In vivo characterization of inflammatory biomarkers in swine and the impact of flunixin meglumine administration.
Non-steroidal anti-inflammatory drugs (NSAID) are a family of chemicals that function to reduce pain, fever, and inflammation, and they are commonly used in people and animals for this purpose. Currently there are no NSAIDs approved for the management of inflammation in swine due to a lack of validated animal models and suitable biomarkers to assess efficacy. A previous in vitro study examining biomarkers of inflammation identified fourteen genes that were significantly altered in response to Escherichia coli lipopolysaccharide (LPS)-induced inflammation. In the present study, five of those fourteen genes were tested in vivo to determine if the same effects observed in vitro were also observed in vivo. Plasma levels of prostaglandin E(2) (PGE(2)), an essential mediator of fever and inflammation, were also determined. Two groups of swine were stimulated with LPS with the second group also treated with flunixin meglumine. Blood was collected at 0, 1, 3, 6, 8, 24, and 48 h post LPS-stimulation. The RNA was extracted from the blood and quantitative real-time-PCR (qRT-PCR) was utilized to determine the expression patterns of CD1, CD4, serum amyloid A2 (SAA2), Caspase 1, and monocyte chemoattractant protein 1 (MCP-1). The LPS-stimulated animals demonstrated a statistically significant alteration in expression of SAA2 and CD1 at 3h post-stimulation. Flunixin meglumine treated animals' demonstrated reduced expression of CD1 in comparison to the LPS-stimulated swine at 24 and 48 h post LPS-stimulation. Flunixin meglumine treated animals exhibited reduced expression of SAA2 at 48 h post-stimulation compared to LPS-stimulated swine. Swine treated with LPS demonstrated statistically significant increases in plasma PGE(2) at 1h post-stimulation. Swine treated with flunixin meglumine had no increase in plasma PGE(2) levels at any time. These results demonstrate that PGE(2) production, along with two out of five genes (SAA2 and CD1) have the potential to serve as early biomarkers of inflammation as well as indicators of NSAID efficacy.
U.S. FDA, Center for Veterinary Medicine, Office of Research, Laurel, MD 20708, USA.
Sharla M
SM
Peters
Haile
H
Yancy
Christine
C
Deaver
Yolanda L
YL
Jones
Elizabeth
E
Kenyon
Oscar A
OA
Chiesa
Juan
J
Esparza
Rudell
R
Screven
Vicki
V
Lancaster
John T
JT
Stubbs
Maocheng
M
Yang
Paddy L
PL
Wiesenfeld
Michael J
MJ
Myers
D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D018949:Antigens, CD1 / Q000097:blood; D015415:Biomarkers / Q000097:blood; D015704:CD4 Antigens / Q000097:blood; D020170:Caspase 1 / Q000097:blood; D018932:Chemokine CCL2 / Q000097:blood; D003002:Clonixin / Q000031:analogs & derivatives* / Q000494:pharmacology; D015232:Dinoprostone / Q000097:blood; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D007249:Inflammation / Q000097:blood / Q000188:drug therapy / Q000662:veterinary*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D000685:Serum Amyloid A Protein / Q000032:analysis; D013552:Swine; D013553:Swine Diseases / Q000097:blood* / Q000188:drug therapy / Q000276:immunology; D013929:Thromboxane B2 / Q000097:blood
D000894:Anti-Inflammatory Agents, Non-Steroidal; D018949:Antigens, CD1; D015415:Biomarkers; D015704:CD4 Antigens; D018932:Chemokine CCL2; D008070:Lipopolysaccharides; D000685:Serum Amyloid A Protein; D013929:Thromboxane B2; C014558:flunixin meglumine; D020170:Caspase 1; D015232:Dinoprostone; D003002:Clonixin