A Network Pharmacology-based Approach To Explore The Active Ingredients And Molecular Mechanism Of Aidi Injection Anti-prostate Cancer

Objective:To explore the active ingredients and molecular mechanism of AIDI Injection(hereinafter referred to as "AIDI")anti-prostate cancer(PCa),so as to provide a preliminary research basis for the clinical application of AIDI Injection in the treatment of PCa.Method : CCK-8 method and colony formation assay were used to detect the effect of AIDI on the proliferation and colony formation of PC3 and DU145 cells.Flow cytometry was used to detect the effect of AIDI on DU145 cell cycle and apoptosis.The effects of AIDI on the migration and invasion of PC3 and DU145 cells were detected by wound healing and Transwell method.Network pharmacology method was used to predict and analyze the active ingredients,targets and related signaling pathways of AIDI in inhibiting PCa.The predicted key targets and related signaling pathways were verified by Western blot.Molecular docking technology was used to predict the potential active ingredients of AIDI anti-prostate cancer.Finally,the chemical constituents of AIDI were analyzed by UPLC-Q-TOF/MS.Results:AIDI significantly inhibited the proliferation of PC3 and DU145 cells in a concentration and time-dependent manner.AIDI could inhibit the colony formation of PC3 and DU145 cells with a concentration dependent manner.AIDI could induce DU145 cells apoptosis and block DU145 cells in G2 / M phase.AIDI could significantly inhibit the migration and invasion of PC3 and DU145 cells.A total of 36 potential core targets of AIDI against PCa were obtained by network pharmacology analysis,corresponding to 128 chemical components in AIDI,which were enriched to 158 signaling pathways.The top10 signaling pathway includes MAPK,PI3 K Akt and HIF-1 etc.Western Blot results showed that AIDI could markedly up-regulate the expression of p-JNK,p-p38,p-ERK and ERK in DU145 cell.The results of molecular docking showed that JNK had good binding with kaempferol,and p38 had good binding with(z)-1-(2,4-dihydroxyphenyl)-3-(4-hydroxyphenyl)prop-2-en-1-one,7-o-methylsulfonatol,calycosin and n-salicylidene-salicylamine,respectively.A total of 31 chemical constituents were identified by UPLC-Q-TOF/MS.Conclusion : AIDI could inhibit the growth and metastasis of PCa cells via increasing the activity of MAPK signaling pathway and inducing apoptosis and cycle arrest of PCa cells.Kaempferol,(z)-1-(2,4-dihydroxyphenyl)-3-(4-hydroxyphenyl)prop-2-en-1-one,7-o-methylmalonulatol,calycosin and nsalicylidene-salicylamine might be the active components of AIDI anti-PCa.