目的：研究可乐定对原代培养大鼠皮质神经元氧糖剥夺( OGD)损伤的保护作用。方法取培养8 d的皮质神经元,分为正常对照组、模型对照组、可乐定(1.0,3.0,10.0μmol·L-1)预处理组。神经元氧糖剥脱损伤模型通过化学性缺氧、孵育液缺糖的方法建立。神经元损伤程度采用噻唑蓝( MTT)染色法和检测乳酸脱氢酶( LDH)的释放量来进行评价,观察预给予可乐定(1.0,3.0,10.0μmol·L-1)对神经元损伤的保护作用。结果显微镜下,正常对照组细胞密集,胞体饱满,边缘光滑,有较强折光性；神经元存活率(100.00±32.12)％,LDH释放比率(100.00±37.51)％。模型对照组细胞核固缩,细胞膜不完整,折光性差,MTT染色吸光度值明显降低,神经元存活率(53.61±7.62)％,LDH释放量显著增加,释放率为(166.07±9.65)％。可乐定(1.0,3.0,10μmol·L-1)预处理可明显逆转ODG损伤所致细胞形态的改变,剂量依耐性升高MTT染色吸光度值,神经元存活率分别为(67.53±10.54)％,(71.50±9.79)％和(87.48±5.29)％,同时可明显降低LDH的释放量,释放率分别为(136.45±25.72)％,(130.92±24.94)％和(121.63±32.68)％。结论可乐定对原代培养大鼠皮质神经元ODG损伤具有良好的保护作用。%Objective To determine the neuroprotective effect of clonidine on primary cultured cortical neurons in rats exposed to oxygen-glucose deprivation ( OGD) injury. Methods Cortical neurons cultured for 8 days were randomly assigned to the three groups: normal control group, model control group, and clonidine pretreatment group. OGD injury model was established by chemical hypoxia and glucose deprivation in incubation liquid for 4 h. Clonidine (1. 0, 3. 0, 10 μmol·L-1 ) was added 24 h before OGD injury. Neuronal injury was evaluated by MTT staining and the release of lactate dehydrogenase ( LDH) . Results Under the microscope, primary cultured cortical neurons in normal control group presented great density, round size, smooth edge, and high diopter,The suvival rate of neurons and the percentage of LDH releasing were (100. 00±32. 12)% and (100. 00 ± 37. 51 )%, respectively. After exposure to OGD injury, cortical neurons showed karyopyknosis, incomplete cell membranes, low diopters and a significant reduction in optical density of MTT staining. In addition, the suvival rate of neurons and the percentage of LDH releasing were (53. 61±7. 62)% and (166. 07±9. 65)% separately compared with normal control group. In the group with pretreatment of different concentrations of clonidine (1. 0, 3. 0, 10μmol·L-1), morphological changes induced by OGD injury were significantly reversed and optical density of MTT staining was dose-dependently raised. The percentages of survival neurons much higher than that of model control group were [(67. 53±10. 54)%, (71. 50±9. 79)% and (87. 48±5. 29)%, separately] and the obvious reductions of LDH releasing were [(136. 45±25. 72)%, (130. 92±24. 94)%and (121. 63±32. 68)%, respectively]. Conclusion Clonidine can exert neuroprotection against OGD-induced injury in primary cultured cortical neurons in rats.
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