Tangeretin alleviates LPS-induced acute lung injury by inhibiting ferroptosis of macrophage via Nrf2 pathways
Sepsis-induced acute lung injury (ALI) is a severe clinical condition with a high mortality rate. Tangeretin, widely found in citrus fruits, has been reported to exert antioxidant and anti-inflammatory properties. However, whether Tangeretin protects against sepsis-induced ALI and the potential mechanisms remain unclear.We established ALI model via intraperitoneally injected with 5 mg/kg lipopolysaccharides (LPS) for 12 h. Tangeretin was applied intraperitoneally 30 min before LPS treatment.  The lung tissue samples from both the LPS and LPS + TAN groups were subjected to RNA sequencing analysis, conducted by OE Biotech Co., Ltd. (Shanghai, China). We performed differential gene expression analysis using RNA-seq data between LPS and LPS/Tangeretin group.GSEA analysis between LPS and LPS/Tangeretin group showed that IL6_JAK_STAT3_SIGNALING, INFLAMMATORY_RESPONSE, TNFA_SIGNALING_VIA_NFKB were significantly enriched (Fig.3C-E). These results identified the anti-inflammatory effect of Tangeretin against sepsis-induced ALI. Overall design: To induce a septic lung injury model, an intraperitoneal injection of lipopolysaccharide (LPS) extracted from Escherichia coli O111:B4 (L2630, Sigma-Aldrich, USA) was administered at a dosage of 5 mg/kg. Preceding this, tangeretin (Tan, HY-N0133, MCE corporation, USA) was given at 50 mg/kg intraperitoneally, 30 min prior to the LPS injection. The lung tissue samples from both the LPS and LPS + TAN groups were subjected to RNA sequencing analysis, conducted by OE Biotech Co., Ltd. (Shanghai, China). Briefly, Total RNA was extracted using TRIzol reagent (Invitrogen, USA), followed by an evaluation of its purity and quantification. RNA integrity was assessed via the Agilent 2100 Bioanalyzer (Agilent Technologies, USA). Subsequently, cDNA libraries were constructed using VAHTS Universal V6 RNA-seq Library Prep Kit, and the sequencing was executed on Illumina Novaseq 6000 platform. Approximately 50 M clean reads per sample were retained.