Anti-inflammatory effects of osthole in peripheral blood mononuclear cells from Hanwoo (Bos taurus coreanae)

  • Kim, Seung-Chang (Animal Genomics & Bioinformatics Division, National Institute of Animal Science, Rural Development Administration) ;
  • Lee, Seung-Hwan (Division of Animal and Dairy Science, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Chai, Han-Ha (Animal Genomics & Bioinformatics Division, National Institute of Animal Science, Rural Development Administration) ;
  • Kim, Ui-Hyung (Hanwoo Research Institute, National Institute of Animal Science, Rural Development Administration) ;
  • Chung, Ki-Yong (Hanwoo Research Institute, National Institute of Animal Science, Rural Development Administration) ;
  • Jang, Sun-Sik (Hanwoo Research Institute, National Institute of Animal Science, Rural Development Administration) ;
  • Choi, Bong-Hwan (Animal Genomics & Bioinformatics Division, National Institute of Animal Science, Rural Development Administration)
  • Received : 2019.03.25
  • Accepted : 2019.07.16
  • Published : 2019.09.01


Due to the ban on the use of antibiotics, interest has been increasing for the development of therapeutic agents to treat various diseases using natural resources. Osthole, a natural coumarin compound used in traditional Chinese medicines, exerts an anti-inflammatory effect, but its effects in cows remain unknown. In this study, the effect of osthole on lipopolysaccharide (LPS)- or concanavalin-A (Con-A)- stimulated peripheral blood mononuclear cells (PBMCs) was assessed. Jugular venous blood was collected from Korean calves, and PBMCs were isolated. They were then used to study the immune response of PBMCs to treatment with osthole and LPS or Con-A for 72 h by measuring inflammatory cytokines including tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$) and interferon-${\gamma}$ ($IFN-{\gamma}$). Osthole significantly inhibited the mRNA secretion of $TNF-{\alpha}$ and $IFN-{\gamma}$ in a dose-dependent manner. Therefore, osthole inhibited LPS- or Con-A- induced $TNF-{\alpha}$ and Con-A-induced $IFN-{\gamma}$ production significantly in dose-dependent manner. These results clearly suggest that osthole inhibited the LPS- or Con-A- stimulated upregulation of pro-inflammatory cytokines in a dose-dependent manner, without causing obvious cytotoxic effects. Osthole could also protect cows from LPS- or Con-A- induced endotoxin shock, possibly by inhibiting the production of pro-inflammatory cytokines, which suggests that osthole might be a novel therapeutic agent for the prevention of inflammatory diseases.


Grant : Genetic variation and gene expression in a parental origin-specific manner for Selection of Hanwoo cows

Supported by : Rural Development Administration


  1. Abbas KA, Lichtman HA. 2012. Basic immunology. Elsevier, Amsterdam, Netherlands.
  2. Adams JL, Czuprynski CJ. 1990. Bacterial lipopolysaccharide induces release of tumor necrosis factoralpha from bovine peripheral blood monocytes and alveolar macrophages in vitro. Journal of Leukocyte Biology 48:549-556.
  3. Bernard GR, Artigas A, Brigham KL, Carlet J, Falke K, Hudson L, Lamy M, Legall JR, Morris A, Spragg R. 1994. The American-European consensus conference on ARDS. Definitions, mechanisms, relevant outcomes, and clinical trial coordination. American Journal of Respiratory and Critical Care Medicine 149:818-824. doi:10.1164/ajrccm.149.3.7509706
  4. Chiu PR, Lee WT, Chu YT, Lee MS, Jong YJ, Hung CH. 2008. Effect of the Chinese herb extract osthol on IL-4-induced eotaxin expression in BEAS-2B cells. Pediatrics and Neonatology 49:135-140. doi:10.1016/S1875-9572(08)60028-5
  5. Dinarello CA. 1996. Cytokines as mediators in the pathogenesis of septic shock. Current Topics in Microbiology and Immunology 216:133-165.
  6. Gantner F, Leist M, Lohse AW, Germann PG, Tiegs G. 1995. Concanavalin A-induced T-cell-mediated hepatic injury in mice: The role of tumor necrosis factor. Hepatology 21:190-198.
  7. Gunther J, Esch K, Poschadel N, Petzl W, Zerbe H, Mitterhuemer S, Blum H, Seyfert HM. 2011. Comparative kinetics of Escherichia coli- and Staphylococcus aureus-specific activation of key immune pathways in mammary epithelial cells demonstrates that S. aureus elicits a delayed response dominated by interleukin-6 (IL-6) but not by IL-1A or tumor necrosis factor alpha. Infection and Immunity 79:695-707. doi:10.1128/IAI.01071-10
  8. Guzik TJ, Korbut R, Adamek-Guzik T. 2003. Nitric oxide and superoxide in inflammation and immune regulation. Journal of Physiology and Pharmacology 54:469-487.
  9. Hoult JR, Paya M. 1996. Pharmacological and biochemical actions of simple coumarins: Natural products with therapeutic potential. General Pharmacology 27:713-722.
  10. Jung WK, Lee DY, Choi YH, Yea SS, Choi I, Park SG, Seo SK, Lee SW, Lee CM, Kim SK, Jeon YJ, Choi IW. 2008. Caffeic acid phenethyl ester attenuates allergic airway inflammation and hyperresponsiveness in murine model of ovalbumin-induced asthma. Life Sciences 82:797-805. doi:10.1016/j.lfs.2008.01.014
  11. Li F, Gong Q, Wang L, Shi J. 2012. Osthole attenuates focal inflammatory reaction following permanent middle cerebral artery occlusion in rats. Biological & Pharmaceutical Bulletin 35:1686-1690.
  12. Liao PC, Chien SC, Ho CL, Wang EI, Lee SC, Kuo YH, Jeyashoke N, Chen J, Dong WC, Chao LK, Hua KF. 2010. Osthole regulates inflammatory mediator expression through modulating NF-kappaB, mitogenactivated protein kinases, protein kinase C, and reactive oxygen species. Journal of Agricultural and Food Chemistry 58:10445-10451. doi:10.1021/jf102812t
  13. Livak KJ, Schmittgen TD. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-delta delta C(T)) method. Methods 25:402-408. doi:10.1006/meth.2001.1262
  14. Luheshi G, Rothwell N. 1996. Cytokines and fever. International Archives of Allergy and Immunology 109:301-307. doi:10.1159/000237256
  15. Nagai Y, Nochi T, Watanabe K, Watanabe K, Aso H, Kitazawa H, Matsuzaki M, Ohwada S, Yamaguchi T. 2005. Localization of interleukin-18 and its receptor in somatotrophs of the bovine anterior pituitary gland. Cell and Tissue Research 322:455-462. doi:10.1007/s00441-005-0016-0
  16. Nakamura T, Kodama N, Arai Y, Kumamoto T, Higuchi Y, Chaichantipyuth C, Ishikawa T, Ueno K, Yano S. 2009. Inhibitory effect of oxycoumarins isolated from the Thai medicinal plant Clausena guillauminii on the inflammation mediators, iNOS, TNF-alpha, and COX-2 expression in mouse macrophage RAW 264.7. Journal of Natural Medicines 63:21-27. doi:10.1007/s11418-008-0277-5
  17. Okamoto T, Kawasaki T, Hino O. 2003. Osthole prevents anti-Fas antibody-induced hepatitis in mice by affecting the caspase-3-mediated apoptotic pathway. Biochemistry & Pharmacology 65:677-681.
  18. Okamoto T, Kobayashi T, Yoshida S. 2007. Synthetic derivatives of osthole for the prevention of hepatitis. Medicinal Chemistry 3:35-44.
  19. Panadero R, Dacal V, Lopez C, Vazquez L, Cienfuegos S, Diaz P, Morrondo P, Diez-Banos P. 2009. Immunomodulatory effect of Hypoderma lineatum antigens: In vitro effect on bovine lymphocyte proliferation and cytokine production. Parasite Immunology 31:72-77. doi:10.1111/j.1365-3024.2008.01072.x
  20. Son JY, Renchinkhan G, Bae HC, Paik SH, Lee JY, Nam MS. 2018. Cytokine modulation in Raw 264.7 macrophages treated with ginseng fermented by Penibacillus MBT213. Korean Journal of Agricultural Science 45:769-777. [in Korean]
  21. Tang CH, Yang RS, Chien MY, Chen CC, Fu WM. 2008. Enhancement of bone morphogenetic protein-2 expression and bone formation by coumarin derivatives via p38 and ERK-dependent pathway in osteoblasts. European Journal of Pharmacology 579:40-49. doi:10.1016/j.ejphar.2007.10.013
  22. Teng CM, Lin CH, Ko FN, Wu TS, Huang TF. 1994. The relaxant action of osthole isolated from Angelica pubescens in guinea-pig trachea. Naunyn-Schmiedeberg's Archives of Pharmacology 349:202-208.
  23. Utsunomiya I, Ito M, Oh-ishi S. 1998. Generation of inflammatory cytokines in zymosan-induced pleurisy in rats: TNF induces IL-6 and cytokine-induced neutrophil chemoattractant (CINC) in vivo. Cytokine 10:956-963. doi:10.1006/cyto.1998.0376
  24. Van Amersfoort ES, Van Berkel TJ, Kuiper J. 2003. Receptors, mediators, and mechanisms involved in bacterial sepsis and septic shock. Clinical Microbiology Reviews 16:379-414.
  25. Walker PD. 1992. Bacterial vaccines: Old and new, veterinary and medical. Vaccine 10:977-990.
  26. Yang LL, Wang MC, Chen LG, Wang CC. 2003. Cytotoxic activity of coumarins from the fruits of Cnidium monnieri on leukemia cell lines. Planta Medica 69:1091-1095. doi:10.1055/s-2003-45188
  27. You L, Feng S, An R, Wang X. 2009. Osthole: A promising lead compound for drug discovery from a traditional Chinese medicine (TCM). Natural Product Communications 4:297-302.
  28. Zhang X, Wei HX, Rui S, Wei H, Tian Z. 2010. Opposite effects of high and low doses of interleukin-2 on T cell-mediated hepatitis in mice (interleukin-2 on hepatitis). Hepatology International 4:641-648. doi:10.1007/s12072-010-9196-0