高糖依赖过氧化体增殖物激活型受体γ介导THP-1巨噬细胞CD36表达及脂质蓄积

High Glucose Induces CD36 Expression and Lipid Accumulation Through Peroxisome Proliferator Activated Receptor γ in THP-1 Macrophages

目的观察高糖诱导THP-1巨噬细胞CD36表达及脂质蓄积是否由过氧化体增殖物激活型受体γ(PPARγ)所介导。方法分别用20 mmol/L D-葡萄糖(高糖)、高糖+10 mmol/L GW9662(PPARγ拮抗剂)、50mg/L氧化型低密度脂蛋白+高糖、50 mg/L氧化型低密度脂蛋白+高糖+10 mmol/L GW9662共同孵育THP-1巨噬细胞24 h;采用逆转录聚合酶链反应检测CD36、PPARγmRNA的表达;用Western blot分别检测CD36、PPARγ蛋白质的表达;用油红O染色观测细胞内脂质蓄积情况,高效液相色谱分析法检测细胞内总胆固醇水平。结果GW9662明显抑制高糖所诱导的THP-1巨噬细胞CD36的表达及脂质蓄积,CD36 mRNA和蛋白质表达明显下降(P<0.05);细胞内脂滴明显减少,总胆固醇含量明显下降(P<0.05)。结论高糖所诱导的CD36表达及脂质蓄积可能是由PPARγ所介导的。本研究将为糖尿病性动脉粥样硬化病变的防治积累有价值的实验资料。

Aim To investigate the effect of peroxisome proliferator activated receptor γ（PPARγ） on the expression of CD36 and the lipid accumulation induced by high glucose in THP-1 macrophages. Methods THP-1 macrophages were incubated with 20 mmol /L D-glucose,10 mmol /L GW9662,and 50 mg /L oxidized low density lipoprotein（ox-LDL） for 24 h,combinated or respectively. The total cholesterol contents in THP-1 macrophages were determined by high performance liquid chromatography,and the lipid accumulation was detected by oil red O stain. CD36 and PPARγmRNA level were determined by reverse transcription polymerase chain reaction（ RT-PCR）,respectively. CD36 and PPARγ protein level were determined by Western blot. Results GW9662 significantly inhibited the expression of CD36 and lipid accumulation induced by high glucose in THP-1 macrophages. The expression of CD36 mRNA and protein was significantly suppressed by GW9662（ P 〈 0. 05）,intracellular lipid decreased obviously and the total cholesterol in THP-1 macrophages was markedly reduced subsequently（ P 〈 0. 05）. Conclusion High glucose induces CD36 expression and lipid accumulation through the modulation of PPARγ in THP-1 macrophages.