基于网络药理学和细胞实验探讨和厚朴酚对 PC12细胞HIF-1α-VEGF通路的调节作用

Study on regulation of honokiol on HIF-1α-VEGF pathway in PC 12 cells based on network pharmacology and experimental verification

基于网络药理学和细胞实验探讨和厚朴酚对PC12细胞HIF-1α-VEGF通路的调节作用。通过TCMSP数据库和Swiss Target Prediction数据库筛选出127个药物靶点;采用DisGeNET数据库收集到287个疾病靶点;通过String数据库构建PPI网络;采用DAVID数据库进行GO和KEGG富集分析。GO富集分析得到173个条目。KEGG通路筛选出前20条信号通路,包括HIF-1通路、VEGF通路、PI3K-Akt通路等。分子对接结果表明和厚朴酚与HIF-1α降解酶PHDs、VHL有较好的结合力。CCK-8法发现和厚朴酚能有效增加PC12细胞的存活率。正常环境下,和厚朴酚给药后HIF-1α、VEGF、PSD 95、SYN 1蛋白表达升高(P<0.05,P<0.01)。缺氧环境下,和厚朴酚给药后HIF-1α和VEGF蛋白表达显著降低(P<0.05,P<0.01),而PSD 95和SYN 1蛋白表达显著升高(P<0.01,P<0.001)。综上,正常和缺氧环境下和厚朴酚对PC12细胞HIF-1α-VEGF通路有相反的调节作用,为和厚朴酚抗脑缺血、抗抑郁和抗肿瘤机制研究提供可靠的理论和实验支持。

To study the regulation of honokiol on the HIF-1α-VEGF pathway in PC12 cells based on network pharmacology and experimental verification.To search 127 drug targets through the TCMSP database and the Swiss Target Prediction database,then using the DisGeNET database to collect 287 disease targets.Constructed PPI network map by String online database.At the same time GO and KEGG enrichment analyses were carried out by using the DAVID database.Go enrichment analysis obtained 173 entries.The KEGG pathway screened the top 20 signaling pathways,including the HIF-1 pathway,VEGF pathway,PI3K-Akt pathway,and others.The molecular docking results showed that honokiol bound well to HIF-1αdegrading enzymes PHDs,VHL.The CCK-8 method found that honokiol could effectively increase the survival rate of PC12 cells.Under normal conditions,HIF-1α,VEGF,PSD 95,and SYN 1 protein expression were elevated after honokiol administration(P<0.05,P<0.01).Under hypoxic conditions,HIF-1αand VEGF protein expression were significantly lower(P<0.05,P<0.01),and PSD 95 and SYN 1 protein expression were significantly higher after honokiol administration(P<0.01,P<0.001).In conclusion,honokiol has opposite regulatory effects on the HIF-1α-VEGF pathway,providing reliable theoretical and experimental support for the study of the anti-ischemia,antidepressant,and antitumor mechanisms of honokiol.