Lipopolysaccharide (LPS) triggers oxidative damage in sheep hepatocytes, linked to ferroptosis. Ferulic acid (FA) is known for its antioxidative properties, but its protective role against LPS via ferroptosis regulation was unclear. The objective of this research is to explore the protective role of FA in mitigating LPS-induced oxidative stress in sheep hepatocytes. The experimental setup consisted of three groups: a control group, an LPS group treated with 10 µg/mL of LPS, and FA group that received both 10 µg/mL of LPS and 750 µg/mL of FA. We found that FA treatment decreased in contents of MDA and LDH. Metabolomics revealed that LPS affected glycerophospholipid metabolism, unsaturated fatty acids biosynthesis, ferroptosis, and arachidonic acid metabolism mainly by reducing the level of PUFAs and LPC in the hepatocyte supernatant, while FA affected glutathione metabolism by increasing L-cysteine, L-ornithine, L-glutamic acid, and L-glutamine. Moreover, transcriptomics demonstrated that the comparison of LPS and control groups were mainly enriched in arachidonic acid metabolism, glycerophospholipid metabolism, and ferroptosis, the comparison of FA and LPS groups was mainly enriched in glutathione metabolism. The results further confirmed the findings in the metabolomics and transcriptomics analyses, showing that LPS treatment upregulated the mRNA expression of ACSL4, LPCAT3, ALOX15, STEAP3, GPX4, GCLC, and GCL in hepatocytes, while reducing GSH and GR levels. In contrast, FA intervention attenuated LPS-induced iron overload by decreasing Fe²⁺ accumulation and suppressing the mRNA expression of ACSL4, LPCAT3, STEAP3, and ALOX15. Furthermore, FA enhanced the expression of GPX4, GCLC, GCLM, and restored GSH content in LPS-exposed hepatocytes. The above results demonstrated that the protective effect of FA on LPS-induced oxidative damage in the sheep hepatocytes was achieved by activating the GSH-GPX4 pathway and inhibiting lipid metabolism-mediated ferroptosis.

Dietary regulation of intramuscular fat (IMF) deposition and fatty acid composition offers an effective strategy to enhance meat nutritional value. As phytogenic supplements rich in bioactive compounds, complex plant extracts (CPE) have demonstrated potential in improving meat quality through lipid metabolism modulation while ensuring food safety. In this study, we used 36 female sheep, approximately 4 months old and with a similar weight (29.92 ± 2.52 kg), to investigate the effects of CPE supplementation (80 mg/kg) on lipid metabolism. After the 75-day standardized feeding trial, the sheep were subjected to humane slaughter procedures and collected the Longissimus dorsi muscle (LDM) for further experimental process. The findings indicate that CPE significantly increased (p < 0.05) the IMF content (36%) and total fatty acids concentration (10,045.79 to 26,451.99 ug/g). Lipid metabolism in LDM was mainly affected by regulating phospholipids (six lipid subclasses were affected). The qRT-PCR analysis showed that differential expressed genes, PLA2G2D and PLA2G4E, associated with lipid metabolism were significantly reduced. CPE appears to modulate the fatty acids through sphingolipid, linolenic acid metabolism, and glycosphingolipid biosynthesis pathways. Thus, this study uncoded the mechanisms of CPE on fatty acid, providing critical evidence that CPE can regulate the meat quality of ruminants.

Fatty acid contributes notably to meat nutrition value. A previous study investigated how complex plant extracts (CPE) can improve the fatty acid composition of ruminants, but the molecular mechanism remains unknown. This study aimed to examine the effect of dietary CPE supplementation on sheep growth performance and fatty acid composition in back fat (BF), combining lipidomics and transcriptomics analyses to explain the underlying mechanisms. Thirty-six female sheep, weighing 29.92 ± 2.52 kg and of a similar age (~4 months old), were randomly assigned into two groups: one received a basal diet (CTRL group), and the other received the same diet supplemented with 80 mg/kg of CPE (CPE group) for 75 days. The results revealed that the values of carcass fat content (GR) in the CPE group were significantly increased (p = 0.008), and the composition of fatty acid was changed. Lipidomic analysis indicated that CPE modulated lipid metabolism by regulating the contents of lipid molecules such as phosphatidylcholine (PC), fatty acyls (FAs), cardiolipin (CL), and triglyceride (TAG). After the addition of CPE, the lipid metabolism of BF was regulated mainly by regulating the glycerophospholipid metabolism, TNF signaling pathway, and cytokine–cytokine receptor interaction signaling pathway. These results revealed that changes in fatty acids were affected by the added CPE and corresponding molecular changes, which may provide new insights on the molecular level for applying CPE in sheep to improve fatty acid composition.