Study on the Mechanism of Shentao Ruangan Formula Inhibiting Cell Viability of Human Hepatoma Cells HepG2 / 世界科学技术-中医药现代化

ABSTRACT

Objective To explore the effects and mechanism of Shentao Ruangan Prescription on hepatocellular carcinoma.Methods MTT assay was used to detect the activity of Shentao Ruangan Prescription against HepG2 cells and the effects of PI3K/Akt/MAPK inhibitors(LY294002,Akti,MEKi,JNKi),apoptosis inhibitor(Z-VAD)and autophagy inhibitor(Wortmanin)in human hepatoma cells.),ferroptosis inhibitor(Ferrostatin-1)and cell necrosis inhibitor(Necrostatin-1)on the inhibition of HepG2 activity of human hepatoma cells by Shentao Ruangan Prescription.LDH method was used to detect the toxicity of Shentao Ruangan Prescription on HepG2 cells;Real-time PCR experiment tested the effect of Shentao Ruangan Prescription on the pyroptosis marker IL-1β in human hepatoma cells HepG2;Western blot experiment explored the effect of Shentao Ruangan Prescription on the Akt/ERK signaling pathway in human hepatoma HepG2 cells and the expression of Akt,ERK,P-ERK,the key proteins of apoptosis signaling pathway Caspase 3,PARP,Bcl2,Bax,and the expression of pyroptosis marker Cleavaged Caspase-1.Results Shentao Ruangan Prescription Can effectively inhibit the activity of human hepatoma HepG2 cells.LY294002,Wortmanin,Akti,MEKi,JNKi,Z-Vad,Ferrostatin-1,Necrostatin-1 had no significant effect on the inhibition of HepG2 activity by total extract(water extract),supernatant and precipitation.Shentao Ruangan Prescription promoted the release of LDH in human hepatoma HepG2 cells,causing cytotoxicity.It inhibited HepG2 pyroptosis markers in human hepatoma cells and the expression of IL-1β and Cleavaged Caspase-1 and decreased the protein expression of Akt in human hepatoma cells HepG2 but inversely increased Caspase3 protein expression.Conclusion Shentao Ruangan Prescription has the effect of inhibiting the activity of hepatocellular carcinoma cells and anti-hepatocellular carcinoma,and its mechanism may be related to inhibiting the expression of Akt and promoting the expression of Caspase3.At the same time,Shentao Ruangan Prescription reduces the expressions of IL-1β and Cleavaged Caspase-1,which may inhibit cell pyroptosis.The inhibitory effect of Shentao Ruangan Prescription on the activity of liver cancer cells is likely to be independent of programmed death modes such as autophagy and necrotizing apoptosis,and PI3K/Akt/MAPK signal transduction.The total extract of Shentao Ruangan Prescription has better anti-hepatoma activity than its water-soluble and alcohol-soluble components,and its water-soluble and alcohol-soluble components may have synergistic effect.