体外培养星形细胞创伤后继发低氧二次损伤动物模型的建立
体外培养星形细胞,,创伤；星形细胞；低氧；动物模型,材料与方法,1实验动物、试剂及主要设备,2大鼠皮层星形胶质细胞的培养、纯化与鉴定,3培养
摘要: 目的 建立体外培养星形细胞牵张损伤后继发低氧二次损伤动物模型。方法 用计算机控制，以不同压力牵张损伤培养于硅胶膜培养皿上的大鼠星形细胞，并于牵张损伤后立即予以1小时低氧应激，用乳酸脱氢酶(LDH)和台盼蓝定量检测星形细胞在不同条件下和不同时相点的损伤程度。结果 不同程度的牵张压力可导致不同程度的细胞损伤，通过控制牵张压力可以调控损伤的程度；LDH与台盼蓝检测的结果明显相关；动态观察结果显示LDH的释放高峰主要在1小时之内；单纯短暂低氧应激后星形细胞LDH释放无明显升高，与正常对照组相比无显著性差异(P>0.05)。但牵张损伤后复合低氧应激后，LDH释放量明显升高，与单纯牵张损伤组相比较，有显著性差异(P<0.01)。结论 创伤后低氧可显著增加体外培养星形细胞的损伤程度，此模型可用于体外研究创伤后低氧二次打击的机制。关键词：创伤；星形细胞；低氧；动物模型
An animal model of traumatic injury with secondary hypoxia insult on cultured rats astrocytes in vitro
WANG Kewan,YANG Zhihuan,QI Songtao,et al.
(Department of Neurosurgery,Nanfang Hospital,Nanfang Medical University,Guangzhou 510515，China)
Abstract： Objective To develop a model of traumatic injury with secondary hypoxia insult on cultured rats astrocytes in vitro.Methods Rat cortical astrocytes cultured in tissue culture wells with a flexible silastic bottom were injured by stretch device.The pressure of the stretch device can be controlled and on line tested by the computer.After stretchinduced injury,the astrocytes were insulted by 1h hypoxia stress.Extent of cell injury was qualitatively assessed by lactate dehydrogenase(LDH) enzyme release and trypan blue staining.Results The diferent degree of cell injury was caused by different gas pressure.The degree of cell injury might be controlled by the different pressure.There was some significant correlation between the measured value of LDH and trypan blue staining.It was shown that the peak of LDH release is at 1h.The LDH release of astrocytes was not significantly increased after only hypoxia stress(P>0.05),but the secondary hypoxia stress after stretchinduced injury caused significant increase of LDH release of cultured astrocytes as compared with stretchinduced astrocyte(P<0.01).Conclusion Traumatic injury with secondary hypoxia insult can significantly increase the degree of injury of cultured astrocyte in vitro.The model can be used to study the mechanism of traumatic injury with secondary hypoxia insult in vitro. ......
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