Anti-Serum Amyloid A Anticorps [607] (A32856)

OBJECTIVE:

Obesity is associated with monocyte-macrophage accumulation in adipose tissue. Previously, we showed that glucose-stimulated production by adipocytes of serum amyloid A (SAA), monocyte chemoattractant protein (MCP)-1, and hyaluronan (HA) facilitated monocyte accumulation. The current objective was to determine how the other major nutrient, free fatty acids (FFAs), affects these molecules and monocyte recruitment by adipocytes.

RESEARCH DESIGN AND METHODS:

Differentiated 3T3-L1, Simpson-Golabi-Behmel syndrome adipocytes, and mouse embryonic fibroblasts were exposed to various FFAs (250 micromol/l) in either 5 or 25 mmol/l (high) glucose for evaluation of SAA, MCP-1, and HA regulation in vitro.

RESULTS:

Saturated fatty acids (SFAs) such as laurate, myristate, and palmitate increased cellular triglyceride accumulation, SAA, and MCP-1 expression; generated reactive oxygen species (ROS); and increased nuclear factor (NF) kappaB translocation in both 5 and 25 mmol/l glucose. Conversely, polyunsaturated fatty acids (PUFAs) such as arachidonate, eicosapentaenate, and docosahexaenate (DHA) decreased these events. Gene expression could be dissociated from triglyceride accumulation. Although excess glucose increased HA content, SFAs, oleate, and linoleate did not. Antioxidant treatment repressed glucose- and palmitate-stimulated ROS generation and NFkappaB translocation and decreased SAA and MCP-1 expression and monocyte chemotaxis. Silencing toll-like receptor-4 (TLR4) markedly reduced SAA and MCP-1 expression in response to palmitate but not glucose. DHA suppressed NFkappaB translocation stimulated by both excess glucose and palmitate via a peroxisome prolifterator-activated receptor (PPAR) gamma-dependent pathway.

CONCLUSIONS:

Excess glucose and SFAs regulate chemotactic factor expression by a mechanism that involves ROS generation, NFkappaB, and PPARgamma, and which is repressed by PUFAs. Certain SFAs, but not excess glucose, trigger chemotactic factor expression via a TLR4-dependent pathway.

INTRODUCTION:

Serum amyloid A (SAA) and C-reactive protein (CRP) are both acute-phase reactants synthesized by the liver upon stimulation by proinflammatory cytokines reflecting both the acute and chronic inflammatory states.

METHODS:

We have established a one-step, sandwich ELISA on microplate for SAA using commercial antibodies for coating and detection.

RESULTS:

This in-house ELISA has a sensitivity of 0.12 mg/l. Both within-day and between-day CVs were <10%. The in-house assay correlated well with the commercial ELISA kit from Anogen (r=0.95). We also established the reference range for apparently healthy Chinese. Statistically higher SAA values were found in those >50 years old. No difference was found between genders. We found only slightly increased levels of SAA in early stage of type 2 diabetics, but highly increased levels of SAA were detected in patients with acute myocardial infarction, generally associated with intense inflammation. At the early stage of type 2 diabetes associated with low inflammation, SAA was found to be complementary to CRP in test sensitivity.

CONCLUSIONS:

Based on our data and reports from the literature we believe that SAA responds differently than CRP in inflammatory diseases such as in type 2 diabetes and acute myocardial infarction, and is complementary to CRP in test sensitivity.