脂筏调控不同饱和度反式脂肪酸诱导内皮细胞凋亡信号转导通路研究

Hydrogenated oils and high temperature heating oils containing a large amount of of trans fatty acids (TFA).TFA have a serious impact on human health. It is reported that TFA can induce endothelial apoptosis, but the specific mechanism is not clear. As a platform, lipid rafts were reported to participate in intracellular apoptotic signal pathway. But the trans fatty acids can induce apoptosis in endothelial cells through lipid rafts has not been reported.The research induce endothelial cells with different saturation trans fatty acids, combined with two-dimensional electrophoresis, screened apoptosis pathway protein mediated by lipid rafts and defined the pathway. Through changing the cholesterol structure of lipid rafts, detect the protein level of apoptoic molecule and co-localization of lipid rafts. Ultimately determine the signal transduction pathway with lipid raft-dependent correlation. The mechanism of apoptoic signal pathways mediated by lipid rafts playing an important theoretical value for understanding the TFA-induced apoptosis progress and the relation between lipid rafts.

很多研究证明反式脂肪酸对人体健康造成严重影响。研究证实反式脂肪酸（trans fatty acid, TFA）能诱导内皮细胞凋亡，但目前作用机制不明确。研究报道脂筏作为信号分子聚集的平台，参与调控了细胞凋亡信号转导通路。但目前反式脂肪酸能否通过脂筏诱导内皮细胞凋亡尚无报道。本研究以脂筏为靶点，采用双向电泳及质谱分析筛选由脂筏调控的凋亡通路标志蛋白，确定凋亡通路；借助脂筏胆固醇干扰，采用流式细胞术检测凋亡通路蛋白表达差异变化，并结合激光共聚焦显微镜观察通路标志蛋白与脂筏共定位，阐明凋亡信号通路与脂筏依赖关系机制。本研究为深入理解脂筏在反式脂肪酸诱导细胞凋亡过程中的调控作用提供理论依据。

很多研究证明反式脂肪酸对人体健康造成严重影响。研究证实反式脂肪酸（trans fatty acid; TFA）能诱导内皮细胞凋亡，但目前作用机制不明确。研究报道脂筏作为信号分子聚集的平 台，参与调控了细胞凋亡信号转导通路。但目前反式脂肪酸能否通过脂筏诱导内皮细胞凋亡尚无报道。本研究成功建立了脂筏提取和脂筏胆固醇干扰技术。通过蛋白组学共获得可信蛋白总数：4741个，其中动态变化中差异表达蛋白数目174个，筛选到差异信号通路103条，通过转录组学筛选出差异表达RNA221条，筛选到差异信号通路192条。通过western验证了包括AMPK、HIF-1等与脂筏相关的凋亡信号转导通路，并通过脂筏胆固醇干扰，借助免疫荧光观测了FAS与脂筏共定位变化，确定了脂筏与FAS相关死亡受体凋亡信号通路呈依赖关系。以脂筏作为研究靶点，通过揭示脂筏参与调控不同饱和度TFA诱导内皮细胞凋亡的作用机制，为研究脂筏在TFA诱导内皮细胞凋亡过程中的作用提供理论依据，对于研究反式脂肪酸与人类营养健康具有重要意义。