The Journal of Internal Korean Medicine (대한한방내과학회지)

The Society of Internal Korean Medicine (대한한방내과학회)
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Liver Protective Effect of the Co-treatment of Rhei Radix et Rhizoma and Silymarin on TAA-induced Liver Injury

대황과 실리마린의 병용투여의 간섬유화 보호 효과

  • Il-ha Jeong (Dept. of Herbology, College of Korean Medicine, Daegu Haany University) ;
  • Sang-woo Ji (Dept. of Herbology, College of Korean Medicine, Daegu Haany University) ;
  • Seong-soo Roh (Dept. of Herbology, College of Korean Medicine, Daegu Haany University)
  • 정일하 (대구한의대학교 한의과대학 본초약리학교실) ;
  • 지상우 (대구한의대학교 한의과대학 본초약리학교실) ;
  • 노성수 (대구한의대학교 한의과대학 본초약리학교실)
  • Received : 2023.05.22
  • Accepted : 2023.06.26
  • Published : 2023.06.30
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Abstract

Objective: Liver fibrosis is a highly conserved wound-healing response and the final common pathway of chronic inflammatory injury. This study aimed to evaluate the potential anti-fibrotic effect of the combination of Rhei Radix et Rhizoma water extract (RW) and silymarin in a thioacetamide (TAA)-induced liver fibrosis model. Methods: The liver fibrosis mouse model was established through the intraperitoneal injection of TAA (1 week 100 mg/kg, 2-3 weeks 200 mg/kg, 4-8 weeks 400 mg/kg) three times per week for eight weeks. Animal experiments were conducted in five groups; Normal, Control (TAA-induced liver fibrosis mice), Sily (silymarin 50 mg/kg), RSL (RW 50 mg/kg+silymarin 50 mg/kg), and RSH (RW 100 mg/kg+silymarin 50 mg/kg). Biochemical analyses were measured in serum, including aspartate aminotransferase (AST), alanine aminotransferase (ALT), malondialdehyde (MDA), and ammonia levels. Liver inflammatory cytokines and fibrous biomarkers were measured by Western blot analysis, and liver histopathology was evaluated through tissue staining. Results: A significant decrease in the liver function markers AST and ALT and a reduction in ammonia and total bilirubin were observed in the group treated with RSL and RSH. Measurement of reactive oxygen species and MDA revealed a significant decrease in the RSL and RSH administration group compared to the TAA induction group. The expression of extracellular matrix-related proteins, such as transforming growth factor β1, α-smooth muscle actin, and collagen type I alpha 1, was likewise significantly decreased. All drug-administered groups had increased matrix metalloproteinase-9 but a decreasing tissue inhibitor of matrix metalloproteinase-1. RSL and RSH exerted a significant upregulation of NADPH oxidase 2, p22phox, and p47phox, which are oxidative stress-related factors. Furthermore, pro-inflammatory proteins such as cyclooxygenase 2 and interleukin-1β were markedly suppressed through the inhibition of nuclear factor kappa B activation. Conclusions: The administration of RW and silymarin suppressed the NADPH oxidase factor protein level and showed a tendency to reduce inflammation-related enzymes. These results suggest that the combined administration of RW and silymarin improves acute liver injury induced by TAA.

Keywords

Acknowledgement

본 연구는 2022년도 정부(과학기술정보통신부)의 재원으로 한국연구재단의 지원(No. 2018R1A5A2025272)을 받아 수행되었습니다. 또한 과학기술정보통신부(MSIT)의 R&D 복귀자 프로그램에 따라 한국과학기술여성재단(WISET) 보조금의 지원(2021-014)을 받았습니다.

References

  1. Kisseleva T, Brenner DA. Role of hepatic stellate cells in fibrogenesis and the reversal of fibrosis. J Gastroenterology and Hepatology 2007;22(1):78-88.  https://doi.org/10.1111/j.1440-1746.2006.04658.x
  2. Ramos-Tovar E, Muriel P. Molecular Mechanisms that link oxidative stress, inflammation, and fibrosis in the liver. Antioxidants 2020;9(12):1249. 
  3. Xu J, Liu X, Koyama Y, Wang P, Lan T, Kim IG, et al. The type of hepatic myofibroblasts contributing to liver fibrosis of different etiologies. Front Pharmacol 2014;5:167. 
  4. Ozdemir-Kumral ZN, Erkek BE, Karakus B, Almaci M, Fathi R, Yuksel M, et al. Potential effect of 1,25 Dihydroxyvitamin D3 on thioacetamide-induced hepatotoxicity in rats. J Surg Res 2019;243:165-72.  https://doi.org/10.1016/j.jss.2019.05.020
  5. Lee JA, Lee SH, Shin MR, Sook NJ, Roh SS. The Effect of Chaenomelis Fructus Extract on Thioacetamide-induced Fibrosis. JKSFSN 2021;50(4):322-9.  https://doi.org/10.3746/jkfn.2021.50.4.322
  6. Muller A, Machnik F, Zimmermann T, Schibert H. Thioacetamide-induced cirrhosis-like liver lesions in rats-usefulness and reliability of this animal model. Exp Pathol 1998;34(4):229-36.  https://doi.org/10.1016/S0232-1513(88)80155-5
  7. Higashi T, Friedman SL, Hoshida Y. Hepatic stellate cells as key target in liver fibrosis. Adv Drug Deliv Rev 2017;121:27-42.  https://doi.org/10.1016/j.addr.2017.05.007
  8. Breitkopf K, Godoy P, Ciuclan L, Singer MV, Dooley S. TGF-beta/Smad signaling in the injured liver. Z Gastroenterol 2006;44(1):57-66.  https://doi.org/10.1055/s-2005-858989
  9. Hassan HA. Oxidative stress as a crucial factor in liver associated disorders: Potential therapeutic effect of antioxidants. The Liver, Academic Press 2018:121-30. 
  10. Lieber CS, Leo MA, Cao Q, Ren Cm Decarli LM. Silymarin retards the progression of alcohol-induced hepatic fibrosis in baboons. J Clin Gastroenterol 2003;37(4):336-9.  https://doi.org/10.1097/00004836-200310000-00013
  11. Jeong JH, Kim JH, Lee JD. A review on the medicinal potentials of Radix et Rhizoma Rhei. Journal of pharmacopuncture 2015;18(3):7-16. 
  12. Kiruthga PV, Shafreen RB, Pandian SK, Devi KP. Silymarin protection against major reactive oxygen species released by environmental toxins: exogenous H2O2 exposure in erythrocytes. Basic Clin Pharmacol Toxicol 2007;100(6):414-9.  https://doi.org/10.1111/j.1742-7843.2007.00069.x
  13. Di Costanzo A, Angelico R. Formulation strategies for Enhancing the Bioavailability of Silymarin: The state of the art. Molecules 2019;24(11):2155. 
  14. Song JH, Yang TC, Chang KW, Han SK, Yi HK, Jeon JG. In vitro anti-cariogenic activity of dichloromethane fraction from Rheum undulatum L. root. Archives of pharmacal research 2006;29(6):490-6.  https://doi.org/10.1007/BF02969422
  15. Hwang IY, Jeong CS. A study on the Antigastric effects of Rheum Species Extracts and Their Active Components. Journal of food Hygiene and safety 2013;2(4):330-6.  https://doi.org/10.13103/JFHS.2013.28.4.330
  16. Folin O, Denis W. On phosphotungasticphosphomolybdic compounds as color reagent. Journal of biological chemisty 1912;12(2):239-43.  https://doi.org/10.1016/S0021-9258(18)88697-5
  17. Lister CE, Lancaster JE, Sutton KH, Walker JR. Developmental changes in the concentration and composition of flavonoids in skin of a red and a green apple cultivar. Journal of the science of food and agriculture 1994;64(2):155-61.  https://doi.org/10.1002/jsfa.2740640204
  18. Blois MS. Antioxidant determinations by the use of a stable free radical. Nature 1958;181(4617):1199-200.  https://doi.org/10.1038/1811199a0
  19. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorizationassay. Free Radical Biology and Medicine 1999;26(9-10):1231-7.  https://doi.org/10.1016/S0891-5849(98)00315-3
  20. Park SM, Lee GW, Cho YH. Effect of rheum undulatum extract on antioxidant activity and activity of matrix metalloproteinase-1 in human skin fibroblasts. Journal of Life Science 2008;18(12):1700-4.  https://doi.org/10.5352/JLS.2008.18.12.1700
  21. Schuppan D, Popov Y. Hepatic fibrosis: from bench to bedside. Journal Gastroenterol Hepatol 2002;17(3):300-5.  https://doi.org/10.1046/j.1440-1746.17.s3.18.x
  22. Sanchez-Valle V, Chavez-Tapia NC, Uribe M, Mendez-Sanchez N. Role of oxidative stress and molecular changes in liver fibrosis: a review. Curr Med Chem 2012;19(28):4850-60.  https://doi.org/10.2174/092986712803341520
  23. Parola M. Pinzani M. Liver fibrosis: pathophysiology, pathogenetic targets and clinical issues. Mol Aspect Med 2019;(65):37-55. 
  24. Kren V, Walteriva D. Silybin and Silymarin new effects and applications. Biomed Papers 2005;149(1):29-41.  https://doi.org/10.5507/bp.2005.002
  25. Li Q, Gao J, Pang X, Chen A, Wang Y. Molecular Mechanism of Action of Emodin: As an Anti-Cardiovascular Disease Drug. Frontiers in Pharmacology 2020;11:559607. 
  26. Kim HJ, Kim SY, Shin SP, Yang YJ, Bang CS, Baik GH. Immunological measurement of aspartate/alanine aminotransferase in predicting liver fibrosis and inflammation. Korean J Intern Med 2020;35(2):320-30.  https://doi.org/10.3904/kjim.2018.214
  27. Arriazu E, De Galarreta MR, Cubero FJ, Varela-Rey M, De Obanos MPP, Leung TM, et al. Extracellular Matrix and Liver Disease. Antioxid. Redox Signal 2014;21(7):1078-97.  https://doi.org/10.1089/ars.2013.5697
  28. Bataller R, Brenner DA. Hepatic stellate cells as a target for the treatment of liver fibrosis. Seminars in Liver Disease 2005;25(4):419-31. 
  29. Bataller R, Gines P, Nicolas JM, Gorbig MN, Garcia-Ramallo E, Gasull X, et al. AngiotensinII induces contraction and proliferation of human hepatic stellate cells. Gastroenterology 2000;118(6):1149-56.  https://doi.org/10.1016/S0016-5085(00)70368-4
  30. Bataller R, Schwabe RF, Choi TH, Tang L, Paik YH, Lindquit J, et al. NADPH oxidase signal transduces angiotensin II n in hepatic stellate cells and critical in hepatic fibrosis. J Clin Invest 2003;112(9):1383-94.  https://doi.org/10.1172/JCI18212
  31. De Souza Basso B, Haute GV, Ortega-Ribera M, Luft C, Antunes GL, Bastos MS, et al. Methoxyeugenol deactivates hepatic stellate cells and attenuates liver fibrosis and inflammation through a PPAR-ɣ and NF-kB mechanism. Journal of Ethnopharmacol 2021;280:114433.