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目的评价脑蛋白水解物对H_2O_2诱导的PC12细胞损伤的修复效果,建立一种脑蛋白水解物注射液的生物活性检测方法。方法以脑蛋白水解物的进口代表药物注射用脑蛋白水解物为例,采用PC12细胞培养,分析不同细胞接种浓度、不同浓度的H_2O_2损伤细胞、不同浓度脑蛋白水解物对抗干预的影响;采用MTT比色法对样品组、对照组和损伤组的细胞进行染色后,用酶标仪测定OD值,计算修复率,以评价脑蛋白水解物对模型细胞PC12损伤的保护作用;同时将该损伤修复评价方法用于国产脑蛋白水解物的生物活性检测。结果在8×10~4~12×10~4·mL~(-1)细胞接种量、0.5 mmol·L~(-1)H_2O_2损伤浓度下、60μg·L-1药物浓度(以含氮量计)下可成功的建立H_2O_2诱导的PC12细胞损伤模型。结论采用该方法对8批国产脑蛋白水解物注射液样品及注射用脑蛋白水解物进行对比检测,发现对H_2O_2损伤的细胞均具有良好修复作用。
Objective To evaluate the effect of brain protein hydrolyzate on the repair of H 2 O 2 -induced injury of PC12 cells and to establish a method for the bioassay of brain protein hydrolyzate injection. Methods The brain protein hydrolyzate was taken as an example to represent the injection of brain protein hydrolysates. The cultured PC12 cells were used to analyze the effects of different concentrations of cell seeding, different concentrations of H 2 O 2 injured cells and different concentrations of proteoglycan on the intervention. MTT After staining the cells in the sample group, the control group and the injury group by colorimetric method, the OD value was measured with a microplate reader and the repair rate was calculated to evaluate the protective effect of the brain protein hydrolyzate on the PC12 injury in the model cells; at the same time, Evaluation method for the biological activity of domestic brain protein hydrolyzate detection. Results The results showed that the drug concentration of 60 μg · L-1 at the dose of 8 × 10 ~ 4 ~ 12 × 10 ~ 4 · mL ~ (-1) and the concentration of 0.5 mmol·L ~ (-1) H_2O_2 Meter) successfully established H 2 O 2 -induced PC12 cell injury model. Conclusion The method of 8 batches of domestic brain protein hydrolyzate injection samples and injection of brain protein hydrolyzate were compared and found H_2O_2 injury cells have a good repair.