Emodin Inhibitsg Hepatocellular Carcinoma Invasion and Migration by Regulating the HDAC4/NF-κB (p65)/CXCL12 Signaling Axis and Suppressing M2 Macrophage Polarization.

Abstract

This study aims to investigate how emodin, an active component of Polygonum multiflorum, regulates macrophage polarization through the HDAC4/NF-κB/CXCL12 pathway and impacts the progression of hepatocellular carcinoma (HCC). The inhibitory effect of emodin on M2 polarization was assessed by collecting conditioned cell culture medium from HepG2 cells and treating macrophages with emodin. Network pharmacology and other experiments were used to identify HDAC4 as a key target, and this was further validated through molecular docking, surface plasmon resonance (SPR), and Western blot analysis. Additional validation of the HDAC4/NF-κB(p65)/CXCL12 axis regarding HCC progression was conducted using single-cell RNA sequencing and an in vivo HCC model. The results demonstrate that macrophages with HepG2 supernatant after treating, including emodin, tetrahydroxy stilbene glucoside and physcion, were added to reduce macrophage M2 polarization and inhibit HepG2 invasion and migration. Network pharmacology and protein analysis were used to discover the role of HDAC4 as the target of three drugs in HCC. The feasibility of emodin as a drug target for HDAC4 was explored through molecular docking simulation and SPR, and then verified through Western blot and nuclear plasma separation analysis. Likewise, the HDAC4/NF-κB (p65) pathway reduces M2 polarization and inhibits the invasion and migration of HepG2. By using a single-cell sequencing database to predict the high expression of chemokines in HCC, and verifying the effect of emodin's ability to reduce M2 polarization and inhibit HepG2 invasion and migration via the HDAC4/NF-κB (p65)/CXCL12 pathway, the results were likewise validated in vivo. This study elucidates the novel role of emodin in HCC progression and macrophage polarization, and demonstrates how emodin-mediated HDAC4 inhibition effectively attenuates HCC invasion and migration.