脑蛋白水解物对H2O2诱导的PC12细胞氧化损伤的修复作用及其胃漂浮片处方的优化

摘要

Objective:To detect the repair effect of brain protein hydrolyzate(BPH)on the oxidative damage induced by H2O2in the PC12 cells,and to optimize the prescription of BPH stomach floating tablets using the star design effect surface method.Methods:The PC12 cells in the logarithmic phase were divided into normal control group,model group(300 μmol·L-1H2O2),BPH group,artificial gastric juice-treated BPH(GBPH)group, artificial intestinal juice-treated BPH(IBPH)group,artificial gastric juice-treated BPH tablets(GBPH-T)group and artificial gastric juice-treated BPH floating tablets(GBPH-FT)group(The latter five groups were added with 20,40,60,80 and 100 mg·L-1BPH treated with different conditions);at the same time blank control group was set up.The PC12 cells in normal control group didn't receive any treatment,the blank control group was added with medium only,and the PC12 cells in model group were treated with H2O2(300 μmol·L-1)for 3 h. The cell viability was detected by MTT assay.Using Design-expert 8.0.6 Trial software,the dosages of HPMC-K4M,octadecanol and acrylic resin Ⅱ were used as the investigation factors,and the 8 h cumulative release in vitro was used as the evaluation index to optimize the prescription.Results:Compared with normal control group, the activity of PCl2cells in 60 mg·L-1BPH group was increased(P0.05).Compared with GBPH-T group, the cell viability in GBPH-FT group was significantly increased(P 8 h;the difference of the measured value of 8 h cumulative release and the predicted value was not statistically significant (P>0.05).Conclusion:BPH has a significant repair effect on the H2O2-induced oxidative damage in the PC12 cells.Star design-effect surface method has good predictability and reproducibility;it is reasonable and feasible, and can be used to optimize the prescription of BPH stomach floating tablets.%目的:检测脑蛋白水解物(BPH)对 H2O2诱导 PC12细胞氧化损伤的修复作用,运用星点设计-效应面法对 BPH 胃漂浮片的处方进行优化.方法:将对数生长期 PC12细胞分为正常对照组、模型组(300 μmol· L-1H2O2)、BPH组、人工胃液处理的BPH(GBPH)组、人工肠液处理的BPH(IBPH)组、人工胃液处理的BPH片(GBPH-T)组和人工胃液处理的BPH漂浮片(GBPH-FT)组(后5组均加不同条件处理的20、40、60、80和100 mg·L-1BPH),另设空白对照组;正常对照组不做任何处理,空白对照组只加培养基,其他各组采用300 μmol·L-1H2O2孵育3 h 造成损伤模型.采用 MTT 法检测细胞活力;利用 Design-expert 8.0.6 Trial软件,将 HPMC-K4M、十八醇和丙烯酸树脂Ⅱ号用量作为考察因素,以8 h累积释放度为评价指标,优化处方.结果:与正常对照组比较,60 mg·L-1BPH 组细胞活力明显升高(P0.05);与 GBPH-T 组比较,GBPH-FT组细胞活力明显升高(P8 h, 8 h累积释放度实测值与预测值比较差异无统计学意义(P>0.05).结论:BPH 对 H2O2诱导的 PC12细胞氧化损伤具有修复作用.星点设计-效应面法预测性和重复性良好,合理可行,可用于BPH胃漂浮片处方的优化.