Get access

Silybin inhibits interleukin-1β-induced production of pro-inflammatory mediators in canine hepatocyte cultures

Authors

  • A. Y. AU,

    1. Research and Development, Nutramax Laboratories, Inc., Edgewood, MD, USA
    2. Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, NY, USA
    3. Department of Orthopaedic Surgery, Johns Hopkins University, Baltimore, MD, USA
    Search for more papers by this author
  • J. M. HASENWINKEL,

    1. Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, NY, USA
    Search for more papers by this author
  • C. G. FRONDOZA

    1. Research and Development, Nutramax Laboratories, Inc., Edgewood, MD, USA
    2. Department of Orthopaedic Surgery, Johns Hopkins University, Baltimore, MD, USA
    3. College of Veterinary Medicine, Mississippi State University, Mississippi State, MS, USA
    Search for more papers by this author

Carmelita G. Frondoza, PhD, Research and Development, Nutramax Laboratories, Inc., 2208 Lakeside Boulevard, Edgewood, MD 21040, USA. E-mail: cfrondoza@nutramaxlabs.com

Abstract

Au, A. Y., Hasenwinkel, J. M., Frondoza, C. G. Silybin inhibits interleukin-1β-induced production of pro-inflammatory mediators in canine hepatocyte cultures. J. vet. Pharmacol. Therap.34, 120–129.

Hepatocytes are highly susceptible to cytokine stimulation and are fundamental to liver function. We established primary canine hepatocyte cultures to study effects of anti-inflammatory agents with hepatoprotective properties. Hepatocyte cultures were incubated with control media alone, silybin (SB), or the more bioavailable silybin–phosphatidylcholine complex (SPC), followed by activation with interleukin-1 beta (IL-1β; 10 ng/mL). Inflammatory response was measured by prostaglandin E2 (PGE2), interleukin-8 (IL-8), and monocyte chemotactic protein-1 (MCP-1) production and also nuclear factor-kappa B (NF-κB) translocation. Hepatocyte cultures continued production of the phenotypic marker albumin for more than 7 days in culture. IL-1β exposure increased PGE2, IL-8, and MCP-1 production, which was paralleled by NF-κB translocation from the cytoplasm to the nucleus. Pretreatment with SB and SPC significantly inhibited IL-1β-induced production of pro-inflammatory markers and attenuated NF-κB nuclear translocation. We demonstrate for the first time that primary canine hepatocyte cultures can be maintained in culture without phenotypic loss. The observation that hepatocyte cultures respond to pro-inflammatory IL-1β activation indicates hepatocytes as primary cellular targets of extrinsic IL-1β. The ability of SB and SPC to inhibit hepatocyte culture activation by IL-1β reinforces the notion of their hepatoprotective effects. Our primary canine hepatocyte culture model facilitates identification of hepatoprotective agents and their mechanism of action.

Get access to the full text of this article

Ancillary