Quercetin, the Ingredient of Xihuang Pills, Inhibits Hepatocellular Carcinoma by Regulating Autophagy and Macrophage Polarization

Abstract

Background:The key active component(s) in an anti-tumor preparation used in traditional Chinese medicine, Xihuang Pills, remains unclear. Methods: We used a network pharmacology analysis to construct a component-disease-target network diagram and used this to determine quercetin as a critical active ingredient in Xihuang Pills. Subsequently, human hepatocellular carcinoma (HCC) cell lines, H22 and HepG2 cells, were treated with quercetin, and BALB/c mice were injected with H22 cells and treated with different concentrations of quercetin. Tumor volume and weight were determined in these mice with and without quercetin administration. Immune and pro-inflammatory factors were measured using Enzyme Linked Immunosorbent Assay (ELISA). Macrophage polarization was assessed by western blot and flow cytometry. Finally, PD-L1, autophagy-related proteins, and the NF-κB pathway were also analyzed. Results: Quercetin could significantly inhibit the proliferation, migration, and invasion characteristics of HCC cells and promote apoptosis in a concentration-dependent manner in vitro. After quercetin treatment, tumor volume and weight significantly decreased in vivo. Granulocyte-macrophage and granulocyte colony-stimulating factor (GM-CSF and G-CSF, respectively) levels were blunted in response to quercetin, as well as the PD-L1 level. CD86+ cell ratio was increased, while the CD206+ cell ratio was decreased, suggesting that macrophages tend to undergo M1 polarization in response to quercetin. The expression of LC3 II/I was increased, while the expression of p62 was down-regulated. The pro-inflammatory factors TNF-α, IL-6, and IL-17A, as well as NF-κB signaling were suppressed in a quercetin concentration-dependent manner. Conclusions: Quercetin is a key ingredient of anti-HCC activity in Xihuang Pills by regulating macrophage polarization and promoting autophagy via the NF-κB pathway.

Integration of meta-analysis and network pharmacology analysis to investigate the pharmacological mechanisms of traditional Chinese medicine in the treatment of hepatocellular carcinoma

“…The top five active ingredients identified in our screening are MOL000098, MOL000422, MOL000449, MOL000006, and MOL000354, corresponding to quercetin, kaempferol, stigmasterol, luteolin, and isorhamnetin. Studies have shown that quercetin is involved in HCC autophagy (Wu et al, 2022), kaempferol inhibits HCC cell migration (Ju et al, 2021), luteolin enhances the efficacy of chemotherapy drugs (Xu et al, 2016), and isorhamnetin prevents liver fibrosis through oxidative stress (Yang et al, 2016). Funnel diagram of ORR.…”

Selected Flavonols Targeting Cell Death Pathways in Cancer Therapy: The Latest Achievements in Research on Apoptosis, Autophagy, Necroptosis, Pyroptosis, Ferroptosis, and Cuproptosis

“…Studies on the effect of QUE on hepatocellular carcinoma cells showed that LC3II/I increases with the concentration of the compound used, while p62 decreases. The authors suggest that the activation of the NF-κB pathway, through its impact on p62 levels, could stimulate autophagy, as indicated by the decreased levels of IκBα and phosphorylated p65 [ 118 ]. These results are confirmed by studies carried out on lung cancer cells, where the use of QUE reduces the expression of the p62 protein and increases GFP-LC3B in a dose-dependent manner [ 119 ].…”

Targeting Oxidative Stress with Polyphenols to Fight Liver Diseases

“…Similarly, another in vitro and in vitro study by Wu et al using H22, HepG2, and BALB/c mice showed that quercetin was able to inhibit HCC proliferation and migration and promote apoptosis in a dose-dependent manner. After treatment by gavage, quercetin produced a reduction in the volume and weight of liver tumors in BALB/c mice through macrophage polarization in coordination with the NF-κB pathway to modulate the autophagic process [ 165 ].…”