摘要
研究天然化合物紫檀芪(pterostilbene,PTE)通过调控细胞焦亡及凋亡途径发挥抗脑胶质瘤的作用,通过网络药理学及分子对接方法分析PTE抗脑胶质瘤的可能通路及靶点。通过网络药理学筛选PTE作用靶点和脑胶质瘤靶点,构建靶点网络及蛋白-蛋白相互作用(PPI)网络,预测PTE可能作用靶点,通过AutoDock对核心靶点进行分子对接,通过富集分析预测PTE抗胶质瘤的作用途径;通过CCK-8法检测PTE对胶质瘤细胞U87MG、GL261细胞活力的影响;使用不同浓度PTE处理U87MG、GL261细胞,通过克隆形成实验检测PTE对细胞增殖的影响;划痕实验检测不同浓度PTE对胶质瘤细胞迁移能力的影响;采用Hoechst染色法观察PTE诱导胶质瘤细胞凋亡情况;采用JC-1染色法检测线粒体膜电位变化;通过倒置显微镜及乳酸脱氢酶(lactate dehydrogenase,LDH)释放实验观察PTE对胶质瘤细胞的焦亡诱导作用,Hoechst 33342/PI双染色法检测胶质瘤细胞膜的完整性;Western blot检测PTE诱导胶质瘤细胞后的细胞焦亡及凋亡相关蛋白表达变化。该研究共获得PTE抗脑胶质瘤的靶点37个,富集分析提示PTE通过多种信号途径包括癌症途径、癌症中蛋白聚糖、PI3K/AKT通路、凋亡调节通路等途径发挥抗脑胶质瘤的作用;分子对接显示PTE与主要靶点之间具有良好的结合活性;与空白组相比,PTE能显著降低U87MG、GL261细胞活性;与空白组相比,PTE组细胞的增殖、迁移、贴壁能力显著降低;与空白组比较,PTE可以诱导2种胶质瘤细胞凋亡,并且随药物浓度升高作用越明显;与空白组比较,PTE可以诱导U87MG细胞线粒体膜电位下降,并且随药物浓度升高作用越明显;与空白组相比,PTE组胶质瘤细胞出现焦亡特异性外观增多和LDH释放逐渐增加;Hoechst 33342/PI染色显示,与空白组相比,PI阳性细胞数量随着PTE浓度增加而显著增多;与空白组比较,PTE组凋亡相关蛋白活化聚(ADP-核糖)聚合酶1(cleave
This study aimed to explore the anti-glioma effect of natural compound pterostilbene(PTE) through regulating pyroptosis and apoptosis pathways,and to analyze the possible anti-glioma pathways and targets of PTE by network pharmacology and molecular docking.In this study,the action targets of PTE and the glioma targets were obtained by network pharmacology to construct a target network and a protein-protein interaction(PPI) network to predict the possible action targets of PTE against glioma.Molecular docking was performed on the core targets by AutoDock and the action pathways of PTE against glioma were predicted by enrichment analysis.In addition,the effect of PTE on the viability of U87MG and GL261 glioma cells was detected by CCK-8 assay.Clone formation assay and cell scratching assay were used to explore the effect of different concentrations of PTE on the proliferation and migration,respectively of glioma cells.Hoechst staining was used to observe PTE-induced apoptosis in glioma cells.The changes in mitochondrial membrane potential were detected by JC-1 staining.The pyroptosis-inducing effect of PTE on glioma cells was observed by inverted microscopy and lactate dehydrogenase(LDH) assay.Hoechst 33342/PI dual staining assay was performed to detect the integrity of glioma cell membranes.The expressions of pyroptosis and apoptosis-related proteins in glioma cells after PTE induction were determined by Western blot.In this study,37 anti-glioma targets of PTE were obtained,and enrichment analysis suggested that PTE exerted anti-glioma effects through various signaling pathways including cancer pathway,proteoglycan in cancer,PI3K/AKT pathway,and apoptosis regulatory pathway.Molecular docking revealed that PTE had good binding activity with the main targets.Compared with the control group,PTE significantly reduced the viability as well as the proliferation,migration and adhesion abilities of U87MG and GL261 cells;it induced the apoptosis of the two glioma cells and the decrease of mitochondrial membrane potential i
作者
陈卓
方兴刚
郭兴荣
栾思琪
华娇
罗杰
CHEN Zhuo;FANG Xing-gang;GUO Xing-rong;LUAN Si-qi;HUA Jiao;LUO Jie(Clinical College of Traditional Chinese Medicine,Hubei University of Chinese Medicine,Wuhan 430065,China;Hubei Key Laboratory of Embryonic Stem Cell Research,Shiyan 442099,China;Taihe Hospital,Affiliated Hospital of Hubei University of Medicine,Shiyan 442099,China;Taihe Hospital,Doctoral Training Unit of Hubei University of Chinese Medicine,Shiyan 442099,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2023年第13期3589-3601,共13页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(82073232,81671831)
湖北省引进国外人才和智力项目(Yz2020015)
湖北省卫生健康委青年人才项目(WJ2021F032)。
关键词
紫檀芪
细胞焦亡
细胞凋亡
脑胶质瘤
焦孔素E(GSDME)
网络药理学
pterostilbene
pyroptosis
apoptosis
glioma
gasdermin E(GSDME)
network pharmacology D0I:10.19540/j.cnki.cjcmm.20230216.703
