Blockade of GPX4-mediated ferroptosis underlies the protective effect of Forsythiae Fructus water extract against poly(I:C)-induced inflammation.
Guo X, Chen B, Su Z, Cao X, Dai L
Summary
7.8/10A traditional Chinese medicine made from forsythia fruits reduces inflammation by preventing a specific type of cell death called ferroptosis through the activation of a protective protein (GPX4). This research validates the plant's anti-inflammatory properties using modern molecular techniques and identifies wogonin as its active ingredient, potentially leading to new treatments for inflammation-related diseases.
Key Findings
Forsythiae Fructus water extract (LQKL) reduced pro-inflammatory cytokines IL-6 and TNF-α while suppressing MDA content and ROS production in both cell and zebrafish models
GPX4 knockdown substantially diminished LQKL's anti-inflammatory effects, establishing GPX4 activation as essential to the protective mechanism
Wogonin was identified as the key bioactive component in LQKL that directly binds GPX4 and mediates the anti-inflammatory response
Original Abstract
Forsythiae Fructus is a widely used traditional Chinese medicine commonly employed in the treatment of inflammation-related diseases. However, its underlying anti-inflammatory mechanisms remain incompletely elucidated. This study aimed to evaluate the therapeutic effects of Forsythiae Fructus water extract (LQKL) against poly(I:C)-induced inflammation and to explore whether it acts by inhibiting ferroptosis. The protective effects of LQKL were evaluated using poly(I:C)-induced inflammatory models in both RAW264.7 cells and zebrafish. Potential targets and pathways of LQKL against poly(I:C)-induced inflammation were initially predicted by network pharmacology analysis and subsequently validated through RT-qPCR, Western blot, immunofluorescence, siRNA transfection, and CRISPRi. Lipid peroxidation levels were assessed by measuring MDA content and ROS production using colorimetric assay and fluorescence microscopy, respectively. Furthermore, the key anti-inflammatory components in LQKL were identified through integrated approaches including molecular docking, DARTS, and CETSA assays. LQKL significantly attenuated poly(I:C)-induced inflammation in zebrafish, as evidenced by reduced neutrophil and macrophage infiltration and improved survival rate. It also downregulated the expressions of IL-6 and TNF-α in both poly(I:C)-stimulated zebrafish and RAW264.7 cells. Network pharmacology analysis indicated that LQKL may act through inhibition of the NF-κB signaling pathway which were verified by the decrease of p-NF-κB p65 and p-IκBα, as well as the nuclear translocation of NF-κB p65. Moreover, LQKL inhibited MDA content, suppressed the generation of ROS, restored GSH content and regulated the expressions of proteins related to ferroptosis. Notably, GPX4 knockdown markedly diminished the anti-inflammatory effects of LQKL and its suppression of the NF-κB pathway. Furthermore, wogonin was identified as a key active component in LQKL that strongly binds to GPX4. LQKL alleviates poly(I:C)-induced inflammation by attenuating ferroptosis through GPX4 activation, leading to subsequent inhibition of the NF-κB pathway.
Species Mentioned
Forsythia, is a genus of flowering plants in the olive family Oleaceae. There are about 11 species, mostly native to Eastern Asia, but one native to Southeastern Europe. Forsythia – also one of the plant's common names – is named after the botanist William Forsyth.
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