DOI QR코드

DOI QR Code

Hepatoprotective Effects of Sumaeyaksuk (Artemisia argyi H.) Extract on LPS-mediated Inflammatory Response

LPS에 의해 유도된 염증반응에서 섬애약쑥 추출물의 간보호 효과

  • Received : 2016.06.27
  • Accepted : 2016.08.08
  • Published : 2016.11.30

Abstract

Artemisia, a plant widely used as traditional herbal medicine in many countries, has drawn attention of the researchers. And its extracts or compounds are known to have an efficacy of antioxidant, anti-diabete, anti-cancer, anti-inflammation and neuroprotection. Sumaeyaksuk is a variant of the Artemisia argyi and major constituents are eupatilin and jaceosidin. This study was performed to investigate the effects of the sumaeyaksuk aqueous extract on inflammatory response induced by lipopolysaccharide (LPS) in human hepatoma HepG2 cells. To examine the potential hepatoprotective properties of sumaeyaksuk extract, cell viability, as well as nitric oxide (NO), reactive oxygen species (ROS), macrophage colony-stimulating factor (M-CSF), interleukin-8 (IL-8) levels, alanine transaminase (ALT), and aspartate transaminase (AST) activities, were measured. Cytotoxic activity of extracts on HepG2 cells was measured by MTT assay. Sumaeyaksuk extract did not induce cytotoxicity at concentrations of $0{\sim}400{\mu}g/mL$. NO and ROS levels significantly decreased with increasing concentration of the extract. The secretion levels of M-CSF and IL-8 were suppressed by sumaeyaksuk extract in a dose-dependent manner. Moreover, ALT (75.4%) and AST (61.6%) levels significantly decreased in sumaeyaksuk extract-treated cells at $400{\mu}g/mL$. These results suggested that the sumaeyaksuk extract attenuates the LPS-induced hepatotoxicity resulting from regulation of inflammatory factors and could potentially be used as a hepatitis therapeutic agent.

본 연구는 섬애약쑥의 생리활성 규명을 위한 연구의 일환으로 물 추출물의 항염증 활성 및 간 염증에 대한 보호효과를 세포수준에서 확인하였다. 세포독성이 없는 $100{\sim}400{\mu}g/ml$의 농도 범위에서 섬애약쑥 추출물은 HepG2 세포에서 nitric oxide (NO)와 reactive nitrogen species (ROS)생성을 농도의존적으로 저해하였으며, 간염질환과 밀접한 관계가 있는 cytokine인 M-CSF와 IL-8의 발현을 감소시켰다. 또한 직접적인 간 손상을 나타낼 수 있는 지표인 AST와 ALT의 발현을 유의성 있게 감소시키는 결과를 얻을 수 있었다. 본 실험을 통해서 섬애약쑥물 추출물은 세포독성이 없는 $200{\sim}400{\mu}g/ml$의 범위에서 농도의존적으로 간 염증의 개선 효과를 가질 것이라는 단서를 확인하였다.

Keywords

References

  1. Akashi, S., Nagai, Y., Ogata, H., Oikawa, M., Fukase, K. and Kusumoto, S. 2001. Human MD-2 confers on mouse Toll-like receptor 4 species-specific lipopolysaccharide recognition. Int. Immunol. 13, 1595-1599. https://doi.org/10.1093/intimm/13.12.1595
  2. Billiar, T. R., Curran, R. D., Stuehr, D. J., Ferrari, F. K. and Simmons, R. L. 1989. Evidence that the activation of kupffer cells results in the production of L-arginine metabolites thar release cell-associated iron and inhibit hepatocyte proein synthesis. Surgery 106, 364-368.
  3. Choi, M. W. and Kim, J. I. 2013. Anti-inflammatory effect of ethyl acetate fraction isolated from undaria pinnatifida on lipopolysaccharides-stimulated RAW 264.7 Cells. Kor. J. Fish. Aquat. Sci. 46, 384-392.
  4. Chung, K. F. 2006. Cytokines as targets in chronic obstructive pulmonary disease. Curr. Drug Targets 7, 675-681. https://doi.org/10.2174/138945006777435263
  5. Elewa, H., Abd-Elmeneem, M., Hashem, A. M. and Alshehaby, A. 2010. Study of interleukin-8 (IL-8) serum level in patients with chronic liver disease due to hepatitis C virus (HCV) with and without hepato cellular carcinoma (HCC). Int. J. Hepatol. 1, 9-17.
  6. Fonseca, S. G., Romao, P. R., Figueiredo, F., Morais, R. H., Lima, H. C., Ferreira, S. H. and Cunha, F. Q. 2003. TNF-alpha mediates the induction of nitric oxide synthase in macrophages but not in neutrophils in experimental cutaneous leishmaniasis. Eur. J. Immunol. 33, 2297-2306. https://doi.org/10.1002/eji.200320335
  7. Gabriel, L. P. and William, R. H. and Hayes, A. W. 1982. Principles and Methods of Toxicology. Raben. Press. 407-445.
  8. Giustarini, D., Rossi, R., Milzani, A. and Dalle-Donne, I. 2008. Nitrite and nitrate measurement by Griess reagent in human plasma: evaluation of interferences and standardization. Methods Enzymol. 440, 361-380.
  9. Halliwell, B. 1990. How to characterize a biological antioxidant. Free. Radic. Res. Commun. 9, 1-13. https://doi.org/10.3109/10715769009148569
  10. Han, E. K., Jin, Y. X., Yoo, Y. S., Jung, E. J., Lee, J. Y. and Chung, C. K. 2009. Effect of Artemisia capillaris and Paecilomyces japonicaon the reduction of hepatotoxicity and lipid metabolism induced by ethanol. J. Kor. Soc. Food Sci. Nutr. 38, 1016-1023. https://doi.org/10.3746/jkfn.2009.38.8.1016
  11. Haw, I. W., Lee, S. D. and Hwang, W. I. 1985. A study on the nutritional effects in rats by feeding basal diet supplemented with mugwort powder. J. Kor. Soc. Food Nutr. 14, 123-130.
  12. Huang, Y. S., Hwang, S. J., Chan, C. Y., Wu, J. C., Chao, Y., Chang, F. Y. and Lee, S. D. 1999. Serum levels of cytokines in hepatitis Crelate dliver disease: a longitudinal study. Zhonghua. Yi XueZa. Zhi. 62, 327-333.
  13. Itoh, Y., Okanoue, T., Enjyo, F., Sakamoto, S., Ohmoto, Y., Hirai, Y., Kagawa, K. and Kashima, K. 1994. Serum levels of macrophage colony stimulating factor (M-CSF) in liver disease. J. Hepatol. 21, 527-535. https://doi.org/10.1016/S0168-8278(94)80097-9
  14. Joung, H. S. 1993. A study on the sensory of Ssooksulgis added with mugworts. J. East Asian Soc. Dietary Life 2, 175-180.
  15. Jung, D. Y. and Park, S. W. 2002. Cytotoxicity of water fraction of Artermisia argyi against L1210 cells and antioxidant enzyme activities. J. Pharm. Soc. Korea 46, 39-46.
  16. Kim, O. K. 2001. Protective effects of extracts of Diospyrus kaki folium against hepatotoxicity in carbon tetrachloride intoxicated rats. J. Kor. Soc. Food Sci. Nutr. 30, 97-101.
  17. Kim, M. H., Lee, S. D. and Ryu, C. K. 1985. A study on the nutritional effects of boiling water extracts of mugwort powder in rats. J. Kor. Soc. Food Nutr. 14, 131-136.
  18. Lee, G. D., Kim, J. S, Bae, J. O. and Yoon, H. S. 1992. Antioxidative effectiveness of water extract and ether extract in worm-wood (Artemisia montana Pampan). J. Kor. Soc. Food Nutr. 21, 17-22.
  19. Lee, H. J. 2010. Evaluation of the quality characteristics of sponge cake containing mugwort powder. J. East Asian Soc. Dietary Life. 20, 95-102.
  20. Lee, Y. H., Ho, J. N., Dong, M. S., Park, J. H., Kim, H. K., Hong, B. S., Shin, D. H. and Cho, H. Y. 2005. Transfected HepG2 cells for evaluation of catechin effects on alcohol-induced CYP2E1 cytotoxicity. J. Microbiol. Biotechnol. 15, 1310-1316.
  21. Lieber, C. S. 1995. Medical disorders of alcoholism. N. Engl. J. Med. 333, 1058-1065. https://doi.org/10.1056/NEJM199510193331607
  22. Lieber, C. S. 2004. The unexpected outcomes of medical research: serendipity and the microsomal ethanol oxidizing system. J. Hepatol. 40, 198-202. https://doi.org/10.1016/j.jhep.2003.12.005
  23. McPhalen, C. A., Vincent, M. G. and Jansonius, J. N. 1992. X-ray structure refinement and comparison of three forms of mitochondrial aspartate aminotransferase. J. Mol. Biol. 225, 495-517. https://doi.org/10.1016/0022-2836(92)90935-D
  24. Mitchell, J. A., Kohlhaas, K. L. and Sorrentino, R. 1993. Induction by endoxin of nitric oxide synthase in the rat mesentery: lack of effect on action of vasoconstictors. Br. J. Pharmacol. 109, 265-270. https://doi.org/10.1111/j.1476-5381.1993.tb13563.x
  25. Morrison, D. C. and Ulevitch, R. J. 1978. The effects of bacterial endotoxins on host mediation systems. Am. J. Pathol. 93, 526-617.
  26. Park, W. S. and Kim, D. H. 2008. Effect of mixture of fermented Artemisiae Argi Folium and fermented sophorae radix on hydrogen peroxide production within mouse macrophage raw 264.7 with EtOH and nicotine. Kor. J. Ori. Physiol. Pathol. 22, 1293-1298.
  27. Rhee, K. J., Kim, H. C. and Jung, B. D. 2012. Effect of Ginkgo biloba extract on the survival rate in lipopolysaccharide-induced sepsis model mice. Kor. J. Vet. Serv. 35, 191-195. https://doi.org/10.7853/kjvs.2012.35.3.191
  28. Russwurm, S., Stonans, I., Stonane, E., Weigand, G., Wiederhold, M., Jager, L. and Reinhart, K. 1998. HepG2 hepatocytes express IFN-${\gamma}$, TNF-${\alpha}$, TGF-${\beta}$, M-CSF, oncostatin-M, ICAM-1, IL-4, IL-5, IL-7, IL-10, IL-11, IL-12 and IL-6 receptor genes in vitro. Crit. Care. 2, 5-15. https://doi.org/10.1186/cc135
  29. Ryakhovskaya, T. V., Ushbaeva, G. G. and Zhemaletdinov, F. G. 1989. The antitumor activity of phenol compounds from some Artemisia spp. L. Rastit. Resur. 25, 249-253.
  30. Shim, Y. J., Paik, J. E. and Chun, H. J. 1991. A study on the texture characteristics of Ssooksulgis affected by mugworts. Kor. J. Soc. Food Sci. 7, 35-43.
  31. Shimamura, M. and Mieda, Y. 1993. Changes in free and bound alcohol metabolitesin the urine during ethanol oxidation. Arukoru. Kenkyuto. Yakubutsu Ison. 28, 441-452.
  32. Sim, Y. J., Han, Y. S. and Chun, H. J. 1992. Studies on the nutritional components of mugwort, Artemisia mongolica Fischer. Kor. J. Food Sci. Technol. 24, 49-53.
  33. Stonans, I., Stonane, E., Russwurm, S., Deigner, H. P., Bohm, K. J., Wiederhold, M., Jager, L. and Reinhart, K. 1999. HepG2 human hepatoma cells express multiple cytokine genes. Cytokine 11, 151-156. https://doi.org/10.1006/cyto.1998.0366
  34. Sugawara, S., Arakaki, R., Rikiishi, H. and Takada, H. 1999. Lipoteichoic acid acts as an antagonist and an agonist of lipopolysaccharide on human gingival fibroblasts and monocytes in a CD14-dependent manner. Infect. Immun. 67, 1623-16 32.
  35. Takeda, K., Kaisho, T. and Akira, S. 2003. Toll-like receptors. Annu. Rev. Immunol. 21, 335-376. https://doi.org/10.1146/annurev.immunol.21.120601.141126
  36. Tsukamoto, S., Kanegae, T., Uchigasaki, S., Kitazawa, M., Fujioka, T., Fujioka, S., Imamura, Y. and Nagoya, T., 1993. Changes in free and bound alcohol metabolitesin the urine during ethanol oxidation. Arukoru. Kenkyuto. Yakubutsu Ison. 28, 441-452.
  37. Uehara, A., Sugawara, S., Tamai, R. and Takada, H. 2001. Contrasting responses of human gingival and colonic epithelial cells to lipopolysaccharides, lipoteichoic acids and peptidoglycans in the presence of soluble CD14. Med. Microbiol. Immunol (Berl). 189, 185-192. https://doi.org/10.1007/s004300100063
  38. Xaus, J. M., Comalada, A. F., Valledor, J., Lloberas, F., Lopez-Soriano, J. M., Argiles, C. and Celada, A. 2000. LPS induces apoptosis in macrophages mostly through the autocrine production of TNF-${\alpha}$. Blood 95, 3823-3831.
  39. Zimmermann, H. W., Seidler, S., Gassler, N., Nattermann, J., Luedde, T., Trautwein, C. and Tacke, F. 2011. Interleukin-8 is activated in patients with chronic liver diseases and associated with hepatic macrophage accumulation in human liver fibrosis. PLoS One 6, e21381. https://doi.org/10.1371/journal.pone.0021381

Cited by

  1. Antioxidant and anti-inflammatory activities of Opuntia ficus-indica and Opuntia humifusa fruits ethanol extracts vol.25, pp.5, 2018, https://doi.org/10.11002/kjfp.2018.25.5.586