游离脂肪酸及葡萄糖对鼠主动脉内皮细胞影响及作用机制.docVIP

游离脂肪酸及葡萄糖对鼠主动脉内皮细胞影响及作用机制 　　[摘要] 目的 探讨高糖高游离脂肪酸（free fatty acids，FFAs）对内皮细胞增殖和凋亡的影响及其作用机制。 方法 原代培养大鼠主动脉内皮细胞（rat aorta endothelial cell，RAEC），采用不同浓度的葡萄糖（5.5 mmol/L，30 mmol/L）及棕榈酸（200 μmol/L、400 μmol/L、600 μmol/L）单独或联合作用于细胞24 h、48 h、72 h。免疫组织化学染色方法鉴定内皮细胞并测定IL-8、Bcl-2、BAX表达水平；MTT比色法检测细胞增殖率。 结果 FFAs及葡萄糖单独及联合应用均可导致细胞出现凋亡形态学变化，增殖呈剂量-时间依赖性（P ＜ 0.05），且联合组明显低于单独培养组（P ＜ 0.01）；干预后IL-8、BAX表达增加，Bcl-2蛋白表达逐渐减弱，Bcl-2/BAX比值逐渐减小，联合培养组表达更为明显。 结论 高浓度FFAs及高糖具有抑制内皮细胞生长、促进其凋亡的作用，其作用机制可能是通过葡萄糖及棕榈酸酯参与PI3K/AKT等途径激活氧化应激诱导细胞凋亡。 　　[关键词] 糖尿病；游离脂肪酸；内皮细胞；bcl-2；BAX；PI3K/AKT 　　[中图分类号] R34[文献标识码] A[文章编号] 1673-9701（2012）10-0004-04 　　 　　Influence and mechanism of free fatty acid and glucose on endothelial cell of rat aorta 　　GUO Hailian1 LIU Xiaoling2 LI Pengcui3 HAN Le2 WANG Dongqing1 　　1.Graduate School of Shanxi Medical University, Taiyuan 030001, China; 2.Department of Endocrinology, the Second Affiliated Hospital of Shanxi Medical University, Taiyuan 030001，China； 3.Department of Orthopaedic Laboratory, the Second Affiliated Hospital of Shanxi Medical University, Taiyuan 030001, China 　　[Abstract] Objective To explore the influence and mechanism of free fatty acid（FFAs） and glucose on endothelial cell proliferation and apoptosis. Methods Rat aorta endothelial cell （RAEC）were cultured in vitro, glucose(5.5 mmol/L, 30 mmol/L) and palmitic acid (200 μmol/L、400 μmol/L、600 μmol/L) in different concentration were applied，alone and together, to RAEC for 24 h， 48 h， 72 h. Immunohistochemistry were used to determine CD31-related antigen and detectedIL-8, Bcl-2, BAX protein expression changes; Applied four methyl azo thiazole blue (determined by MTT) colorimetric detection different environment RAEC proliferation capacity. Results Glucose and FFAs, alone and together, can lead to apoptotic morphological changes in RAEC； FFAs and glucose display strong growth inhibitory effect in a-time and does-development manner against RAEC (P 　　1.2 主动脉内皮细胞的培养与鉴定 　　颈椎脱臼处死SD雌性大鼠，碘伏酒精消毒后，逐层剖开胸腹腔，于脊柱旁暴露主动