The Effects Of Soluble Epoxide Hydrolase Inhibitors On Lipid Metabolism And Secretive Function Of Adipocytes

Objectives We will illustrate the relationship inflammation, adipocytes and AS for further comprehension of anti-AS by soluble epoxide hydrolase inhibitor (sEHi), which maybe help to determine a novel therapeutic target in treatment of obesity-related metabolic diseases. via investigating the effect of AEPU on the uptake and degradation of ox-LDL by inflammatory adipocytes and its involved mechanism, detecting the effect of AEPU on the cholesterol efflux in adipocytes in inflammation, observing the secretion of ADP and TNF-αby adipocytes in inflammation, exploring the effect of AEPU on the secretion of ADP and TNF-αby adipocytes and its possible mechanism.Methods 3T3-L1 adipocytes were cultured and induced to differentiation and maturation. After cells were starved for 24 hours with 100ng/ml LPS, AEPU in various concentrations(0,1,10,50,100μmol/L) were added for co-incubation for 24 hours. Or were incubated with the PPARγantagonist GW9662 (5μmol/L) as control group. After then, the uptake and degradation of ox-LDL by adipocytes in individual groups were measured by radioligand assay, the efflux rates of 3H-cholesterol in cells were detected by means of liquid scintillation counter, the amounts of total protein of CD36, PPARγand ABCA1 in cells were detected by Western blot, the mRNA expression of CD36, PPARγand ABCA1 in cells was determined via Realtime-PCR, ADP and TNF-αin supernatant was detected with ELISA.Results1. 125I-ox-LDL was uptaked by adipocytes, which was relative to the protein and mRNA expression of PPARγand CD36. In contrast, AEPU increased the uptake of 125I-ox-LDL by inflammatory adipocytes via the pathway of PPARγ-CD36.2. LPS decreased cholesterol efflux in stimulating adipocytes, while AEPU promoted the cholesterol efflux from inflammatory adipocytes in via the pathway of PPARγ-ABCA1, Which inhibit the accumulation of the cholesterol in adipocytes.3. AEPU decreased the levels of TNF-αin inflammatory adipocytes dose-dependently and increased the levels of ADP.Conclusions1. Inflammation can impair the ability of adipocytes'uptaking ox-LDL while AEPU can enhance ox-LDL uptake by inflammatory adipocytes through PPARγ-CD36 pathway.2. Inflammation can decrease the cholesterol efflux of adipocytes while AEPU can promote the cholesterol efflux of inflammatory adipocytes via PPARγ-ABCA1 pathway.3. LPS can induce the inflammatory response of adipocytes via increasing the secretion of TNF-αfrom adipocytes while AEPU can inhibit TNF-αsecretion and upgrade the levels of ADP in inflammatory adipocytes through the up-regulation of PPARy protein and mRNA expression, which maybe one of anti-inflammatory mechanisms by AEPU.