| Objective: To study the material basis and mechanism of the anti-tumor effect of Sanjie Xiaoliu prescription on breast cancer by using network pharmacology.The key targets of Sanjie Xiaoliu prescription on breast cancer were screened out,and the correlation Network diagram was constructed using the analysis of Protein Protein Interaction Network(PPI).The material basis and mechanism of Sanjie Xiaomu prescription’s anti-tumor effect on breast cancer were revealed from the molecular level.The proliferation inhibition of MDA-MB-231 cells treated with Sanjie Xiaomufang was studied.The results showed that Sanjie Xiaoliu Fang had an effect on the polarization of macrophages by treating macrophages with conditioned medium.Methods: The key targets of the anti-tumor effect of Sanjie Xiaoliu prescription on breast cancer were obtained through network pharmacology technology,the network diagram of Sanjie Xiaoliu prescription and breast cancer target was constructed,and the material basis and mechanism of the anti-tumor effect of Sanjie Xiaoliu prescription on breast cancer were analyzed through PPI.To study the anti-tumor mechanism of Sanjie Xiaoliu prescription on breast cancer.MDA-MB-231 cells were selected,and the proliferation inhibition of MDA-MB-231 cells by Sanjie Xiaoliu Formula was detected by CCK8 method.Human monocyte THP cells were induced to polarize into M0 macrophages using phaboester(75ng/m L,72h),and the cell phenotype was examined by flow cytometry.MDA-MB-231 cells at logarithmic growth stage were cultured in Sanjie Xiaoliu prescription group and blank serum group for 48 h,and the supernatant was collected after 48 h to prepare conditional medium.Mda-mb-231-related macrophages were obtained by mixing the above prepared conditioned medium with RPMI-1640 in equal volume,incubating and inducing THP cells for 48 h.The expressions of CD86,CD206 and CD163 in MDA-MB-231 related macrophages were detected by Western blot as markers for M1-type tumor associated macrophages and M2-type tumor associated macrophages.Results: The pharmacological results of Sanjie Xiaoliu prescription on the anti-tumor effect of breast cancer showed that: There are 196 key targets for the anti-tumor effect of Sanjie Xiaoliu prescription on breast cancer,and the main active components include beta-sitosterol,Licorice Chalcone,Luteolin,Quercetin,etc.Core targets included P53(Tumor Protein p53,TP53),Estrogen Receptor 1(ESR1),Serine/threonine Kinase 1(AKT1),Oncogene C-myc,MYC,C-Jun N-terminal kinase(JUN),etc.Participate in hormone response,inorganic reaction,positive regulation of cell death and other biological processes,regulate cancer pathways,lipid and atherosclerosis,PI3K/Akt,hepatitis B and other pathways,play an anti-breast cancer therapeutic role.Experimental study on the anti-tumor mechanism of Sanjie Xiaoliu prescription on breast cancer results showed that the IC50 calculated by CCK8 method(26.510 for 24 h,20.419 for 48 h,15.884 for 72h)was the basis for determining the experimental grouping,which was as follows:Fetal bovine 10%,medicated serum 10%,medicated serum 20%,blank serum 10%,blank serum20%.The induced THP macrophages were detected by flow cytometry to determine the transformation of THP macrophages into M0 type macrophages and confirm the successful induction of phaboester.Western blot analysis showed that compared with the blank serum group,the cell surface antibody CD206 and CD163 were significantly down-regulated in the drug-containing serum group(P < 0.01),and the expression of CD68 was significantly higher(P< 0.01).The effect of 10% drug containing serum group was stronger than that of 20% drug containing serum group(P < 0.01).Conclusion: The mechanism of anti-tumor action of Sanjie Xiaoliu prescription on breast cancer was revealed through network pharmacology.It was found that Sanjie Xiaomufang inhibited the proliferation of MDA-MB-231 cells.Sanjie Xiaoliu prescription can inhibit the M2-type polarization of breast cancer-related macrophages,and can polarize M0 type macrophages to M1 type. |
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