中文摘要：

背景脑膜上皮细胞（MECs）是构成脑脊液-视神经屏障的主要细胞成分。脑脊液-视神经屏障的损害可能会导致脑脊液组分的失衡，使视神经受到各种致病因素的攻击。目前对于MECs在视神经疾病中的病理作用研究甚少，机制尚不明确。目的探讨缺氧条件下人MECs的功能变化，为视神经疾病发病机制的研究提供新的线索。方法将培养的人MECs株分别制备成密度为2．5×10^3个／孔、5．0×10^3个／孔和1×10^4个／孔的细胞悬液，各取100μl分别接种于96孔板中，分别在常规培养基及体积分数21％O2（常氧组）和1％O2（缺氧组）环境下孵育2d，采用MTS法测定和比较不同氧环境下人MECs的吸光度（A490）值；采用CASY1法检测和比较不同氧环境下细胞体积及直径的变化；采用光度计测定MECs暴露不同氧环境下1d、2d后线粒体产生ATP量的变化；采用免疫荧光技术测定不同氧环境下细胞内细胞色素C的表达和定位。结果缺氧组2．5×10^3个／孔、5．0×10^3个／孔、1×10^4个／孔细胞密度组MECs的增生值（A490）分别为0．399±0．009、0．393±0．009和0．496±0．026，分别较相应的常氧组的0．424±0．131、0．413±0．111和0．537±0．021明显下降，差异均有统计学意义（t=3．777，P=0．004；t=3．251，P=0．009；t=3．037，P=0．013）。与常氧组细胞比较，缺氧组细胞的直径和体积均明显增加[（20．970±0．127）μm vs．（21．198±0．048）μm，t=-3．762，P=0．006；（5805±73）fl vs．（6026±106）fl，t=-4．124，P=0．002）]。缺氧组和缺氧＋底物组细胞分别培养2d后，细胞中ATP产生量分别为（0．900±0．225）mmol／（L·g）、（0．952±0．075）mmol／（L·g），均明显低于常氧组的（1．389±0．145）mmol／（L·g）和常氧＋底物组的（1．401±0．122）mmol／（L·g），差异均有统计学意义（P=0．001、0．002、0．001）。常氧组细胞中细?

英文摘要：

Background BackgroundMeniugothelial cells （MECs） are major cell type in the meningeal sheath around optic nerve,which form a fluid barrier between optic nerve and the cerebral spinal fluid. The impairment of the cerebral fluid-optic nerve barrier probably affects the balance of cerebral fluid components. Currently, the investigation on the role of MECs in neuropathy is less performed. Objective This study attempted to explore hypoxia-induced function changes of MECs,and to shed a new clus for the future research of optic nerve disorders. Methods Human MECs strains were cultured in vitro and cell suspension was prepared with the cell densities of 2.5 ×10^3/hole,5.0× 10^3/hole and 1 × 10^4/hole, respectively. The suspensions of 100 μl were separately collected to incubate in 96-well plates and cultivated for 2 days in 21% O2（ normoxia group） or 1% O2（ hypoxia group）. MTS was used to detect and compare the proliferative value （A490 ） of MECs between the normoxia group and the hypoxia group. The changes of MECs diameter and volume were measured by CASY1 assay. ATP product in the cells after MECs exposed to different oxygen environments with or without substrate （ 100 mmol/L pyruvate and 100 mmol/L malate） for 1,2 days were assayed by Luminometer method. The expression and distribution of cytochrome C in the ceils of the normoxia group and the hypoxia group were determined by immunofluorescenee. Results A490 of MECs in the 2.5×10^3/hole, 5. 0× 10^3/hole and 1×10^4/hole were 0. 399 ±0. 009,0. 393 ±0. 009 and 0. 496±0. 026 in the hypoxia group,which were lower than 0. 424±0. 131,0. 413±0. 111 and 0. 537±0. 021 in the normoxia group （ t = 3. 777,P=0. 004; t=3. 251 ,P=0. 009; t=3. 037,P=0. 013）. Compared with the normoxia group,the diameter and volume were significantly increased in the hypoxia group （ [ 20. 970 ±0. 127 ] μm vs. [ 21. 198 ± 0. 048 ] μm, t = -3.762,P=0.006; [5805±73] fl vs. [6026±106] fl,t=-4.124,P=0. 002）.ATP products were （0.900±0.225） mmol/（L · g）