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

doi:10.13481/j.1671-587x.20180215

吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (02): 286-291.doi: 10.13481/j.1671-587x.20180215

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

杨昕祺, 王广红, 糜心雅, 安丽萍, 杜培革, 王曼力

北华大学长白山药用动植物活性多肽国家地方联合工程实验室, 吉林吉林 132013

收稿日期:2017-11-15 出版日期:2018-03-28 发布日期:2018-03-30
通讯作者: 杜培革,教授,硕士研究生导师(Tel:0432-64608278,E-mail:dupeige2001@126.com);王曼力,副教授(Tel:0432-64608278,E-mail:bhwangml@163.com) E-mail:dupeige2001@126.com;bhwangml@163.com
作者简介:杨昕祺(1993-),女,吉林省辽源市人,在读医学硕士,主要从事药物分析方面的研究。
基金资助:
吉林省科技厅科技计划项目资助课题（20160441001SC）

Repair effect of brain protein hydrolysate on H₂O₂-induced oxidative damage in PC12 cells and optimization of formulation of its stomach floating tablets

YANG Xinqi, WANG Guanghong, MI Xinya, AN Liping, DU Peige, WANG Manli

Laboratory of Changbai Mountain Medicinal Plants and Animals Active Peptide National Joint Engineering, Beihua University, Jilin 132013, China

Received:2017-11-15 Online:2018-03-28 Published:2018-03-30

摘要/Abstract

摘要： 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^-1 H₂O₂）、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^-1 H₂O₂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（P<0.05）；与模型组比较，BPH、IBPH、GBPH和GBPH-FT组细胞活力明显升高（P<0.05或P<0.01）；与BPH组比较，IBPH组细胞活力明显降低（P<0.05），GBPH组细胞活力差异无统计学意义（P>0.05）；与GBPH-T组比较，GBPH-FT组细胞活力明显升高（P<0.05）。优化处方为BPH 20 mg，乳糖10 mg，硬脂酸镁0.4mg，微晶纤维素20 mg，HPMC-K4M 27 mg，十八醇63 mg，丙烯酸树脂Ⅱ号13 mg；BPH胃漂浮片均在5 s内起漂，持续漂浮>8 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。

关键词: 漂浮片, 脑蛋白水解物, 细胞氧化损伤, 星点设计

Abstract: Objective:To detect the repair effect of brain protein hydrolyzate (BPH) on the oxidative damage induced by H₂O₂ in 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^-1 H₂O₂),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^-1 BPH 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 PCl₂ cells in 60 mg·L^-1 BPH group was increased (P<0.05). Compared with model group,the vitalities of PC12 cells in BPH group,IBPH group,GBPH group and GBPH-FT group were increased significantly (P<0.05 or P<0.01). Compared with BPH group,the cell viability in IBPH group was decreased significantly (P<0.05),and there was no significant difference in GBPH group (P>0.05). Compared with GBPH-T group,the cell viability in GBPH-FT group was significantly increased (P<0.05).The optimal prescription was BPH 20 mg,lactose 10 mg,magnesium stearate 0.4 mg,microcrystalline cellulose 20 mg,HPMC-K4M 27 mg,octadecanol 63 mg,acrylic resinⅡ13 mg; all floating tablets drift in 5 s and sustained floating > 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.

Key words: star design, brain protein hydrolyzate, floating tablets, cell oxidative damage

中图分类号:

R965.1

杨昕祺, 王广红, 糜心雅, 安丽萍, 杜培革, 王曼力. 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[J]. 吉林大学学报(医学版), 2018, 44(02): 286-291.

YANG Xinqi, WANG Guanghong, MI Xinya, AN Liping, DU Peige, WANG Manli. Repair effect of brain protein hydrolysate on H₂O₂-induced oxidative damage in PC12 cells and optimization of formulation of its stomach floating tablets[J]. Journal of Jilin University Medicine Edition, 2018, 44(02): 286-291.

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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

Repair effect of brain protein hydrolysate on H₂O₂-induced oxidative damage in PC12 cells and optimization of formulation of its stomach floating tablets

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