升高最明显，然后逐渐下降，在1周内仍显著高于对照组（Plt;0.05或Plt;0.01），2周末回落至对照组水平（P gt;0.05）。实验1 d组小鼠血脑屏障超微结构有明显改变，皮层及海马脑组织AQP4蛋白及mRNA表达显著增高（P lt;0.05）。结论 ：低O2高CO2在早期能使小鼠血脑屏障的通透性升高，可能与AQP4的表达增高有关。 【关键词】 血脑屏障；低氧；高碳酸血症；水通道蛋白-4：星形胶质细胞；小鼠 Abstract: Objective: To investigate the changes of the permeability and ultrastructure of mice’s blood-brain barrier (BBB) and expression of aquaporin-4 (AQP4) caused by hypoxic hypercapnia. Methods: The male adult C57BL/6 mice were randomly divided into six groups: the control group (C)，the hypoxic hypercapnia 1 d， 2 d， 3 d， 1 w and 2 w group. Experimental mice were exposed to normobaric hypoxic hypercapnia by placing them in a chamber. Sodium fluorescein (NaFI) was injected intraperitoneally to detect the permeability of BBB. The BBB ultrastructure was observed by electron microscope. The expression of AQP4 mRNA and protein levels were measured by Immunochemistry and PT-PCR. Results: Compared with C group， the permeability of mice’s BBB increased significantly in 1 d， 2 d， 3 d and 1 w group（Plt;0.05 or P lt;0.01），which declined gradually with the prolongation of hypoxic hypercapnic exposure and had no significant increase in 2 w group. BBB ultrastructure changes were observed by electron microscope and mRNA and protein of AQP4 were upregulated significantly in 1 d group (Plt;0.05). Conclusion: The blood-brain barrier’s permeability may increase with mice’s exposure to hypoxic hypercapnia at early stage， which may have the relationship to the upregulaion of AQP4. Key words: blood-brain barrier；hypoxia；hypercapnia；aquaporin-4；astrocyte；mouse 血脑屏障功能的调节与星形胶质细胞的足突密切相关[1]。水通道蛋白4（AQP4）是一种跨膜蛋白，在星形胶质细胞的足突上呈极化表达，是水分进出星形胶质细胞的主要通道之一。以往的研究证明，在慢性低O2高CO2条件下，实验动物认知功能发生损害[2]，其