间歇性低氧对肝细胞脂代谢的影响及其可能机制的-第三军医大学学报.docVIP

间歇性低氧对肝细胞脂代谢的影响及其可能机制的探索 孙中新，沈 薇 （400010 重庆，重庆医科大学附属第二医院消化内科） [摘要] 目的 探讨间歇性低氧对肝细胞脂质代谢的影响。方法 利用三气培养箱及两珠肝细胞L02 HepG2建立间歇性低氧细胞模型，经甘油三酯（TG）测定肝细胞发生脂肪性变后，荧光显微镜观察间歇性低氧对肝细胞内ROS的影响，Western blot检测ROS对HIF-1α、HIF-2α蛋白表达的调节，western blot检测HIF-1α下游与脂质代谢相关的SREBP-1c、FAS蛋白的表达。结果 实验组较对照组肝细胞内出现脂质异常沉积，经间歇性低氧处理5d后L02 HepG2肝细胞TG含量较对照组明显增多分别为（78.7±2.73）vs（25.11±5.86）,t=16.58 P0.05; （96.4±3.67）vs（21.4±3.9）,t=28.01, P0.05;表明肝细胞间歇性低氧肝细胞脂变模型构建成功。ROS在对照组有基础表达，随着间歇性低氧处理时间延长L02 HepG2细胞内ROS的含量增多绿色荧光琢渐增强。Western blot结果显示实验组较对照组 HIF-1α、SREBP-1c 、FAS 表达增加并且随时间的延长表达量增多（P0.05），受间歇性低氧的影响4d、5d后，HIF-2α的蛋白表达较对照组明显减少（P0.05）。结论 间歇性低氧诱导产生的ROS通过调节低氧诱导因子和脂肪分化相关蛋白的表达，影响HIF-1α-SREBP-1c-FAS途径导致肝细胞脂代谢异常。 [关键词] 间歇性低氧；活性氧；肝细胞脂变；低氧诱导因子 [中图法分类号] [文献标志码] A Effect of intermittent hypoxia on lipid metabolism in liver cells and its underlying mechanism Sun Zhongxin，Shen Wei，（The Second Af?liated Hospital of Chongqing Medical University，Gastroenterology，Chongqing， China，400010） [Abstract] Objective to explore the effect of internmentent hypoxia on lipid metabolism in human hepatic cell lines and its underlying mechanism. Methods Forma3131 incubator and L02cells and HepG2 cells were used to establish intermentent hypoxia cell model,after TG in hypatocyte were observed by biochemical assays. The effect of intermittent hypoxia on intracellular reactive oxygen species in hypatocyte is observed by Inverted Flurescence Microscopy.the expression of protein levels hypoxia-induced-1α(HIF-1α)、HIF-2αwhich were regulated by ROS were detected by western-blot;the protein of triaeylglyceml synthesis SREBP-1c FAS,which were the downstream of HIF-1αwere detected by western-blot Results the accumulation of lipid droplets and the content of TG in the experiment groups were significantly higher than the control groups in L02、hepG2cells, repectively（78.7±2.73）vs（25.11±5.86），t=16.58 p0.05;（96.4±3.67）vs（21.4±3.9）t=28.01, p0.05;they suggested that the intermentent hypoxia cell model is su