The Study Of Mechanism Based On Systematic Pharmacology Of Sanhuang Combination (Rhizoma Coptidis、Scutellariae Radix、Cortex Phellodendri) For Treating Non-Small Cell Lung Cancer

Object:With the in-depth of the research on the TME,the complex components of the TME and the interactions between the components have been understood.Targeting the tumor microenvironment has become an emerging therapy for tumor treatment.Some studies have shown that Traditional Chinese Medicine can improve immunity and anti-tumor effects.Therefore,this study explored the immune-related mechanism of Sanhuang combination(Rhizoma Coptidis,Scutellariae Radix,Cortex Phellodendri)for non-small cell lung cancer by system pharmacology,and comprehensive evaluation of the anti-tumor efficacy of the components in vivo and in vitro.Methods:Firstly,the database of the active ingredients of Sanhuang was constructed by using TCMSP,and the active compounds were screened by OB,DL and HL pharmacokinetic parameters.Then,WES and Sys DT methods were used to predict potential targets of active ingredients,and GOBP/KEGG pathway enrichments of the obtained targets were enriched,and compound-target network diagram were than built following that.Next,berberine,baicalin and phelloderine(BBP),the main active components of Sanhuang,were selected for in vivo and in vitro antitumor study.Firstly,CCK-8 and WB were used to verify the antitumor effect of BBP in vitro,then the in vivo antitumor effect of BBP was determined by pharmacodynamics experiment in tumor-bearing mice,and the content of CD8~+T cells infiltrated in tumor tissue was detected by flow cytometry.Finally,the anti-tumor efficacy of BBP combined with anti-PD-L1antibody was studied in tumor-bearing mouse model to explore the role of BBP in immunotherapy.Results:At first,56 active ingredients and 338 corresponding targets were obtained through systematic pharmacology analysis of Sanhuang.GOBP and KEGG enrichment analysis showed that Sanhuang was associated with non-small cell lung cancer and anti-PD-L1,suggesting that Sanhuang might play an anti-tumor effect by participating in tumor immune response.In vitro experiments showed that BBP could inhibit the proliferation of lung cancer cell lines and reduce the expression of cancer-promoting factors.In vivo studies have shown that BBP can inhibit tumor volume and prolong the survival of tumor-bearing mice by increasing the killing effect of CD8~+T cells in TME and restoring anti-tumor immune response.Moreover,BBP combined with anti-PD-L1 antibody increased the infiltration of CD8~+T cells and M1 macrophages in TME,and enhanced the anti-tumor effect of anti-PD-L1.Conclusion:BBP can inhibit the proliferation of NSCLC by increasing the killing effect of killer CD8~+T cells on tumor cells in TME.It has sensitized anti-tumor effect with anti-PD-L1,laying a foundation for the combination of Traditional Chinese medicine and anti-PD-L1.