Research Paper Volume 15, Issue 21 pp 11994—12020
Hepatoprotective effect of syringin combined with costunolide against LPS-induced acute liver injury in L-02 cells via Rac1/AKT/NF-κB signaling pathway
- 1 Chongqing Medical and Pharmaceutical College, Chongqing 400030, China
- 2 College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, China
- 3 Chongqing Key Laboratory of High Active Traditional Chinese Drug Delivery System, Chongqing 400030, China
- 4 Chongqing Three Gorges Medical College, Chongqing 404120, China
Received: August 10, 2023 Accepted: October 2, 2023 Published: November 1, 2023
https://doi.org/10.18632/aging.205161How to Cite
Copyright: © 2023 Mao et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Acute liver injury (ALI) leads to abnormal liver function and damage to liver cells. Syringin (syr) and costunolide (cos) are the major extracts from Dolomiaea souliei (Franch.) C.Shih (D. souliei), showing diverse biological functions in various biological processes. We explored the underlying hepatoprotective effects of syr+cos against LPS-induced ALI. Cell viability and proliferation were assessed using an MTT assay and immunofluorescence staining. Flow cytometry analysis was used to detect cell cycle distribution and apoptosis. ELISA was utilized to measure liver function and antioxidant stress indexes. qRT-PCR and western blotting was performed to determine mRNA and protein levels respectively. Using shRNA approach to Rac1 analyzed transcriptional targets. The results showed that syr+cos promoted L-02 cell proliferation, inhibiting the cell apoptosis and blocking cell cycle in G1 and G2/M phase. Syr+cos decreased the production of ALT, AST, LDH, MDA and ROS while increased SOD and CAT activities. Pretreated with syr+cos may decrease expressions of caspase-3,7,9, NF-κB, TNF-α proteins, Cyclin B, CDK1 and p-IκB proteins while p-IκB increased. Silencing of Rac-1 may protect the liver by increasing AKT, S473, T308 and reducing p-AKT proteins. Syr+cos exhibits anti-ALI activity via Rac1/AKT/NF-κB signaling pathway which might act as an effective candidate drug for the treatment of ALI.
Abbreviations
ALI: Acute liver injury; ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; Bax: Bcl-2 associated X protein; Bcl-2: B-cell lymphoma-2 protein; Caspase-3,7,9: Cysteinyl aspartate specific proteinase 3,7,9; CAT: Catalase; CDK1: Cyclin-dependent kinase 1; Cyclin B: Cyclin protein B; DAPI: 4’,6-diamidino-2-phenylindole; DCFH-DA/H2DCFDA: 2’,7’-dichlorofluorescein diacetate; DCF: 2’,7’-dichlorofluorescein; DMSO: Dimethyl sulfoxide; D. souliei: Dolomiaea souliei (Franch.) C.Shih; EDTA: ethylene diamine tetraacetie acid; ELISA: Enzyme linked immunosorbent assay; LDH: Lactic dehydrogenase; LPS: Lipopolysaccharide; MDA: Malondialdehyde; MTT: 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide; syr: Syringin; shRNA: short hairpin RNA; NF-κB: Nuclear factor-kappa B; ROS: Reactive oxygen species; RT-PCR: Reverse transcription polymerase chain reaction; SOD: Superoxide dismutase; Sily: Silymarin; PBS: Phosphate-buffered saline; PCR: Polymerase chain reaction; TNF-α: Tumor necrosis factor-α.