Changes in membrane properties during energy depletion-induced cell injury studied with fluorescence microscopy.
Changes in membrane properties during energy depletion-induced cell injury studied with fluorescence microscopy.
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用荧光显微镜研究能量消耗引起的细胞损伤期间膜特性的变化。
DOI：
10.1016/s0006-3495(96)79243-3
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发表时间：
1996
影响因子：
3.4
通讯作者：
B. Taylor
中科院分区：
文献类型：
--
作者：
Yan;F. Sun;Donald M. Tong;B. Taylor
本文用稳态荧光各向异性（r）和光漂白后荧光再分布（FRAP）技术研究了ATP耗竭诱导的人星形细胞瘤细胞（UC-11 MG）损伤过程中膜结构特性的变化。建立了一种新的测量r的方法，该方法提供了同时监测计算r所需的垂直和水平偏振荧光发射的独特优势。在本研究中，用1-（4-三甲基铵苯基）-6-苯基-1，3，5-己三烯对甲苯磺酸盐标记的星形细胞瘤细胞中的r被证明保持稳定长达90分钟。然而，当细胞用75 μ M碘乙酸（IAA），一种代谢抑制剂，诱导细胞ATP的快速耗尽，r不断下降，表明膜脂质秩序和双层结构的扰动减少。用亲脂性抗氧化剂（如替拉扎德或棉酚）对细胞进行预处理，可以防止r值的下降。Tirilazad本身引起r的显著增加，表明tirilazad插入膜双层并深刻地增加未损伤细胞中的脂质秩序。然而，棉酚并没有表现出这种性质。进一步用FRAP研究这些现象证实了上述结果，并表明在ATP诱导的细胞损伤过程中，膜流动性增加，而其结构变得不那么刚性。此外，亲脂性抗氧化剂防止IAA诱导的膜结构畸变。实验结果表明，不同的细胞保护作用机制可能存在替拉唑和抗氧化剂棉酚。棉酚似乎完全是因为其抗氧化作用而防止或延迟观察到的细胞损伤，而替拉扎德的保护不仅通过其抗氧化活性介导，而且还通过其增加细胞膜脂质顺序的能力介导。
The changes in membrane structural properties occurring during the process of ATP depletion-induced cell injury in adherent human astrocytoma cells (UC-11 MG) were studied with two epifluorescence techniques: 1) steady-state fluorescence anisotropy (r) to examine microstructural changes in the membrane phospholipids and 2) fluorescence redistribution after photobleaching (FRAP) to examine membrane fluidity changes. A new method for r measurement was established that provides the unique advantage of simultaneously monitoring both vertical and horizontal polarized fluorescence emissions needed for the calculation of r. In this study, r in the astrocytoma cells labeled with 1-(4-trimethylammonium phenyl)-6-phenyl-1,3,5-hexatriene p-toluenesulfonate was shown to remain stable for up to 90min. However, when the cells were treated with 75 microM iodoacetic acid (IAA), a metabolic inhibitor that induces rapid depletion of cellular ATP, r continually decreased, indicating a decrease in membrane lipid order and perturbation of the bilayer structure. This decrease in r could be prevented by the pretreatment of cells with lipophilic antioxidants such as tirilazad or gossypol. Tirilazad itself caused a significant increase in r, suggesting that tirilazad intercalates into the membrane bilayer and profoundly increases the lipid order in uninjured cells. Gossypol, however, did not exhibit this property. Further investigations into these phenomena with FRAP confirmed the r results and indicated that membrane fluidity increased while its structure became less rigid during the process of ATP-induced cell injury. In addition, lipophilic antioxidants prevented the membrane structural aberrations induced by IAA. Experimental results suggest that different mechanisms of cytoprotective action may exist for tirilazad and the antioxidant gossypol. Gossypol appears to prevent or delay the observed cell injury entirely because of its antioxidant action, whereas tirilazad's protection is mediated not only via its antioxidant activity, but also by its ability to increase cell membrane lipid order.
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影响因子：
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期刊：
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发表时间：
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