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目的 探讨银杏内酯 (Gin)对培养的皮层神经细胞的抗缺氧效应及其机制。方法 对原代培养的小鼠皮层神经细胞 ,进行随机分组 ,每个实验均分为正常对照组、缺氧组和 Gin组 ,每组做 8份重复测试。通过电镜观察神经细胞形态结构变化、利用甲基四唑蓝 (MTT)比色法检测细胞活性、速率法测定乳酸脱氢酶 (L DH)释放量、黄嘌呤氧化酶法检测超氧化物歧化酶 (SOD)活性、硫代巴比妥酸法测定丙二醛 (MDA )含量、硝酸还原酶比色法测定一氧化氮 (NO)含量 ,观察 Gin对缺氧神经细胞的影响。结果 (1)电镜观察显示 ,缺氧组神经细胞呈变性至坏死等不同程度的损伤 ;Gin组神经细胞细胞膜完整 ,微绒毛清晰可见 ,线粒体结构保存较好 ,与未缺氧的对照组相似。 (2 )缺氧后神经细胞活性 (0 .6 0 6±0 .0 35 )较正常对照组 (0 .6 77± 0 .0 2 8)降低 ,L DH释放量 (5 9.333± 9.5 2 1) U· ml- 1 · min- 1 较正常对照组 (38.5 0 0± 11.82 8) U· ml- 1 · min- 1 升高 ,其差异均有显著性意义 ;而预先给予 Gin的神经细胞活性(0 .6 4 5± 0 .0 2 7)明显高于缺氧组 ,L DH释放量 (40 .0 0 0± 6 .812 ) U· m l- 1 · m in- 1 显著低于缺氧组。 (3)缺氧后神经细胞 SOD活性 (46 .4 73± 7.4 72 ) NU / mg蛋白明显低于正常对照组 (5 5
Objective To investigate the anti-hypoxia effect of ginkgolide (Gin) on cultured cortical neurons and its mechanism. Methods The primary cultured mouse cortical nerve cells were randomly divided into three groups. Each experiment was divided into normal control group, hypoxia group, and Gin group. Each group was tested in 8 replicates. The morphological changes of nerve cells were observed by electron microscopy, the activity of cells was measured by methyltetrazolium blue (MTT) colorimetric method, the release rate of lactate dehydrogenase (L DH) was measured by rate method, and the superoxide dismutase was measured by xanthine oxidase method. (SOD) activity, thiobarbituric acid method to determine malondialdehyde (MDA) content, nitrate reductase colorimetric determination of nitric oxide (NO) content, observe the effect of Gin on hypoxic nerve cells. Results (1) Electron microscopy showed that nerve cells in the hypoxic group exhibited varying degrees of damage, such as degeneration to necrosis; in the Gin group, the neuronal cell membrane was intact, the microvilli were clearly visible, and the mitochondrial structure was well preserved, similar to the control group without hypoxia. (2) The activity of neurons after hypoxia (0.66±0. 0 35) was lower than that of the normal control group (0.67±0.028), and the release of LDH (5 9.333± 9.5 2 1) Compared with the normal control group (38.5 0 ± 11.82 8) U·ml-1 · min-1, U·ml-1 · min-1 was significantly higher than the normal control group; however, the neuronal activity of pre-administrated Gin was significantly increased. (0.64 5 ± 0. 0 2 7) was significantly higher than the hypoxic group. L DH release (40. 0 0 0 ± 6 .812) U · m l - 1 · m in - 1 was significantly lower than that of the hypoxic group. Oxygen group. (3) Neuronal SOD activity after hypoxia (46.47 73± 7.4 72) NU / mg protein was significantly lower than that of normal control group (5 5