可溶性环氧化物水解酶抑制剂t-AUCB对巨噬细胞脂质代谢的影响

目的观察可溶性环氧化物水解酶抑制剂t-AUCB对小鼠巨噬细胞脂质摄取及降解的影响,并探明其可能机制。方法培养小鼠巨噬细胞RAW264.7,分别用不同浓度t-AUCB(1、10、50及100μmol/L)干预24 h,或在100μmol/L t-AUCB干预前1 h加入PPARγ拮抗剂GW9662 5μmol/L,采用t-AUCB 0μmol/L干预组作为空白对照。采用125Ⅰ-ox-LDL放射配基法测定小鼠巨噬细胞对氧化型低密度脂蛋白(ox-LDL)的摄取及降解,实时荧光定量PCR和Western blot分别测定小鼠巨噬细胞CD36 mRNA和蛋白表达。结果 t-AUCB呈剂量依赖性地增加巨噬细胞125Ⅰ-ox-LDL摄取量和降解量,0、1、10、50和100μmol/L t-AUCB干预时,摄取量分别为414.96±46.71μg/g、519.54±47.7μg/g、629.04±37.97μg/g、720.66±48.58μg/g和881.57±68.44μg/g,降解量分别为16180.23±967.28μg/g、17369.62±478.34μg/g、21794.85±689.36μg/g、27883.03±712.25μg/g和30194.61±635.71μg/g,与空白对照组比较差异显著(P<0.05),加入GW9662后100μmol/L组摄取量降至467.80±51.98μg/g,降解量降至16326.19±735.95μg/g,与单独加入t-AUCB组比较差异具有显著性(P<0.05);t-AUCB可呈剂量依赖性的增加小鼠巨噬细胞CD36 mRNA和蛋白的表达,而加入GW9662后明显抑制上述作用。结论 t-AUCB可通过上调PPARγ-CD36信号通路分子表达增加小鼠巨噬细胞摄取和降解ox-LDL。

Effects of Soluble Epoxide Hydrolase Inhibitors t-AUCB on Lipid Metabolism in Mouse Macrophage

Aim To observe the effects of soluble epoxide hydrolase inhibitors t-AUCB on the uptake and degradation of lipid in mouse macrophage. Methods RAW264.7 mouse macrophage was cultured,then t-AUCB in various concentration(1,10,50 and 100 μmol/L) were added for 24 hours,or incubated with peroxisome proliferators activated receptor gamma(PPARγ) antagonist GW9662(5 μmol/L).0 μmol/L t-AUCB treated group was taken as empty control.After then,the uptake and degradation of oxidized low density lipoprotein(ox-LDL) in cells were detected by radioligand assay.The mRNA and protein expression of CD36 were determined by real-time PCR and Western blot. Results t-AUCB could dose-dependently increase the uptake and degradation of ox-LDL in mouse macrophage.After stimulated with 0,1,10,50 and 100 μmol/L t-AUCB,the uptake level of ox-LDL were 414.96±46.71 μg/g,519.54±47.7 μg/g,629.04±37.97 μg/g,720.66± 48.58 μg/g,881.57±68.44 μg/g,and the degradation of ox-LDL were 16180.23±967.28 μg/g,17369.62±478.34 μg/g,21794.85±689.36 μg/g,27883.03±712.25 μg/g,30194.61±635.71 μg/g.However,after incubation of GW9662,the uptake and degradation of ox-LDL with 100 μmol/L t-AUCB were decreased to 467.80±51.98 μg/g and 16326.19±735.95 μg/g.Respectively,the difference was statistically significant(P<0.05).t-AUCB could dose-dependently increase the mRNA and protein expression of CD36,but after added with GW9662,above-mentioned function was significantly attenuated. Conclusion t-AUCB could upregulate the uptake and degradation of ox-LDL in mouse macrophage through PPARγ-CD36 pathway.