Exploring the Effect of Dan Zhi Xiao Yao San Jia Wei on Hepatocellular Carcinoma Cells Based on PI3K/Akt/FOXO Signa-ling Pathway

GU Xiufeng; LONG Zhixiong; ZHANG Dezhong; HAO Yaming

doi:10.13730/j.issn.2097-2148.2025.09.001

Abstract：Objective To investigate the mechanism of Dan Zhi Xiao Yao San Jia Wei intervening in primary hepatocellu-lar carcinoma by regulating phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/forkhead box(FOXO)signaling path-way.Methods A total of 5 SPF male SD rats aged 6 to 8 weeks were gavaged with 2 times Dan Zhi Xiao Yao San Jia Wei dosage(39.98 g/kg)according to the body surface area for 3 d,1 h after the last administration,blood was collected from the abdominal aorta,after centrifugation,the superna-tant was collected as drug-containing serum,filtered and ster-ilized for use.HepG2 hepatocellular carcinoma cells were ran-domly divided into control group and experimental group,HepG2 hepatocellular carcinoma cells in the experimental group were treated with serum containing 5％,10％,15％and 20％Dan Zhi Xiao Yao San Jia Wei,respectively,the control group(0％serum containing medicine)was routinely cultured.Proliferation inhibition rate was detected by cell counting kit-8(CCK-8);cell apoptosis rate and proliferation rate were detected by flow cytometry;scratch test was used to detect cell migration ability;Tran-swell assay was used to detect cell invasion ability;the expressions of PI3K,Akt and FOXO1 mRNA were detected by reverse transcription polymerase chain reaction(RT-PCR);Western blot was used to detect PI3K,Akt,FOXO1 and FOXO downstream apoptosis-related proteins B-cell lymphoma 2(Bcl-2)and Bcl-2 associated X protein(Bax)protein expressions.Results ① Com-pared with the control group,the OD of 10％,15％and 20％drug-containing serum in the experimental groups decreased,the cell proliferation inhibition rate increased(all P＜0.01);in the experimental group,with the increase of drug-containing serum concentration,OD decreased gradually,and cell proliferation inhibition rate increased significantly(all P＜0.05).② Compared with the control group,the apoptosis rate of cells in the experimental groups with 10％,15％and 20％drug-containing serum increased,and the cells proliferation rate,migration distance,invasive number of cells and the invasive number of cells in the ex-perimental groups with 5％drug-containing serum decreased(all P＜0.05);in the experimental group,with the increase of drug-containing serum concentration,the apoptosis rate increased,and the cell proliferation rate,migration distance and invasion number decreased(all P＜0.05).③ Compared with the control group,the expression levels of PI3K,Akt and FOXO1 mRNA in the experimental group decreased,and the differences were statistically significant(all P＜0.05),except for Akt mRNA level in 5％drug-containing serum(P＞0.05);in the experimental group,with the increase of drug-containing serum concentration,the inhibition trend was gradually significant.④ Compared with the control group,the levels of PI3K,Akt,FOXO1 and Bcl-2 protein in the experimental group decreased,and the level of Bax protein increased(all P＜0.01),except for the level of FOXO1 protein in the 5％drug-containing serum(P＞0.05);with the increase of drug-containing serum concentration,the trend was more significant.Conclusion Dan Zhi Xiao Yao San Jia Wei can significantly inhibit the proliferation,migration and invasion of HepG2 hepatocellular carcinoma cells,induce cell apoptosis,and its mechanism may be related to the regulation of PI3K/Akt/FOXO signaling pathway.