Total flavonoids of Salvia miltiorrhiza alleviate acetaminophen-induced acute liver injury in mice by suppressing hepatocyte ferroptosis via activating the Nrf2/HO-1 signaling pathway

doi:10.12122/j.issn.1673-4254.2024.11.17

Abstract

Abstract:

Objective To investigate the protective effect of total flavonoids of Salvia divinorum extract against acetaminophen (APAP)-induced acute liver injury (ALI) and its molecular mechanism. Methods The main chemical constituents of total flavonoids of Salvia divinorum were obtained through literature search, and their pharmacological mechanisms were predicted using bioinformatics analysis. In a mouse model of APAP-induced ALI, the protective effects of 100, 200 and 400 mg/kg total flavonoids of Salvia miltiorrhiza and 150 mg/kg bifidus were evaluated by observing changes in blood biochemistry and liver histopathology and detecting expressions of the key proteins in the Nrf2/HO-1 signaling pathway. Results Network pharmacology analysis suggested that the main active components in total flavonoids of Salvia divinorum for regulating APAP-induced liver injury included quercetin, lignocerol, caruric acid, and kaempferol, for which GO function enrichment analysis yielded 632 GO entries, including 472 involving biological processes, 42 involving cellular composition, and 118 involving molecular function. KEGG enrichment analysis showed that the total flavonoids of Salvia divinorum regulated APAP-induced liver injury mainly through ferroptosis-related signaling pathway. In mice with APAP-induced ALI, treatment with the total flavonoids significantly lowered ALT and AST levels, improved liver histopathology and inflammatory cell infiltration, reduced iron deposition in liver tissues, improved lipid peroxidation-related indexes, promoted the expressions of Nrf2, HO-1, SLC7A11, and GPX-4 proteins, and inhibited the expression of keap1 protein. Conclusion The total flavonoids of Salvia divinorum alleviate APAP-induced ALI in mice possibly by suppressing hepatocyte ferroptosis via activating the Nrf2/SLC7A11/GPX-4 signaling pathway.

Key words: total flavonoids of Salvia divinorum, Nrf2/HO-1, ferroptosis, network pharmacology, acute liver injury

Huajun CAI, Zhiqi CHEN, Wenting HU, Wei TAN, Hao WU, Chao WANG. Total flavonoids of Salvia miltiorrhiza alleviate acetaminophen-induced acute liver injury in mice by suppressing hepatocyte ferroptosis via activating the Nrf2/HO-1 signaling pathway[J]. Journal of Southern Medical University, 2024, 44(11): 2201-2208.

Figures/Tables 8

Fig.1 Result of network pharmacology analysis. A: Intersection target Wayne plot of the ethanol extract of Herba Desmodii Styracifolii and liver injury caused by acetaminophen. B: Protein-protein interaction (PPI) network. C: Drug-active ingredient-target-disease network.

Fig.2 GO analysis ( A) and KEGG enrichment analysis diagram ( B).

Fig.3 Effect of total flavonoids of Salvia divinorum extract (TFM) on serum ALT and AST levels in mice with APAP-induced ALI ( Mean±SD, n=6). ** P<0.01 vs control group; #P<0.05, ##P<0.01 vs APAP group.

Fig.4 Pathological changes in the liver tissues of mice with APAP-induced ALI treated with TFM (HE staining).

Fig.5 Effect of TFM on serum inflammatory factor TNF- α and IL-6 in the mice. ** P<0.01 vs control group; #P<0.05, ##P<0.01 vs APAP group.

Fig.6 Effects of TFM on the contents of 4-HNE ( A), MDA ( B), ROS ( C), and GSH( D) in the liver tissues of the mice. ** P<0.01 vs control group; #P<0.05, ##P<0.01 vs APAP group.

Fig.7 TFM reduces iron deposition in the liver tissues of the mice. A: Perls staining; B: Quantitative results ofPerls staining. ** P<0.01 vs control group; #P<0.05, ##P<0.01 vs APAP group.

Fig.8 　Expression levels of Nrf2, HO-1, keap1, GPX-4 and SLC7A11 proteins in the liver tissues of the mice detected by Western blotting. **P<0.01vs control group; #P<0.05, ##P<0.01vs APAP group.

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