Sea buckthorn flavonoids attenuate oxidative stress in hepatocellular carcinoma via suppression of PI3K/Akt and JAK pathways.

OBJECTIVE: Sea buckthorn is rich in flavonoid components that possess significant antioxidant capacity, capable of eliminating free radicals, reducing oxidative damage, and demonstrating inhibitory effects on the proliferation of various cancer cell lines. The total flavonoids of sea buckthorn (SFs) and its monomers have shown their effect in inhibiting the proliferation of human liver cancer cells in vitro, accompanied by a decrease in intracellular oxidative stress indicators. Although current research results indicate that SFs have antioxidant and anti-hepatocellular carcinoma activities, there is still a lack of precise molecular mechanism studies on their role in Hepatocellular carcinoma (HCC).

METHODS: SFs were extracted using an ultrasonication-assisted ammonium sulfate-ethanol method and characterized by LC-MS. Network pharmacology identified core targets, which were subsequently validated through molecular docking and dynamics simulations. In vitro, the antioxidant capacity of SFs was assessed using 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), Peroxyl Radical Scavenging Capacity (PSC), and Total Antioxidant Capacity (TOAC) assays. The effects of SFs on HepG2 cell viability and apoptosis were evaluated using CCK-8 and flow cytometry assays, respectively. Oxidative stress-related factors were measured by ELISA. Western blot and qPCR were used to determine the effect of SFs on the PI3K/Akt and JAK signaling pathways.

RESULT: LC-MS identified 1,988 compounds, including key flavonoids such as chrysin, hesperetin, acacetin, and nobiletin. Network pharmacology highlighted PI3K and JAK as core targets. To reveal the relationship between SFs and core target, molecular docking was used to find binding affinities between these key flavonoids and the core targets, which was corroborated by stable molecular dynamics simulations. Furthermore, we found that SFs exerted their antioxidant, anti-apoptotic, and anti-inflammatory effects through the inhibition of the PI3K/Akt and JAK pathways.

CONCLUSION: This study demonstrates that SFs attenuate oxidative stress in HepG2 cells by scavenging free radicals, inhibiting pro-inflammatory cytokines, and modulating the PI3K/Akt and JAK pathways. These findings position SFs as a strong candidate for further preclinical development as an adjuvant therapy for HCC.