罗格列酮抑制糖基化终产物诱导大鼠血管平滑肌细胞钙化机制

Mechanism of rosiglitazone on advanced glycation end products induced calcification in rat vascular smooth muscle cells

目的探讨罗格列酮抑制糖基化终产物(AGEs)诱导大鼠血管平滑肌细胞(VSMCs)钙化的可能机制。方法在含10 mmol·L^(-1)β-甘油磷酸培养液中加入200 mg·L^(-1)的AGEs及不同浓度(10^(-7)～10^(-5)mol·L^(-1))的罗格列酮体外培养VSMCs,采用甲-酚酞络合酮方法测定细胞钙含量,real time-PCR检测肿瘤坏死因子α(Tnf-α)、白细胞介素6(Il-6)及糖基化终产物受体(Rage)mRNA表达,Western blotting检测Rage蛋白表达,ELISA法检测细胞上清Tnf-α、Il-6水平。结果 AGEs可使VSMCs钙含量增加,Tnf-α、Il-6及Rage mRNA和蛋白表达增加(P<0.01);罗格列酮组均可降低VSMCs细胞层钙含量,抑制上述影响(P<0.05)。结论罗格列酮可能通过抑制VSMCs Rage表达,降低Tnf-α、Il-6表达和分泌,减轻AGEs诱导的VSMCs钙化。

AIM To investigate the mechanism of rosiglitazone on calcification in cultured rat vascular smooth muscle cells（VSMCs） by advanced glycation end products（AGEs）.METHODS VSMCs were incubated with DMEM containing 10 mmol·L^（-1） 3-glvcerophosphate and 200 mg·L^（-1） AGEs with or without 10^（-7） - 10^（-5） mol·L^（-1） rosiglitazone.Calcium deposition,the mRNA and protein expression of Tnf-α,Il-6 and Rage were determined by real time-PCR and Western blotting.RESULTS Compared with AGEs treatment alone,addition of rosiglitazone decreased calcium deposition in VSMCs at 72 h.Rosiglitazone also down-regulated AGEs-induced Rage mRNA and protein expression by about 50%at 24 h.In addition,rosiglitazone decreased both mRNA and protein expression of Tnf-α,Il-6 in VSMCs induced by AGEs at 24 h.CONCLUSION Rage contributes at least partially to the AGEs-induced calcification.Rosiglitazone might exert anticalcification effects via blockade of AGE-Rage interaction.