Exendin-4对猪髋动脉内皮细胞NO释放的影响.docVIP

Exendin-4对猪髋动脉内皮细胞NO释放的影响 刘浩宇，胡小春*，张冬颖，覃 数（400016重庆，重庆医科大学附属第一医院心血管内科） [摘要]目的 观察Exendin-4（Ex-4）对猪髋动脉内皮细胞（pig iliac endothelium cell lines，PIEC）释放一氧化氮（NO）的影响，及其与细胞内外钙浓度的关系。方法 免疫荧光法检测确认PIEC上存在胰高血糖素样肽-1受体（glucagon-like peptide-1，GLP-1R）；Griess重氮化反应检测Ex-4对有钙、无钙两种培养基条件下内皮细胞NO释放的影响，流式细胞仪检测细胞内钙浓度，评价Ex-4诱导PIEC释放NO的时效-量效关系。给予内皮型一氧化氮合成酶（eNOS）信号阻断剂左旋-硝基精氨酸甲脂（L-NAME）预处理后，观察Ex-4对PIEC释放NO的影响。结果 PIEC上有GLP-1受体表达。Ex-4能刺激PIEC释放NO，作用有剂量及时间依赖关系，同时细胞内钙离子浓度也有升高[（62.67±1.53）vs（57.36±1.42），P＜0.01]。无钙培养液组PIEC释放NO较有钙培养液组升高更明显[（2.014±1.040）μmol/L vs （1.624±0.424）μmol/L，P=0.03]。Ex-4刺激PIEC释放NO L-NAME预处理后，Ex-4诱导的PIEC释放NO减少（P＜0.01）。结论 Ex-4通过激活eNOS增加内皮细胞NO释放，其eNOS激活作用可能与细胞内外钙离子变化相关。 [关键词]胰高血糖素样肽-1；一氧化氮；内皮细胞 Effects of Exendin-4 on NO production in the pig iliac endothelium LIU Haoyu， HU Xiaochun，ZHANG Dongying，QIN Shu（Department of Cardiology， the First Affiliated Hospital of Chongqing Medical University，Chongqing 400016， China） Abstract： Objective To investigate the effects of Exendin-4 （Ex-4） on nitric oxide （NO） produced by pig iliac endothelium cells （PIEC） and their relationship under different conditions of intracellular/extracellular calcium， and explore the mechanism. Methods PIEC were cultured in calcium and non calcium nutrient medium separately. GLP-1 receptor （GLP-1R） was studied using immunofluorescence. PIEC were treated with various concentrations of Ex-4. The NO production was measured using Griess Reaction in cell culture supernatants. The intracellular calcium concentration was measured using Flow Cytometer with the fluorescence probe Fluo-3 AM. Results GLP-1R was expressed in the PIEC. Ex-4 increased the PIEC NO production in a dose and time dependent manner， and increased the intracellular calcium concentration accordingly(MFI 62.67±1.53 vs MFI 57.36±1.42，P＜0.01）. NO production in the pig iliac endothelium cell cultured with non calcium nutrient medium was higher compared with those in calcium nutrient medium(2.014±1.040 μmol/l vs 1.624±0.424，P=0.03）. The NO production