The Journal of Korean Medicine (대한한의학회지)

The Society of Korean Medicine (대한한의학회)
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Effect of Samhwangsasim-tang and Daehwanghwangryunsasim-tang on Palmitate-induced Lipogenesis in HepG2 cells

Palmitic acid로 지방 축적을 유도한 HepG2 cell에 대한 삼황사심탕과 대황황련사심탕의 효과 연구

  • Um, Eun sik (Department of Clinical Korean Medicine, Graduate School, Kyung Hee University) ;
  • Kim, Young Chul (Department of Clinical Korean Medicine, Graduate School, Kyung Hee University)
  • 엄은식 (경희대학교 대학원 임상한의학과) ;
  • 김영철 (경희대학교 대학원 임상한의학과)
  • Received : 2015.11.27
  • Accepted : 2016.03.24
  • Published : 2016.03.31
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Abstract

Objectives: The goal of this study was to investigate the anti-lipogenic effects of Samhwangsasim-tang(SHT), Daehwanghwangryunsasim-tang(DHT) aqueous extract on HepG2 cells with palmitate. Materials and Methods: HepG2 cells treated with palmitate were used in this study as hepatic steatosis model. Cells were treated with different concentrations of SHT, DHT aqueous extract for 24 hours. Cell viability and cytotoxicity were analyzed by MTT assay. Expressions of Bcl-2, Bax, Survivin, P21, TGF-${\beta}1$, LXR-${\alpha}$, ChREBP, ACC1, SCD1 mRNA were determined by Real-time PCR. Apoptosis of cells was detected by ELISA and FACS. Expression level of caspase-3 was studied by Western blot. Lipid accumulation was indicated by Oil Red O staining. Results: SHT, DHT aqueous extract had no cytotoxicity, but decreased palmitate-induced lipid accumulation in HepG2 cells. SHT aqueous extract suppressed fatty acid synthesis by inhibiting LXR-${\alpha}$, ChREBP, SCD1 activation and increasing TGF-${\beta}1$ expression level. DHT aqueous extract also suppressed fatty acid synthesis by decreasing ChREBP expression and increasing TGF-${\beta}1$ expression. Apoptosis of lipid accumulated cells was increased by enhanced activities of P21, caspase-3 and inhibited expressions of Bcl-2, Survivin. Conclusions: These results suggest that SHT and DHT have an anti-lipogenic effects on lipid accumulation of hepatic cell. Also SHT and DHT have an efficacy to increase apoptosis of adipocyte without cytotoxicity. Therefore, SHT and DHT might have potential clinical applications for treatment of hepatic steatosis.

Keywords

References

  1. Bellentani S, Bedogni G, Miglioli L, Tiribelli C. The epidemiology of fatty liver. European journal of gastroenterology & hepatology, 2004;16(11):1087-1093. https://doi.org/10.1097/00042737-200411000-00002
  2. Marchesini G, Brizi M, Morselli-Labate AM, Bianchi G, Bugianesi E, McCullough AJ, et al. Association of nonalcoholic fatty liver disease with insulin resistance. The American journal of medicine. 1999;107(5):450-455. https://doi.org/10.1016/S0002-9343(99)00271-5
  3. Chalasani N, Younossi Z, Lavine JE. Diehl A M, Brunt EM, Cusi K, et al. The diagnosis and management of non-alcoholic fatty liver disease: Practice Guideline by the American Association for the Study of Liver Diseases, American College of Gastroenterology, and the American Gastroenterological Association. Hepatology. 2012;55(6):2005-2023. https://doi.org/10.1002/hep.25762
  4. Anstee QM, Day CP. The Genetics of Nonalcoholic Fatty Liver Disease: Spotlight on PNPLA3 and TM6SF2. In Seminars in liver disease. 2015;35(3):270-290. https://doi.org/10.1055/s-0035-1562947
  5. Angulo P. GI epidemiology: nonalcoholic fatty liver disease. Alimentary pharmacology & therapeutics. 2007;25(8):883-889. https://doi.org/10.1111/j.1365-2036.2007.03246.x
  6. Ministry of Food and Drug Safety. Influence of dietary intake on non-alcoholic fatty liver disease in Koean. Cheongwon, Korea: Ministry of Food and Drug Safety. 2012
  7. Jang E, Shin MH, Kim KS, Kim Y, Na YC, Woo HJ. Anti-lipoapoptotic effect of Artemisia capillaris extract on free fatty acids-induced HepG2 cells. BMC complementary and alternative medicine. 2014;14(1):253. https://doi.org/10.1186/1472-6882-14-253
  8. Browning JD, Horton JD. Molecular mediators of hepatic steatosis and liver injury. Journal of Clinical Investigation. 2004;114(2):147. https://doi.org/10.1172/JCI200422422
  9. Park SH. Nonalcoholic fatty liver disease: treatment. Korean Journal of Medicine. 2010;79(5):481-489.
  10. Han CW, Lee JH. Effects of KHchunggan-tang on the nonalcoholic fatty liver disease in palmitate-induced cellular model. J Korean Oriental Med. 2011;32(1):109-110.
  11. Kim SY, Kwon JN, Lee I, Hong JW, Choi JY, Park SH, et al. Research on Anti-lipogenic Effect and Underlying Mechanism of Laminaria japonica on Experimental Cellular Model of Non-alcoholic Fatty Liver Disease. The Journal of Korean Oriental Internal Medicine. 2014;35(2):175-183.
  12. Choi MY, Woo HJ, Kim YC, Lee JH. Effects of Gamisaenggan-tang on High Fat Diet-induced Nonalcoholic Fatty Liver Disease. Korean J. Orient. Int. Med. 2009;30(2):365-374.
  13. Lee JY, Yoon BK. Effects of Samhwangsasim-tang on obesity-related metabolic diseaseinmice. Dept. of Korean Medicine. 2014;22(1):93-104.
  14. Yoon HJ, Park YS. Effects of Scutellariabaicalensis water extract on lipid metabolism and antioxidant defense system in rats fed high fat diet. Journal of The Korean Society of Food Science and Nutrition. 2010.
  15. Ro HS, Ko WK, Kim OJ, Park KK, Cho YW, Park HS. Antihyperlipidemic Activity of Scutellarisbaicalensis Georg., Coptidis japonica Makino and RheikoreanumNakai on Experimental Hyperlipidemia in Rats. Journal of Pharmaceutical Investigation. 1996;26(3):215-219.
  16. Choi SE, Jung IR, Lee YJ, Lee SJ, Lee JH, Kim Y, et al. Stimulation of lipogenesis as well as fatty acid oxidation protects against palmitate-induced INS-1 ${\beta}$-cell death. Endocrinology. 2011;152(3):816-827. https://doi.org/10.1210/en.2010-0924
  17. Ramirez-Zacarias JL, Castro-Munozledo F, Kuri-Harcuch W. Quantitation of adipose conversion and triglycerides by staining intracytoplasmic lipids with Oil Red O. Histochemistry. 1992;97(6):493-497. https://doi.org/10.1007/BF00316069
  18. Samuel VT, Liu ZX, Qu X, Elder BD, Bilz S, Befroy D, et al. Mechanism of hepatic insulin resistance in non-alcoholic fatty liver disease. Journal of Biological Chemistry. 2004;279(31):32345-32353. https://doi.org/10.1074/jbc.M313478200
  19. Park SH, Jeon WK, Kim SH, Kim HJ, Park DI, Cho YK. Prevalence and risk factors of nonalcoholic fatty liver disease among Korean adults. Journal of gastroenterology and hepatology. 2006;21(1):138-143. https://doi.org/10.1111/j.1440-1746.2005.04086.x
  20. Marra F, Gastaldelli A, Baroni GS, Tell G, Tiribelli C. Molecular basis and mechanisms of progression of non-alcoholic steatohepatitis. Trends in molecular medicine. 2008;14(2):72-81. https://doi.org/10.1016/j.molmed.2007.12.003
  21. Wahren J, Sato Y, Ostman J, Hagenfeldt L, Felig P. Turnover and splanchnic metabolism of free fatty acids and ketones in insulin-dependent diabetics at rest and in response to exercise. Journal of Clinical Investigation. 1984;73(5):1367. https://doi.org/10.1172/JCI111340
  22. Chen G, Liang G, Ou J, Goldstein JL, Brown MS. Central role for liver X receptor in insulin-mediated activation of Srebp-1c transcription and stimulation of fatty acid synthesis in liver. Proceedings of the National Academy of Sciences of the United States of America. 2004;101(31):11245-11250. https://doi.org/10.1073/pnas.0404297101
  23. Chung MH, Han SJ. Effect of composite preparation of crude drugs on experimentally induced hyperlipemia in rats-Sam WhangSasim Tang and Whang Ryun Haedok Tang. Korean Journal of Pharmacognosy (Korea Republic). 1996;7(2):46-54
  24. Oh MG, Song YS. Influence of RhizomaRhei water extract on the obese mouse model. J Oriental Rehab Med. 1997;7(2):46-54.
  25. Oltval ZN, Milliman CL, Korsmeyer SJ. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programed cell death.cell. 1993;74(4):609-619. https://doi.org/10.1016/0092-8674(93)90509-O
  26. Tamm I, Wang Y, Sausville ED, Scudiero DA, Vigna N, Oltersdorf T, et al. IAP-family protein survivin inhibits caspase activity and apoptosis induced by Fas (CD95), Bax, caspases, and anticancer drugs.Cancer research. 1998;58(23):5315-5320.
  27. Levkau B, Koyama H, Raines EW, Clurman B E, Herren B, Orth K, et al. Cleavage of p21 Cip1/Waf1 and p27 Kip1 mediates apoptosis in endothelial cells through activation of Cdk2: role of a caspase cascade. Molecular cell. 1998;1(4):553-563. https://doi.org/10.1016/S1097-2765(00)80055-6
  28. Porter AG, Janicke RU. Emerging roles of caspase-3 in apoptosis. Cell death and differentiation. 1996;6(2):99-104. https://doi.org/10.1038/sj/cdd/4400476
  29. Petruschke TH, Rohrig K, Hauner H. Transforming growth factor beta (TGF-beta) inhibits the differentiation of human adipocyte precursor cells in primary culture. International journal of obesity and related metabolic disorders: journal of the International Association for the Study of Obesity. 1994;18(8):532-536.
  30. Nam IK, Yoo J. Downregulation of SGK1 Expression is Critical for TGF-${\beta}$-induced Apoptosis in Mouse Hepatocytes Cells. Journal of life science. 2012;22(11):1500-1506. https://doi.org/10.5352/JLS.2012.22.11.1500
  31. Cha JY, Repa JJ. The liver X receptor (LXR) and hepatic lipogenesis The carbohydrate-response element-binding protein is a target gene of LXR. Journal of Biological Chemistry. 2007;282(1):743-751. https://doi.org/10.1074/jbc.M605023200
  32. Berlanga A, Guiu-Jurado E, Porras JA, Auguet T. Molecular pathways in non-alcoholic fatty liver disease. Clinical and experimental gastroenterology. 2014;7:221.
  33. Repa JJ, Liang G, Ou J, Bashmakov Y, Lobaccaro JMA, Shimomura I. Regulation of mouse sterol regulatory element-binding protein-1c gene (SREBP-1c) by oxysterol receptors, LXR${\alpha}$ and LXR${\beta}$. Genes & development. 2000;14(22):2819-2830. https://doi.org/10.1101/gad.844900