From Bench to Bedside: Eupatilin Activates Antioxidant Defenses and Reduces Fibrosis in Experimental Cholestasis

Abstract

Aim

The aim of this study is to explore the protective effects and mechanisms of Eupatilin, a peroxisome proliferator-activated receptor α (PPARα) agonist, on cholestatic liver disease induced by common bile duct ligation (BDL) in mice.

Materials and Methods

We selected Balb/c mice (both male and female) aged 6 to 8 weeks for the common BDL procedure (ethical approval number: MUST-FDCT-20241114001). The groups include the BDL group and the BDL+ Eupatilin group, with three mice in each group. Once the mice developed jaundice postsurgery (5 days), they were treated with Eupatilin via gavage at a dosage of 20 mg/kg daily for a period of 8 days. On day 13, ocular blood was collected, and liver tissues were extracted for histopathological examination with H&E staining, Sirius Red staining, and subsequent RNA sequencing. Statistical differences among the parameters were evaluated using a t-test.

Main Results

Eupatilin reduces the liver weight/body weight ratio by 41% and ameliorates liver necrosis and fibrosis in Balb/c mice. It could decrease alanine transaminase ( p = 0.0498), aspartate aminotransferase (p = 0.0077), while maintaining ALB (Albumin) and γ-GT (gamma-glutamyl transferase) within normal ranges. RNA sequencing analysis revealed that antioxidant genes (acetaldehyde dehydrogenase 2 [Aldh2] and superoxide dismutase 1 [Sod1]) might be the targets of Eupatilin action.

Conclusion

We found that Eupatilin can upregulate antioxidant genes (Aldh2; p = 0.0107) and Sod1 (p = 0.0208) of Balb/c mice, thereby ameliorating BDL damage in mice with cholestatic liver disease.

Keywords

Eupatilin - bile duct ligation - antioxidant - cholestatic liver disease

Publikationsverlauf

Eingereicht: 12. April 2025

Angenommen: 03. August 2025

Artikel online veröffentlicht:
20. August 2025

• References

• 1 Li J, Zhu X, Zhang M. et al. Limb expression 1-like (LIX1L) protein promotes cholestatic liver injury by regulating bile acid metabolism. J Hepatol 2021; 75 (02) 400-413
• 2 Han D, Kim H, Kim S. et al. Sestrin2 protects against cholestatic liver injury by inhibiting endoplasmic reticulum stress and NLRP3 inflammasome-mediated pyroptosis. Exp Mol Med 2022; 54 (03) 239-251
• 3 Huang W, Qian Y, Lin J, Wang F, Kong X, Tan W. Baicalein alleviates intrahepatic cholestasis by regulating bile acid metabolism via an FXR-dependent manner. Biochem Biophys Res Commun 2024; 705: 149670
• 4 Xi XJ, Chen SH, Mi H. Aldh2 gene reduces oxidative stress in the bladder by regulating the NF-κB pathway in a mouse model of ketamine-induced cystitis. Exp Ther Med 2020; 20 (05) 111
• 5 Wang MQ, Zhang KH, Liu FL. et al. Wedelolactone alleviates cholestatic liver injury by regulating FXR-bile acid-NF-κB/NRF2 axis to reduce bile acid accumulation and its subsequent inflammation and oxidative stress. Phytomedicine 2024; 122: 155124
• 6 Baek J, Lee MG. Oxidative stress and antioxidant strategies in dermatology. Redox Rep 2016; 21 (04) 164-169
• 7 Ge W, Guo R, Ren J. AMP-dependent kinase and autophagic flux are involved in aldehyde dehydrogenase-2-induced protection against cardiac toxicity of ethanol. Free Radic Biol Med 2011; 51 (09) 1736-1748
• 8 Atiya A, Muhsinah AB, Alrouji M. et al. Unveiling promising inhibitors of superoxide dismutase 1 (SOD1) for therapeutic interventions. Int J Biol Macromol 2023; 253 (Pt 2): 126684
• 9 Lepore AC. Intraspinal cell transplantation for targeting cervical ventral horn in amyotrophic lateral sclerosis and traumatic spinal cord injury. J Vis Exp 2011; (55) 3069
• 10 Park SC, Yoon JH, Kim W. et al. Eupatilin attenuates bile acid-induced hepatocyte apoptosis. J Gastroenterol 2006; 41 (08) 772-778
• 11 Nene A, Chen CH, Disatnik MH, Cruz L, Mochly-Rosen D. Aldehyde dehydrogenase 2 activation and coevolution of its εPKC-mediated phosphorylation sites. J Biomed Sci 2017; 24 (01) 3
• 12 Wang Y, Branicky R, Noë A, Hekimi S. Superoxide dismutases: Dual roles in controlling ROS damage and regulating ROS signaling. J Cell Biol 2018; 217 (06) 1915-1928
• 13 Kim JS, Cha KH, Kang SY. et al. In vivo gastric residence and gastroprotective effect of floating gastroretentive tablet of DA-9601, an extract of Artemisia asiatica, in beagle dogs. Drug Des Devel Ther 2016; 10: 1917-1925
• 14 Li Z, Wang X, Tasich K. et al. Eupatilin unveiled: An in-depth exploration of research advancements and clinical therapeutic prospects. J Transl Int Med 2025; 13 (02) 104-117
• 15 Jung Y, Kim JC, Choi Y. et al. Eupatilin with PPARα agonistic effects inhibits TNFα-induced MMP signaling in HaCaT cells. Biochem Biophys Res Commun 2017; 493 (01) 220-226
• 16 Jung Y, Kim JC, Park NJ. et al. Eupatilin, an activator of PPARα, inhibits the development of oxazolone-induced atopic dermatitis symptoms in Balb/c mice. Biochem Biophys Res Commun 2018; 496 (02) 508-514
• 17 Pawlak M, Lefebvre P, Staels B. Molecular mechanism of PPARα action and its impact on lipid metabolism, inflammation and fibrosis in non-alcoholic fatty liver disease. J Hepatol 2015; 62 (03) 720-733
• 18 Moustafa T, Fickert P, Magnes C. et al. Alterations in lipid metabolism mediate inflammation, fibrosis, and proliferation in a mouse model of chronic cholestatic liver injury. Gastroenterology 2012; 142 (01) 140-151.e12
• 19 Trivella J, John BV, Levy C. Primary biliary cholangitis: Epidemiology, prognosis, and treatment. Hepatol Commun 2023; 7 (06) e0179
• 20 Thanh Loc V, Bolor T, Purevdorj E. et al. Therapeutic effects of Chrysanthemum zawadzkii and its active compound eupatilin in rheumatoid arthritis: in silico and in vitro validation. Nat Prod Res 2025; 1-8
• 21 Liu S, Li T, Yang Q, Ke X, Zhan J. Biliary atresia: the development, pathological features, and classification of the bile duct. Pediatr Surg Int 2024; 40 (01) 42
• 22 Pal N, Joy PS, Sergi CM. Biliary atresia animal models: Is the needle in a haystack?. Int J Mol Sci 2022; 23 (14) 7838