替米沙坦对AGEs诱导人内皮细胞表达VCAM-1及MCP-1的作用和机制

摘要

Objective To investigate the capacity of telmisartan in inflammation induced by advanced glycation end products (AGEs) and discuss the proposed mechanism in vitro.Methods Human umbilical vein endothelial cells (HUVECs) were prepared by collagenase digestion and cultured in vitro. The reactiveoxygen species (ROS) level in HUVECS was observedthrough inversion fluoescence microscope and detected by ROS detection kit. An increase in VCAM-1, MCP-1 and receptor for AGEs(RAGE) mRNA was analyzed by reverse transcription-polymerase chain reaction (RT-PCR) as markers of AGEs -induced endothelial inflammation.Results Compared with control group,the fluorscence intensity of ROS in AGEs group was stronger.Telmisartan could decline the ROS level induced by AGEs obviously in HUVECs.AGEs significantly increased the levels of VCAM-1 mRNA, MCP-1mRNA concentration -dependently in HUVECs.AGEs at the dose of 10-4 mg/mlwas able toaugment the expression of VCAM-1 and MCP-1 mRNA significantlycompared with the control group（0.24±0.01 vs 0.07±0.02; 0.25±0.01 vs 0.18±0.03, P＜0.05).These effects were significantly attenuated by pretreatment with telmisartan.Telmisartan (10 nmol/L)markedly reduced VCAM-1 mRNA and MCP-1mRNA expression stimulated by AGEs compared with the AGEs group（0.23±0.01 vs 0.85±0.11; 0.62±0.10 vs 0.1.05±0.04, P＜0.05); Furthermore,telmisartan at the dose of 1nmol/lmarkedlyinhibited RAGE mRNA expression caused by AGEs（0.64±0.03 vs 1.18±0.10, P＜0.05).Conclusion AGEs could significantly increase the expression of VCAM-1 and MCP-1 in HUVECs. Telmisartan exerts potent anti-inflammatory effects on endothelial cells,possiblyresting on lowering RAGE mRNA level.%目的 通过观察替米沙坦对晚期糖基化终末产物(AGEs)诱导的人脐静脉内皮细胞表达血管细胞黏附因子-1(VCAM-1)和单核细胞趋化蛋白-1(MCP-1)的影响,研究替米沙坦对AGEs所致动脉粥样硬化(AS)的干预作用和机制.方法 采用胶原酶消化法获取人脐静脉内皮细胞,分为4组:空白对照组,牛血清白蛋白(BSA)对照组,AGEs诱导组(10-4~10-1mg/mL),AGEs+替米沙坦组(1、10、100 nmol/L).活性氧检测试剂盒检测及倒置荧光显微镜观察细胞内活性氧含量,RT-PCR检测VCAM-1、MCP-1及晚期糖基化终末产物受体(RAGE)的mRNA.结果 AGEs组人内皮细胞内活性氧荧光强度增强,替米沙坦干预后降低;AGEs呈浓度依赖性地增强人内皮细胞对VCAM-1和MCP-1基因的转录,与空白对照组相比,AGEs(10-4mg/mL)组VCAM-1和MCP-1 mRNA表达水平显著增高(0.24±0.01 vs 0.07±0.02;0.25±0.01 vs 0.18±0.03,P<0.05);替米沙坦呈浓度依赖性地抑制人内皮细胞对VCAM-1和MCP-1基因的转录,与AGEs诱导组相比,替米沙坦(10 nmol/L)组人内皮细胞的VCAM-1和MCP-1基因转录水平显著降低(0.23±0.01 vs 0.85±0.11;0.62±0.10 vs 1.05±0.04,P<0.05);与AGEs诱导组相比,替米沙坦(1 nmol/L)组人内皮细胞RAGE基因表达水平显著降低(0.64±0.03 vs 1.18±0.10,P<0.05).结论 AGEs增强人内皮细胞表达VCAM-1和MCP-1;替米沙坦可能通过抑制RAGE表达来抑制AGEs诱导的人内皮炎性损伤.