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目的研究脑苷肌肽对PC12细胞氧化应激损伤的抑制作用。方法 PC12细胞随机分为5组:正常组、模型组及3个剂量实验组(30,100,300μg·mL~(-1)脑苷肌肽)。模型组及实验组用600μmol·L~(-1)过氧化氢(H_2O_2)诱导PC12细胞氧化应激模型。噻唑蓝法及Hoechst染色分别检测细胞活力及凋亡;分光光度法检测丙二醛(MDA)、超氧化物歧化酶(SOD)、乳酸脱氢酶(LDH)、谷胱甘肽(GSHx)含量及天冬氨酸蛋白水解酶(Caspase 3)与Caspase 9活性;免疫印迹技术分析B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)的表达情况;反转录聚合酶连锁反应法检测各组细胞中Bcl-2、Bax、Caspase 3及Capase 9 mRNA表达。结果与模型组的细胞活力为(62.11±5.94)%比较,3个剂量实验组的细胞活力[(71.48±7.16)%,(82.44±7.39)%,(90.66±9.10)%]明显提高(P<0.05)。与模型组比较,3个剂量实验组细胞凋亡率明显降低(P<0.05),MDA及LDH含量明显降低(P<0.05),SOD及GSHx含量明显提高(P<0.05),Casoase 3及Caspase 9活性明显减弱(P<0.05),Bax表达量明显下调,Bcl-2表达量明显上调(P<0.05)。结论脑苷肌肽可抑制H_2O_2诱导的PC12细胞氧化应激损伤。
Objective To study the inhibitory effect of cerebroside on oxidative stress injury in PC12 cells. Methods PC12 cells were randomly divided into 5 groups: normal group, model group and 3 doses of experimental group (30, 100, 300μg · mL -1 cerebroside). Model group and experimental group induced oxidative stress in PC12 cells with 600μmol·L -1 hydrogen peroxide (H 2 O 2). Cell viability and apoptosis were detected by thiazolyl blue staining and Hoechst staining respectively. The contents of malondialdehyde (MDA), superoxide dismutase (SOD), lactate dehydrogenase (LDH) and glutathione (GSHx) were measured by spectrophotometry Caspase 3 and Caspase 9 were detected by Western blotting. The expression of Bcl-2 and Bcl-2 protein was analyzed by Western blotting. Enzyme linked reaction method was used to detect the expression of Bcl-2, Bax, Caspase 3 and Capase 9 mRNA in each group. Results Compared with the model group (62.11 ± 5.94%), the cell viability of the three experimental groups (71.48 ± 7.16%, (82.44 ± 7.39)%, (90.66 ± 9.10)%, P <0.05). Compared with the model group, the apoptotic rates of the three groups were significantly decreased (P <0.05), MDA and LDH contents were significantly decreased (P <0.05), SOD and GSHx contents were significantly increased (P <0.05), Casoase 3 and Caspase 9 activity was significantly decreased (P <0.05), Bax expression was significantly decreased, Bcl-2 expression was significantly increased (P <0.05). Conclusion Cerebroside can inhibit H 2 O 2 -induced oxidative stress injury in PC12 cells.