绞股蓝皂苷对动脉粥样硬化的分子靶点及作用机制

Molecular targetsof action of gypenosides in atherosclerosis and its mechanism

目的探讨绞股蓝皂苷(GPs)对动脉粥样硬化(AS)的分子靶点及作用机制。方法通过中药系统药理学分析平台(TCMSP)数据库获取绞股蓝有效成分靶点,从基因表达综合数据库(GEO)中获取AS基因表达芯片并进行差异分析,对“药物靶点-疾病差异基因”交集基因PPI蛋白互作网络及Gene Ontology(GO)富集分析,获得GPs作用AS的关键靶点,并进行蛋白免疫印迹法(Western blot)实验验证;最后采用分子对接技术,将GPs单体分别与B淋巴细胞瘤-2蛋白(Bcl-2)、BCL2-Associated X蛋白(Bax)及半胱氨酸蛋白酶-3(Caspase-3)进行对接。结果网络药理学联合芯片分析筛选获得“绞股蓝-AS差异基因”交集基因中的凋亡靶点Bcl-2,通过Western blot进行实验验证;与模型组相比绞股蓝组Bax表达下调、Bcl-2表达上调、Bcl-2/Bax表达比值升高及Caspase-3表达下调,差异有统计学意义(P<0.05或<0.01);分子对接结果中,绞股蓝皂苷XXXV(Gypenoside XXXV)与Bcl-2结合能为-9.8 kcal/mol。结论GPs可能通过抑制细胞凋亡,从而预防AS的发生发展。

Objective To explore molecular targets and mechanism of action of Gypenosides(GPs)in atherosclerosis(AS).Methods Active ingredient targets of GPs was obtained from traditional Chinese medicine system pharmacology analysis platform(TCMSP)database.Differential gene expression(DEG)analysis was performed on an AS gene expression chip obtained from Gene Expression Omnibus database(GEO)to get AS-associated DEGs.Protein-protein interaction(PPI)network and Gene Ontology(GO)enrichment analyses were run on intersected genes between active ingredient targets of GPs and AS-associated DEGs to obtain key targets of GPs action on AS,which were verified by Western blot.Finally,Autodock was applied to examine whether GP monomer was able to dock with B lymphoma-2 protein(Bcl-2),BCL2-Associated X protein(Bax),cysteine protease-3(Caspase-3).Results Bcl-2,a key target of GPs,was verified by Western blot.When compared to model group,GP group had downregulated Bax but upregulated Bcl-2,increased Bcl/Bax ratio,and decreased Caspase-3 expression(P<0.05 or P<0.01).Autolock analysis showed that the binding energy of Gypenoside XXXV to Bcl-2 was-9.8 kcal/mol.Conclusion GPs may prevent AS occurrence and development by inhibiting apoptosis.