TMT-Based Quantitative Proteomics Reveals the Targets of Andrographolide on LPS-induced Liver Injury

AbstractBackground Andrographolide is a diterpenoid derived from Andrographis paniculate, which has anti-inflammatory, antibacterial, antiviral and hepatoprotective activities. Gram-negative bacterial infections can cause varying degrees of liver injury in chickens, although andrographolide has been shown to have a protective effect on the liver, its underlying mechanism of action and effects on liver proteins are not known. Methods The toxicity of andrographolide on the viability of LMH cells at different concentrations and times was analyzed by CCK-8 assays. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in the culture supernatants were measured using an automatic biochemical analyzer to evaluate the protective effect of androscopolide on LPS-induced injury of LMH cells. Subsequently, TMT proteomics analysis were performed on the NC, LPS, and LPS-Andro groups, and bioinformatics analysis was performed on the differentially expressed proteins (DEPs). Results It was found that andrographolide reduced ALT and AST levels in the cell supernatant and alleviated LPS-induced injury in LMH cells. Proteomic analysis identified 50 and 166 differentially expressed proteins in the LPS vs NC group and LPS-Andro vs LPS group, respectively. Andrographolide may be involved in steroid metabolic processes, negative regulation of MAPK cascade, oxidative stress, and other processes to protect against LPS-induced liver injury. Conclusions Andrographolide protects against LPS-induced liver injury, HMGCS1, HMGCR, FDPS, PBK, CAV1, PRDX1, PRDX4, and PRDX6, which were identified by differential proteomics, may be the targets of andrographolide. Our study may provide new theoretical support for andrographolide protection against liver injury.