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The study of the protective effect of panoxadiol on Alzheimer's disease cells based on network pharmacology
梁喜才姚璎珈王玉莹李秀丽王雅萌蔺 莹时 悦杨静娴
辽宁中医药大学药学院,辽宁 大连 116600
LIANG Xi-caiYAO Ying-jiaWANG Yu-yingLI Xiu-liWANG Ya-mengLIN YingSHI YueYANG Jing-xian
School of Pharmacy,Liaoning University of Traditional Chinese Medicine,Dalian Liaoning 116600,China
阿尔茨海默病 网络药理学 分子对接 人参二醇 FYN Ca2+超载
Alzheimer's disease(AD) network pharmacological analysis molecular docking PD FYN Ca2+ overload
R284.1; R329.25; R319; R348.1; R745.702.2; R745.701; R977.6
目的 运用网络药理学预测及体外验证方法,研究人参二醇(PD)对阿尔茨海默病(AD)细胞的保护作用。方法 运用网络药理学技术,筛选出107个治疗AD的方剂,筛选出现频率最高的人参及其AD作用的靶点,利用分子对接技术,寻找与非受体酪氨酸激酶(FYN)对接评分最高的成分。体外建立APP-N2a细胞模型,采用MTT法检测细胞存活率; LDH方法检测细胞的损伤程度; 流式细胞技术检测细胞凋亡及细胞内Ca2+浓度变化情况; Western blot检测磷酸化FYN蛋白表达情况。结果 通过网络药理学方法筛选出人参中18个活性成分和29个AD相关靶点,分子对接结果显示,PD与FYN有较强的结合性。实验证明,PD可增加细胞的存活率,降低LDH的释放,且可明显减少细胞凋亡,减轻AD细胞内Ca2+超载,降低FYN-Y416蛋白的表达。结论 通过实验验证了网络药理学的预测结果,PD对AD的保护作用可能与抑制FYN磷酸化有关。
Aim To explore the therapeutic effects of main active compounds of panaxadiol(PD)in on Alzheimer's disease(AD)via network pharmacological analysis and Mmolecular docking. Methods A total of 107 prescriptions for AD treatment were screened by using network pharmacology, screening for the highest frequency of ginseng and its target for AD. Use molecular docking technology was used to find components with the highest score for non-receptor tyrosine kinase(FYN)docking. Then we successfully estimatedestablished AD cell model with overexpressinged APP proteins in vitro. Next,the cell viability was detected by MTT assay,the cell damage was detected by LDH assay,the apoptosis and intracellular Ca2+ concentration were detected by flow cytometry,and phosphorylated FYN protein expression was detected by Western blot detection of.phosphorylated FYN protein expression. Results Eighteen active components of Gginseng and 29 AD-related targets were screened by the method of network pharmacology. The results of molecular docking showed that PD had strong binding effects with FYN. The results showed that PD could increase the survival rate of cells,reduce the release of LDH,reduce apoptosis,and improve AD cells' intracellular Ca2+ overload and reduce the expression of FYN-Y416 protein. Conclusion The experimental results of network pharmacology were are verified and the protective effect of PD on AD may be related to inhibition of FYN signaling pathway.


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基金项目:国家自然科学基金面上项目( No 81173580)
更新日期/Last Update: 2018-08-26