脑苷肌肽对过氧化氢诱导的PC12细胞氧化应激损伤的抑制作用

Inhibition effect of encephalon glycoside on H_2O_2-induced oxidattive stress injury in PC12 cell

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 explore the inhibition effect of encephalon glycoside on H_2O_2-induced oxidattive stress injury in PC12 cell.Methods The cases were randomly divided into 5 groups,the normal group,model group（ 600 μmol·L^（-1）H_2O_2） and in the low,medium and high dose test groups（ 30,100,300 μg·m L^（-1） encephalon glycoside）.The viability of PC12 cell was measured by MTT assay. Cell apoptosis was detected by Hoechst staining method. The contents of malondialdehyde（ MDA）, superoxide dismutase（ SOD）, actate dehydrogenase（ LDH） and L- glutathione（ GSHx） and the activity of cysteinyl aspartate specific proteinase 3（ Caspase 3）,Caspase 9 were determinated by spectrophotometry. The expressions of B- cell lymphoma- 2（ Bcl-2）,Bcl-2 associated X protein（ Bax） were assayed by Western blot. The expressions of Bax,Bcl- 2,Caspase 3 and Caspase 9 were detected by reverse transcription polymerase chain reaction（ RT- PCR）.Results Compared with model group,cell viability [（ 62. 11 ± 5. 94） %vs（ 71. 48 ± 7. 16） %,（ 82. 44 ± 7. 39） %,（ 90. 66 ± 9. 10） ] increased（ P〈 0. 05） in three dose test groups. Compared with model group,cellapoptotic rate decreased（ P〈 0. 05）,the levels of MDA and LDH reduced（ P 〈0. 01）,the levels of SOD and GSHx increased（ P〈 0. 05）,the activity of Caspase 3 and Caspase 9 reduced（ P〈 0. 5）,the expression of Bax protein decreased（ P〈 0. 05）,the expressions of Bcl- 2 protein and mRNA increased（ P〈 0. 05）,the expressions of Bax,Caspase 3 and Caspase 9 mRNA decreased in three dose test groups. Conclusion The encephalon glycoside was resistance to H_2O_2-induced oxidattive stress in PC12 cell.