CAJ | 학술논문

目的观察可溶性环氧化合物水解酶抑制剂(sEHi)tAUCB对脂肪细胞胆固醇流出的影响,并探讨其机制.方法测定脂肪细胞过氧化物酶体增殖物激活受体γ(PPARγ)及三磷酸腺苷结合盒转运体A1(ABCA1)mRNA及蛋白的表达,检测细胞内胆固醇流出.结果 tAUCB呈剂量依赖性促进载脂蛋白(Apo)A1介导的胆固醇流出,1、10、50、100 μmol/L浓度的tAUCB干预后,胆固醇流出率分别为(5.93±0.66)%,(7.40±0.43)%,(8.30±0.34)%和(9.77±0.42)%.加入10～100μmol/L tAUCB干预组与空白组(5.67±0.17)%比较,胆固醇流出差异有统计学意义(P＜0.05).同时发现随着tAUCB干预浓度的增加,脂肪细胞ABCA1、PPARy mRNA及蛋白的表达也呈剂量依赖性地升高,而GW9662明显抑制tAUCB对脂肪细胞胆固醇流出及ABCA1和PPARγ表达的促进作用.结论 tAUCB可通过上调PPARy的表达,促进脂肪细胞Apo A1-ABCA1通路加速细胞内胆固醇流出,抑制脂肪细胞内胆固醇过度蓄积.
목적 관찰가용성배양화합물수해매억제제(sEHi)tAUCB대지방세포담고순류출적영향,병탐토기궤제.방법 측정지방세포과양화물매체증식물격활수체γ(PPARγ)급삼린산선감결합합전운체A1(ABCA1)mRNA급단백적표체,검측세포내담고순류출.결과 tAUCB정제량의뢰성촉진재지단백(Apo)A1개도적담고순류출,1、10、50、100 μmol/L농도적tAUCB간예후,담고순류출솔분별위(5.93±0.66)%,(7.40±0.43)%,(8.30±0.34)%화(9.77±0.42)%.가입10～100μmol/L tAUCB간예조여공백조(5.67±0.17)%비교,담고순류출차이유통계학의의(P＜0.05).동시발현수착tAUCB간예농도적증가,지방세포ABCA1、PPARy mRNA급단백적표체야정제량의뢰성지승고,이GW9662명현억제tAUCB대지방세포담고순류출급ABCA1화PPARγ표체적촉진작용.결론 tAUCB가통과상조PPARy적표체,촉진지방세포Apo A1-ABCA1통로가속세포내담고순류출,억제지방세포내담고순과도축적.
Objective To observe the effects of soluble epoxide hydrolase inhibitors tAUCB on cholesterol efflux in adipocytes. Methods 3T3-L1 preadipocytes were induced to differentiation and maturation. Cells were stimilated with 100μg/L LPS after starved for 24 hours, then tAUCB in various concentrations(1 ,10,50,100 μmol/L)were added for 24 h, or incubated with the peroxisome proliferator activated receptor gamma (PPARy) antagonist GW9662 (5 μmol/L).0μmol/L tAUCB treated group was taken as empty control. After then, the mRNA expression of PPARγ and adenosine triphosphate binding cassette transporter Al (ABCA1) in cells were determined via realtime-PCR, the amounts of protein expression of PPARγand ABCA1 in cells were detected by Western blot, the efflux rates of 3H-cholesterol in cells were detected by means of liquid scintillation counter. Results tAUCB could dose-dependently increase the apolipoprotein A1 (apoA1)-mediated cholesterol efflux in adipocytes. After stimulated by 1, 10,50,100 μmol/L tAUCB, cholesterol efflux rates were (5.93±0.66) %, (7.40 ± 0. 43) %, (8. 30 ±0. 34)% ,(9.77±0.42)% respectively, there were significant difference after treated by 10-100 μmol/L tAUCB compared with control(5.67±0.17)%(P＜0.05). With the concentration of tAUCB increased,ABCA1, PPAR mRNA and protein expression were also dose-dependently up-regulated. GW9662 could significantly inhibit the effects of tAUCB, and then reduce the cholesterol efflux and the expression of PPARγ and ABCA1 in adipocytes. Conclusions tAUCB could up-regulate PPARγ expression in adipocytes, and promote the cholesterol efflux of adipocytes via apoA1-ABCA1 pathway, which might decrease the cellular cholesterol accumulation in adipocytes.