米诺环素对压力作用下体外培养视网膜神经细胞凋亡的抑制  

Inhibition effect of minocyclinye on pressure-induced apoptosis of retinal neuron cells cultured in vitro

作　　者：刘清[1] 李贵刚[1] 李珂[1] 张虹[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院眼科,武汉430030

出　　处：《中华眼底病杂志》2007年第6期433-437,共5页Chinese Journal of Ocular Fundus Diseases

摘　　要：目的观察米诺环素对压力系统作用下大鼠视网膜神经细胞（RNs）的活性及凋亡的影响，探讨其对受损RNs保护的可能机制。方法制备大鼠RNs体外加压培养模型，通过细胞形态学观察、四唑盐（MTT）比色法测定细胞活力、吖啶橙／溴化乙锭（AO／EB）染色法检测细胞凋亡率等方法观察不同浓度的米诺环素对上述损伤细胞的保护作用，并用免疫细胞化学法观察iNOS和caspase-3表达的改变。结果加压培养后RNs与对照组相比形态改变较明显，细胞活力降低，53．93％的细胞发生凋亡。20．000μmol／L的米诺环素治疗组则细胞形态改善，活力显著增高，17．29％的细胞发生凋亡；免疫细胞化学法显示细胞内iNOS和caspase3表达较加压损伤组减少。结论一定剂量的米诺环素在体外可有效抑制压力引起的大鼠RNs损伤及凋亡；抑制iN0s和caspase-3的表达可能是其潜在作用机制。Objective To observe the effects of minocycline to the viability and apoptosis of rat＇s retinal neuron cells （RNC） under pressure, and to investigate the neuroprotective mechanisms of minocycline against the RNC damage. Methods Establish a model of rat＇s RNs under pressure cultured in vitro, the protective effect of minocycline is observed by different methods, including observing the morphology of the cells, evaluating the cells＇ viability by methyl thiazolyl tetrazolium （MTT） colorimetry assay, and detecting the cellular apoptosis with acridine orange/ethidium bromide （AO/EB） double staining by fluorescence microscopy. Immunocytochemistry was used to detect the expression of iNOS and caspase-3 in the cells. Results Obvious morphology changes of RNC were found in cells under pressure compared with the control; the viability of RNC decreased and cellular apoptosis was found in 53. 93% cells. The cellular morphology improved in the cells treated by 20μmol/L minocycline, the cellular viability significantly increased, and the cellular apoptosis was found in 17. 29% cells. In addition, the expression of iNOS and caspase 3 in the treated cells decreased compared with which in the pressured group. Conclusion Minocycline with a certain concentration can effectively inhibit pressure-induced damage and apoptosis of RNC of rats, and the inhibitory effect on expression of iNOS and capases-3 may be the underlying mechanism.

关 键 词：米诺环素 神经元 细胞凋亡 高眼压 动物实验 

分 类 号：R686[医药卫生—骨科学]