Mechanisms Of Zuojin Pill Component Combined With EcN Inducing Apoptosis Of Colorectal Cancer Cells

Objective The active ingredients and thrie functional targets of Zuojin Pill on colorectal cancer were analyzed by network pharmacology.Fourther more,the effect of Zuojin pill combined with probiotic Ec N on colon cancer cells and their potential mechanisms were investagated.Methods(1)The active ingredients in Zuojin Pills and their functional targets of in anti-color ectal cancer were analyzed by network pharmacological.The active ingredient-functional target-parthway network of Zuojin pill was constructed via gene function annotation analysis.The potential target and their pathway of Zuojin Pill antianti-colorectal cancer were predicted by enrichment of analysis.The active ingredients of Zuojin pill were screened preliminarily and were used for further investigation.(2)Berberine,evodiamine,quercetin,isorhamnetin and isorhamnetin were co-cultured with CT26 cells mouse colorectal cancer cells,and inhabition of the those compounds to CT26 cells growth was evaluated by MTS method.Further more,the effect of berberine and evodiamine combined with probiotic Ec N on the groth of CT26 cells were evaluated by MTS.Enzyme-linked immunosorbent assay(ELISA)was used to detect the secretion of IL-17,IL-22 and IL-23.To observe the effects of berberine and evodiamine combined with probiotic Ec N induced apoptosis of CT26 cells,the death of cells was analyzed by flow cytometry through Annexin V-FITC/PI double-staining of CT26 cells.Hoechst33258 staining was used to analyze the apoptosis of CT26 cells,and intestine tissue from colon cancer mouse treated with Zuojin Pills or its active ingredens alone or combined with probiotic Ec N.Western blot was used to detect the apoptosis related protiens in CT26 cells or intestine tissue with the same treatment condition as avobe.Results.Berberine and Evodiamine combined with probiotics Nissle 1917 induce the apoptosis on CT26 cells,and intestine tissue from chemicals induced colon cancer mouse model,which may be related to caspase3 involved with increase of Cleavedcapase3 and Bax/Bcl-2 ratio.The results in current study may provide a great potential in application of Zuojin pill combined with probiotics to treat coclon cancer.