游离脂肪酸及葡萄糖对鼠主动脉内皮细胞的影响及作用机制(1)
[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。
, http://www.100md.com
[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
, 百拇医药
[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 < 0.05), and the combination group was significantly lower than the proliferation of cultured alone group (P < 0.01). After intervention, IL-8, BAX expression were increased, Bcl-2 protein expression were gradually decreased, Bcl-2/BAX ratio wrere decreased, and more obvirous expression of the combination group. Conclusion High FFAs and high glucose can inhibit the growth of RAEC and promote their apoptosis, its mechanism may be involved by glucose and palmitate PI3K/AKT pathway activation oxidative stress-induced apoptosis.
, 百拇医药
[Key words] Diabetes mellit; Free fatty acid; Endothelial cell; bcl-2; BAX; PI3K/AKT
糖尿病血管病变是糖尿病的主要并发症之一，也是糖尿病患者致残和致死的主要并发症，其中血管内皮细胞的损伤在血管并发症中起着关键作用[1]。2001年美国糖尿病学会年会上，BANTING科学奖得主McGarry JD 教授提出，脂代谢障碍是2型糖尿病及其并发症的基本病理生理改变[2]。伴随着葡萄糖利用和代谢障碍，脂肪的合成代谢障碍而分解代谢异常增高，血液中FFAs含量增多，而FFAs在一定程度上可反映2型糖尿病患者体内脂代谢异常状况。FFAs尤其是棕榈酸(palmitic acid，PA)可导致内皮细胞功能障碍,是糖尿病血管病变的关键因素。因此，探讨脂毒性对糖尿病血管病变的影响及可能的机制对临床具有一定的指导意义。本实验观察高糖状态下不同浓度的棕榈酸对大鼠主动脉内皮细胞的影响，并探讨其可能的作用机制，为糖尿病血管病变的防治提供新的思路。
, 百拇医药
1 材料与方法
1.1 主要材料和试剂
SD大鼠(山西医科大学实验动物中心)，软脂酸(Gibco，USA)，RPMI 1640(武汉博士德)，胎牛血清(Cyagen Biosciences，USA)，大鼠内皮细胞生长因子(VEGF)(Prospec，USA)，胰蛋白酶(TRYPSIN 1：250)(Solarbio)，去脂牛血清白蛋白(d-BSA)、四甲基偶氮唑蓝(MTT)(sigma，USA)，即用型兔抗大鼠CD31单克隆一抗试剂盒(武汉博士德)、即用型兔抗大鼠bcl-2单克隆一抗试剂盒(武汉博士德)、即用型兔抗大鼠BAX单克隆一抗试剂盒(武汉博士德)、即用型SABC免疫组织化学染色试剂盒(武汉博士德)，DAB显色液(武汉博士德)。, http://www.100md.com(郭海莲 刘晓玲 李鹏翠 韩乐 王东青)
, http://www.100md.com
[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
, 百拇医药
[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 < 0.05), and the combination group was significantly lower than the proliferation of cultured alone group (P < 0.01). After intervention, IL-8, BAX expression were increased, Bcl-2 protein expression were gradually decreased, Bcl-2/BAX ratio wrere decreased, and more obvirous expression of the combination group. Conclusion High FFAs and high glucose can inhibit the growth of RAEC and promote their apoptosis, its mechanism may be involved by glucose and palmitate PI3K/AKT pathway activation oxidative stress-induced apoptosis.
, 百拇医药
[Key words] Diabetes mellit; Free fatty acid; Endothelial cell; bcl-2; BAX; PI3K/AKT
糖尿病血管病变是糖尿病的主要并发症之一，也是糖尿病患者致残和致死的主要并发症，其中血管内皮细胞的损伤在血管并发症中起着关键作用[1]。2001年美国糖尿病学会年会上，BANTING科学奖得主McGarry JD 教授提出，脂代谢障碍是2型糖尿病及其并发症的基本病理生理改变[2]。伴随着葡萄糖利用和代谢障碍，脂肪的合成代谢障碍而分解代谢异常增高，血液中FFAs含量增多，而FFAs在一定程度上可反映2型糖尿病患者体内脂代谢异常状况。FFAs尤其是棕榈酸(palmitic acid，PA)可导致内皮细胞功能障碍,是糖尿病血管病变的关键因素。因此，探讨脂毒性对糖尿病血管病变的影响及可能的机制对临床具有一定的指导意义。本实验观察高糖状态下不同浓度的棕榈酸对大鼠主动脉内皮细胞的影响，并探讨其可能的作用机制，为糖尿病血管病变的防治提供新的思路。
, 百拇医药
1 材料与方法
1.1 主要材料和试剂
SD大鼠(山西医科大学实验动物中心)，软脂酸(Gibco，USA)，RPMI 1640(武汉博士德)，胎牛血清(Cyagen Biosciences，USA)，大鼠内皮细胞生长因子(VEGF)(Prospec，USA)，胰蛋白酶(TRYPSIN 1：250)(Solarbio)，去脂牛血清白蛋白(d-BSA)、四甲基偶氮唑蓝(MTT)(sigma，USA)，即用型兔抗大鼠CD31单克隆一抗试剂盒(武汉博士德)、即用型兔抗大鼠bcl-2单克隆一抗试剂盒(武汉博士德)、即用型兔抗大鼠BAX单克隆一抗试剂盒(武汉博士德)、即用型SABC免疫组织化学染色试剂盒(武汉博士德)，DAB显色液(武汉博士德)。, http://www.100md.com(郭海莲 刘晓玲 李鹏翠 韩乐 王东青)