| Objective:To study the mechanism of ursolic acid(UA)in the treatment of hepatocellular carcinoma.Methods:(1)The main effective components of AiTongXiao were screened by network pharmacology methods,and the main pathways that AiTongXiao may act on the core target genes and enrichment of hepatocellular carcinoma were analyzed.Finally,the possible pathways that AiTongXiao may play the role of inhibiting hepatocellular carcinoma were obtained.(2)Bel-7402 cells were cultured and divided into control group(without drug)and experimental group(15μM-30μM ursolic acid)MTT assay was used to observe the effect of ursolic acid on the proliferation of Bel-7402 cells;The effect of ursolic acid on the colony formation of Bel-7402 cells was observed by colony formation assay;Hoechst 33342 staining was used to observe the effect of ursolic acid on apoptosis of Bel-7402 cells;The effect of ursolic acid on the apoptosis rate of Bel-7402 cells was observed by flow cytometry;RT-q PCR and WB were used to observe the effects of ursolic acid on the expression of P53、Bcl-2 and CASP3 mRNA and protein in Bel-7402 cells.Results:(1)Ursolic acid was selected as the main active component of AiTongXiao through network pharmacology,and further analysis showed that ursolic acid may act on the core target of hepatocellular carcinoma,because P53 signal pathway is also the main signal pathway enriched,It is speculated that ursolic acid may play a role in the treatment of hepatocellular carcinoma through P53/Bcl-2/CASP3 signaling pathway.(2)15μM-30μM UA had a significant inhibitory effect on hepatocellular carcinoma Bel-7402 cells,and was concentration-dependent;Compared with the control group,different concentrations of UA in the experimental group could inhibit the colony formation of Bel-7402 cells in varying degrees,and the colony formation gradually decreased with the increase of ursolic acid concentration.Compared with the control group,15μM-30μM ursolic acid can significantly promote cell apoptosis,and with the increase of UA concentration,apoptosis is more significant.The results of RT-q PCR and WB showed that ursolic acid could regulate the expression of P53,Bcl-2 and CASP3 mRNA and protein in Bel-7402 cells,and ursolic acid could promote the overexpression of P53 and CASP3 mRNA and protein and inhibit the expression of Bcl-2 mRNA and protein in Bel-7402 cells.Conclusion:(1)Network pharmacology prediction shows that ursolic acid,the main active component of AiTongXiao,may play a role in inhibiting hepatocellular carcinoma through P53 signaling pathway.(2)Ursolic acid significantly inhibited the proliferation of Bel-7402 cells in a dose-dependent manner,and activated P53 mediated Bcl-2/CASP3 mitochondrial apoptosis pathway to induce apoptosis.(3)AiTongXiao may inhibit hepatocellular carcinoma through P53/Bcl-2/CASP3 signaling pathway. |
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