Journal of Isfahan Medical School

Journal of Isfahan Medical School

The Effects of Silibinin on Gene Expression of NOX1, NOX2, and the Production of Reactive Oxygen Species in TGF-Β-Treated Liver Stellate Cells

Document Type : Original Article (s)

Authors
Mojtaba Rashidi
Reza Afarin
Elham Shakerian
Shahla Asadizadeh
Samaneh Salehipour Bavarsad
Cellular and Molecular Research Center, Department of Clinical Biochemistry, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
10.48305/jims.v40.i664.0172
Abstract
Background: Liver fibrosis is a chronic disease caused by viral infections (such as hepatitis B and C viruses), alcohol abuse, and metabolic and genetic disorders that leads to excessive accumulation of extracellular matrix proteins, including collagen. The progression of liver fibrosis can leads to cirrhosis and liver cancer. In this study, the role of silibinin in the prevention of liver fibrosis progression was investigated.
Methods: LX2 cells were cultured in DMEM medium with 10% Fetal Bovine Serum (FBS). In the first stage, cells were treated with TGF-β at a concentration of 2 ng / ml (fibrotic group) for cell damage and fibrotic conditions for 24 hours, then at concentrations of 10, 20, 40, 60 and 80 μM of silibinin (The treatment groups) were treated for 24 hours and the mRNA expression of αSMA, collagen1α, NOX1 and NOX2 genes and the rate of intracellular reactive oxygen species (ROS) production were measured.
Findings: The results showed that the mRNA expression of αSMA, collagen1α, NOX1 and NOX2 genes at concentrations of 60 and 80 μM silibinin was significantly reduced compared to the TGF-β group. Also, the rate of intracellular ROS production at 60 and 80 μM concentrations of silibinin was significantly reduced compared to the TGF-β group (P < 0.05).
Conclusion: According to our study, silibinin inhibits the activation of Hepatic stellate cells (HSCs) by reducing the expression of genes involved in the progression of hepatic fibrosis and reduces liver damage caused by excessive production of collagen and reactive oxygen species in vitro. The findings from this study indicate that silybinin may be a potential therapeutic agent in the treatment of liver fibrosis.
Keywords
Liver fibrosis
Silibinin
Reactive oxygen species
Transforming growth factor beta
  1. Seki E, Schnabl B. Role of innate immunity and the microbiota in liver fibrosis: crosstalk between the liver and gut. J Physiol 2012; 590(3): 447-58.
  2. Carloni V, Luong TV, Rombouts K. Hepatic stellate cells and extracellular matrix in hepatocellular carcinoma: more complicated than ever. Liver Int 2014; 34(6): 834-43.
  3. Seki E, De Minicis S, Österreicher CH, Kluwe J, Osawa Y, Brenner DA, et al. TLR4 enhances TGF-β signaling and hepatic fibrosis. Nat Med 2007; 13(11): 1324-32.
  4. Liu C, Chen X, Yang L, Kisseleva T, Brenner DA, Seki E. Transcriptional repression of the transforming growth factor β (TGF-β) Pseudoreceptor BMP and activin membrane-bound inhibitor (BAMBI) by Nuclear Factor κB (NF-κB) p50 enhances TGF-β signaling in hepatic stellate cells. J Biol Chem2014; 289(10): 7082-91.
  5. Qu Y, Zhang Q, Cai X, Li F, Ma Z, Xu M, et al. Exosomes derived from miR‐181‐5p‐modified adipose‐derived mesenchymal stem cells prevent liver fibrosis via autophagy activation. J Cell Mol Med 2017; 21(10): 2491-502.
  6. Crosas-Molist E, Fabregat I. Role of NADPH oxidases in the redox biology of liver fibrosis. Redox Biol 2015; 6: 106-11.
  7. Tamayo C, Diamond S. Review of clinical trials evaluating safety and efficacy of milk thistle (Silybum marianum [L.] Gaertn.). Integr Cancer Ther 2007; 6(2): 146-57.
  8. Xu F, Yang J, Negishi H, Sun Y, Li D, Zhang X, et al. Silibinin decreases hepatic glucose production through the activation of gut–brain–liver axis in diabetic rats. Food Funct 2018; 9(9): 4926-35.
  9. Lv DD, Wang YJ, Wang ML, Chen EQ, Tao YC, Zhang DM, et al. Effect of silibinin capsules combined with lifestyle modification on hepatic steatosis in patients with chronic hepatitis B. Sci Rep 2021; 11(1): 655.
  10. Shi YF, Zhang Q, Cheung PY, Shi L, Fong CC, Zhang Y, et al. Effects of rhDecorin on TGF-β1 induced human hepatic stellate cells LX-2 activation. Biochim Biophys Acta 2006; 1760(11): 1587-95.
  11. Trappoliere M, Caligiuri A, Schmid M, Bertolani C, Failli P, Vizzutti F, et al. Silybin, a component of sylimarin, exerts anti-inflammatory and anti-fibrogenic effects on human hepatic stellate cells. J Hepatol 2009; 50(6): 1102-11.
  12. Reungoat E, Grigorov B, Zoulim F, Pécheur EI. Molecular crosstalk between the hepatitis C virus and the extracellular matrix in liver fibrogenesis and early carcinogenesis. Cancers (Basel) 2021; 13(9): 2270.
  13. Schuppan D, Ashfaq-Khan M, Yang AT, Kim YO. Liver fibrosis: Direct antifibrotic agents and targeted therapies. Matrix Biol 2018; 68-69: 435-51.
  14. Afarin R, Babaahmadi Rezaei H, Yaghouti SH, Taghvaei NM. The effect of cholesterol on the activation of TGF-β/Smad3C signaling pathway in hepatic stellate cells and its role in the progression of liver fibrogenesis. J Isfahan Med Sch 2021; 39(619): 212-8. [In Persian].
  15. Aydın MM, Akçalı KC. Liver fibrosis. Turk J Gastroenterol 2018; 29(1): 14-21.
  16. Lin L, Gong H, Li R, Huang J, Cai M, Lan T, et al. Nanodrug with ROS and pH dual‐sensitivity ameliorates liver fibrosis via multicellular regulation. Adv Sci (Weinh) 2020; 7(7): 1903138.
  17. Kim HH, Choi SE, Jeong WI. Oxidative stress and glutamate excretion in alcoholic steatosis: Metabolic synapse between hepatocyte and stellate cell. Clin Mol Hepatol 2020; 26(4): 697.
  18. Paik YH, Kim J, Aoyama T, De Minicis S, Bataller R, Brenner DA. Role of NADPH oxidases in liver fibrosis. Antioxid Redox Signal 2014; 20(17): 2854-72.
  19. Lan T, Kisseleva T, Brenner DA. Deficiency of NOX1 or NOX4 prevents liver inflammation and fibrosis in mice through inhibition of hepatic stellate cell activation. PloS One 2015; 10(7): e0129743.
  20. Liang S, Kisseleva T, Brenner DA. The role of NADPH oxidases (NOXs) in liver fibrosis and the activation of myofibroblasts. Front Physiol 2016;
    7: 17.
Journal of Isfahan Medical School
Volume 40, Issue 664
4th Week, May
May and June 2022
Pages 172-178

  • Receive Date 04 June 2022