Network pharmacological combined with in vitro experimental to explore the molecular mechanism of Jinshui Liujun Decoction in adjuvant radiotherapy for malignant melanoma
Objective To explore the molecular mechanism of Jinshui Liujun Decoction in the treatment of malignant melanoma(MM)with adjuvant radiotherapy in combination with in vitro experiments.Methods The active ingredients and targets of Jinshui Liujun Decoction were obtained from TCMSP database,and the MM-related targets were obtained from GeneCards,TTD and OMIM databases with"malignant melanoma"as the keywords,so as to obtain the intersection targets of active ingredients and MM of Jinshui Liujun Decoction.The protein-protein interactions(PPI),network was obtained through STRING database,the gene ontology analysis(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)were conducted using Metascape database,and molecular docking was performed in AutoDockTools software;MTT method was used to detect the effect of Jinshui Liujun Decoction-containing serum on the proliferative activity of mouse B16 melanoma cells;Western blotting was used to detect the effect of Jinshui Liujun Decoction-containing serum on the regulation of protein expression of P38,p-P38,JNK,p-JNK,Beclin1,P62,and LC3;and the effect of Jinshui Liujun Decoction-containing serum on autophagy of B16 cells was observed by transmission electron microscopy.Results A total of 248 targets related to Jinshui Liujun Decoction,2 147 targets related to MM,and 149 intersecting targets were obtained;GO showed that the main biological processes were DNA binding to transcription factors,kinase binding,cytokine receptor binding,etc.;KEGG showed that it was mainly the cancer pathway and the MAPK signaling pathway;and the topology analysis of the network showed that quercetin was the main active ingredient in the treatment of MM by Jinshui Liujun Decoction,The network topology analysis showed that the main active ingredients of Jinshui Liujun Decoction for MM were quercetin,kaempferol,naringenin,etc.,and the main core targets were AKT1,TP53,JUN,etc.Docking between the core targets and the active ingredients showed that the binding activity was better.The MTT results showed that,compared with the blank group,the serum of the Jinshui Liujun Decoction containing the drug could inhibit the proliferation of B16 cells in a concentration-dependent manner.Western blotting showed that compared with the blank group,the medium and high dose groups could down-regulate the expressions of p-P38,p-JNK and P62 proteins,and up-regulate the expressions of LC3Ⅱ,LC3Ⅱ/LC3 Ⅰ and Beclin1 proteins in B16 cells.Transmission electron microscopy showed that the serum containing Jinshui Liujun Decoction promoted autophagy and reduced melanosome production in B16 cells.Conclusion Jinshui Liujun Decoction has many active ingredients and drug targets,and its targets and pathways for the treatment of MM are numerous.The molecular mechanism may be to inhibit the activation of the P38 pathway and JNK pathway by down-regulating the protein expression of p-P38,p-JNK,and P62,and up-regulating the protein expression of Beclin1,and LC3 Ⅱ/LC3 Ⅰ,thus inhibiting the proliferation of B16 cells;at the same time,it promotes apoptosis,activates cell autophagy,and plays a role in inhibiting the growth of melanosomes.
维普
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