吡格列酮对高糖诱导的小鼠主动脉平滑肌细胞晚期糖基化终末产物受体表达的影响及机制

Effects of pioglitazone on advanced glycation end products receptor expression in hyperglycemicinduced aortic vascular smooth muscle cells in mice and its mechanism

目的探讨吡格列酮(PIO)对高糖诱导的小鼠主动脉血管平滑肌细胞(VSMCs)晚期糖基化终末产物受体(RAGE)表达的影响及其作用机制。方法原代培养小鼠主动脉VSMCs分为5组:(1)5. 5 mmol/L葡萄糖(正常浓度葡萄糖组,NG);(2)25 mmol/L葡萄糖(高浓度葡萄糖组,HG);(3)25 mmol/L葡萄糖+10μmol/L PIO(HG+PIO);(4)25 mmol/L葡萄糖+10μmol/L PIO+10μmol/L GW9662 (HG+PIO+GW9662);(5) 25 mmol/L葡萄糖+10μmol/L GW9662 (HG+GW9662)。应用RT-PCR及Western blot检测VSMCs中RAGE及过氧化物酶体增殖物激活受体γ(PPAR-γ)的表达。结果高糖增加VSMCs中RAGE的mRNA(P=0. 001)和蛋白(P=0. 002)表达;PPAR-γ激动剂PIO可抑制高糖增加VSMCs中RAGE的mRNA(P=0. 014)和蛋白(P=0. 006)表达的作用;加用PPAR-γ抑制剂GW9662后可明显减弱PIO对RAGE的mRNA(P=0. 001)和蛋白(P=0. 004)表达的抑制作用。结论 PIO可通过激活PPAR-γ,抑制高糖诱导的小鼠主动脉平滑肌细胞RAGE的表达。

Objective To analyze the effect of pioglitazone on the expression of the receptor of advanced glycation end products （RAGE） in hyperglycemic-induced mouse aortic vascular smooth muscle cells （VSMCs） and its mechanism of action. Methods The cultured primary mouse aortic VSMCs were divided into five groups： （1） 5.5 mmol/L D-glucose （normal glucose, NG）; （2） 25 mmol/L D-glucose （high glucose, HG）; （3） 25 mmol/L D-glucose with 10 μmol/L PIO （HG＋PIO）; （4） 25 mmol/L D-glucose with 10 μmol/L PIO and 10 μmol/L GW9662 （HG＋PIO＋GW9662）; （5） 25 mmol/L D-glucose with 10 μmol/L GW9662 （HG＋GW9662）. Western blot and RT-PCR were used to determine the expression of RAGE and PPAR-γ in VSMCs. Results In cultured VSMCs, high glucose increases the RAGE mRNA （ P =0.001） and protein （ P =0.002） expression, and PIO downregulated RAGE mRNA （ P =0.014） and protein （ P =0.006） expression. Blockade of PPAR-γ activity by GW9662 significantly attenuated the inhibitory actions of PIO in cultured VSMCs with high-glucose challenge on RAGE mRNA （ P =0.001） and protein （ P =0.004）. Conclusions PIO has been shown to inhibit high glucose-induced RAGE expression in mouse aortic VSMCs by activating PPAR-γ.