基于均匀设计实验的栀子-三七主要有效成分抗Aβ_(1-42)诱导SH-SY5Y细胞损伤的配伍研究  

Compatibility study of the main active components of Gardenia-Panax notoginseng against Aβ_(1-42)-induced damage to SH-SY5Y cells based on uniform design experiments

作　　者：岳钰茗 杨冰 周福军 王泽凡[2,3,4] 周钰通 付庆 单淇 徐旭 YUE Yu-ming;YANG Bingg;ZHOU Fu-jun;WANG Ze-fan;HOU Yu-tong;FU Qing;SHAN Qi;XU Xu(Tianjin University of Traditional Chinese Medicine,Tianjin 300193,China;National Key Laboratory of Drug Evaluation and Systematic Transformation(State Key Laboratory of Original Drug Release Technology and Pharmacokinetics),Tianjin 300301,China;Tianjin Key Laboratory of Quality Marker of Traditional Medicine,Tianjin 300301,China;Tianjin Institute of Pharmaceutical Research,Tianjin 300301,China)

机构地区：[1]天津中医药大学,天津300193 [2]药物成药性评价与系统转化全国重点实验室(原释药技术与药动学国家重点实验室),天津300301 [3]天津市中药质量标志物重点实验室,天津300301 [4]天津药物研究院,天津300301

出　　处：《中国新药杂志》2023年第18期1882-1892,共11页Chinese Journal of New Drugs

基　　金：释药技术与药动学国家重点实验室(天津药物研究院)开放课题资助(010161003)。

摘　　要：目的:采用均匀设计实验方法,探索栀子和三七中5个主要有效成分的有效配伍比例,为临床治疗阿尔茨海默病提供初步实验依据。方法:建立β-淀粉样蛋白(Aβ_(1-42))诱导SH-SY5Y细胞损伤模型,以细胞存活率为指标,研究主要有效成分抗Aβ_(1-42)诱导SH-SY5Y神经细胞损伤的有效浓度,以损伤恢复率为指标选出5种药效较好的成分。在此基础上筛选出的5种药效良好的成分分别为自变量X_(1),X_(2),X_(3),X_(4),X_(5),损伤恢复率(%)为因变量Y,选用U_(21)(21^(5))均匀设计表和线性、二次多项式2种数学模型,以逐步回归方法筛选变量,确定5种有效成分的有效配伍比例。结果:栀子-三七主要有效成分除京尼平外均能提高Aβ_(1-42)诱导SH-SY5Y细胞损伤的细胞存活率,并呈浓度依赖性。筛选得到5种药效良好的成分为三七皂苷R1、人参皂苷Rb1、人参皂苷Rg1、栀子苷、京尼平龙胆双糖苷。均匀设计实验结果经回归分析得到Y=12.77501992-0.4938270427X_(2)+0.07063466558X_(1)X_(2)-0.022126968533X_(1)X_(5)+0.05300069195X_(3)X_(4)(r=0.8574,F=11.1059,P=0.002),均匀设计实验验证得到最佳配伍比例为三七皂苷R1∶人参皂苷Rb1∶人参皂苷Rg1∶栀子苷∶京尼平龙胆双糖苷=1∶1∶1∶1∶0。行为学试验显示与模型组小鼠比较,盐酸多奈哌齐组、栀子-三七最优配伍的测试次数和记忆错误次数均显著降低。ROS和Western blot结果显示与模型组比较,盐酸多奈哌齐组、栀子-三七最优配伍可以有效降低ROS的荧光强度和RAGE蛋白的表达。结论:运用均匀设计筛选获得的最优配伍比例可有效改善阿尔茨海默病,其作用机制可能为抑制RAGE-ROS通路的激活发挥抗阿尔茨海默病的作用,为治疗阿尔茨海默病提供了实验依据。Objective:To explore the effective compatibility ratio of the 5 main active ingredients in Gardenia and Panax notoginseng by uniform design test method thus to provide preliminary experimental evidence for clinical treatment of Alzheimer's disease.Methods:A model of SH-SY5Y cell injury induced byβ-amyloid(Aβ_(1-42))was established,and the cell survival rate was used as an indicator to study the effective concentrations of the main active ingredients against Aβ_(1-42)-induced SH-SY5Y neuronal injury,and according to the damage recovery rate,five ingredients with better efficacy were selected.On this basis,the five components with good efficacy were used as independent variables X_(1),X_(2),X_(3),X_(4) and X_(5),respectively,the injury recovery rate(%)as the dependent variable Y,U_(21)(21^(5))uniform design table and 2 mathematical models of linear and quadratic polynomials were selected,and the variables were screened by stepwise regression method to determine the effective compatibility ratio of the 5 active ingredients.Results:The main active components of Gardenia-Panax notoginseng,except genipin,could increase the cellsurvival rate of SH-SY5Y cells induced by Aβ_(1-42) in a concentration-dependent manner.5 ingredients with good efficacy were identified as notoginsenoside Rl,ginsenoside Rbl,ginsenoside Rgl,geniposide,and genipin gentiobiglycoside.The results of uniform design experiments were obtained by regression analysis and Y=12.775 019 92-0.493 827 042 7X_(2)+0.070 634 665 58X_(1)X_(2)-0.022 126 968 533X_(1)X_(5)+0.0530 006 919 5X_(3)X_(4)(r=0.857 4,F=11.105 9,P=0.002),and the uniform design test verified that the best combination ratio was notoginsenoside Rl:ginsenoside Rbl:ginsenoside Rgl:geniposide:genipin gentiobiglycoside=1:1:1:1:0.Behavioral experiments showed that compared with the model group,the number of tests and memory errors of donepezil hydrochloride group and gardenia-Panax notoginseng optimal matching were significantly reduced.The results of ROS and Western blot showed that compared wi

关 键 词：阿尔茨海默病 栀子苷 三七皂苷R1 人参皂苷RB1 人参皂苷Rg1 配伍 

分 类 号：R965[医药卫生—药理学]