The Effects Of Acylation Stimulating Protein On The Inflammatory In 3T3-L1 Adipocytes

ObjectiveTo study the changes of the inflammatory pathway in 3T3-L1 adipocytes ,under the ASP stimulation after induce insulin resistance by free fatty acids(FFA) .Then to explore whether ASP work on the inflammatory pathway and the mechanism of it .Methods1.Classic hormone cocktail(10μg/ml insulin, 0.5 mmol/L 1 - methyl-3 - isobutyl xanthine and 1.0μmol/L dexamethasone) induction method to induce 3T3-L1 preadipocytes differentiate into 3T3 -L1 adipocytes, were observed under inverted microscope 3T3-L1 preadipocytes morphology, oil red O staining lipid droplet accumulation.2.3T3-L1 adipocytes given oleic acid and palmitic acid incubated overnight, were detected IL-6, TNF-α, MCP-1and MIP-1αsecretion by ELISE under ASP stimulation and insulin stimulation .3. 3T3-L1 adipocytes given oleic acid and palmitic acid incubated overnight, were detected JNK1 and IKKβprotein expresstion by Western Bolt under ASP stimulation and insulin stimulation .Results1.3T3-L1 preadipocytes showed a fibroblast-like spindle cells, no lipid droplets in the cytoplasm; After differentiation and maturation, the cells became round, larger, can be seen clearly in the lipid droplets around the nucleus, showing a typical ring shape.2. After incubate with ASP, comparing with that of 0 mmol/L, the levels of IL-6, TNF-α, MCP-1, MIP-1αprotein expression were dropped with increasing FFA concentrations. ASP can inhibit IL-6(P<0.05,1.0 mmol/L ), TNF-α(P<0.05,1.0 mmol/L ) , MCP-1, MIP-1α(P<0.05,1.0 mmol/L ) secretion to a certain extent . High levels of FFA can significantly inhibit JNK1 activity(P<0.05,1.0 mmol/L), restrain IKKβactivity.3. In insulin stimulating condition, comparing with the 0 mmol/L, the levels of IL-6, TNF-α(P<0.05,1.0 mmol/L), MCP-1(P<0.05,1.0 mmol/L) protein expression were dropped with increasing FFA concentrations. High levels of FFA can significantly enhance MIP - 1α(P<0.05,1.0 mmol/L) expression. With ASP different is, high levels of FFA (both oleate and palmitate) do nothing on JNK1 and IKKβexpression.ConclusionASP showed a function not only on inflammatory factors secretion in adipocytes, but also some signal molecules changes in JNK, IKK/NF-κB signal pathway , ASP may be involved in mediate inflammatory of fat cells.