摘要
目的:建立一种快速简便测定神经元中游离的花生四烯酸的方法,分析培养的神经元在热处理前后细胞和细胞膜中花生四烯酸的水平变化,为探讨热环境对神经元损伤提供参考。方法:原代培养的成熟的大鼠纹状体神经元经过43℃处理1h,细胞裂解后提取细胞裂解液和细胞膜中的脂肪酸,加入C17:0作为内标,甲醇酯化后用气相色谱和质谱联用的方法,测定脂肪酸甲酯的水平。结果:在细胞中含量相对丰富的饱和脂肪酸(棕榈酸和硬脂酸)水平,以及不饱和脂肪酸(油酸、亚油酸和花生四烯酸)水平在热处理前后没有明显改变,而细胞质中花生四烯酸水平在热处理后明显升高(0.53±0.13比1.00±0.56,F=5.71,P<0.05),升幅达到100%。同时在所分析的饱和脂肪酸和不饱和脂肪酸水平在在热处理前后没有显著差别,而细胞膜中花生四烯酸在热处理前后均检测不到。结论:热环境处理导致纹状体神经元更多的花生四烯酸释放,可能介导热对神经元的损伤。
AIM:To setup a convenient method for the measurement of free arachidonic acid(AA) in neurons and to analyze the effects of heat treatment on the AA levels in neurons so as to give a reference to the study on heat injury in neural system.METHODS:Primary cultured striatum neurons of rats were treated at a temperature of 43 ℃for 1 h.Cells of the heat treated group and control group were homogenized,and the fatty acids in cell lysate and cell membrane were extracted respectively.After heptadecanoic acid(C17:0) was added as an internal standard,the levels of fatty acid methanol esters were detected with Gas/Mass Chromatography.RESULTS:The relative abundant fatty acid,including saturated(palmitic acid and stearic acid) and unsaturated(oleic acid,linoleic acid and arachidonic acid) were detectable,and did not change significantly during heat treatment,either in cell lysate or in cell membrane.There were no significant difference between saturated acid and unsaturated acid.While the level of AA in cytoplasm increased 100%from 0.53±0.13 to 1.00±0.56 after heat treatment(F=5.71,P< 0.05),but the level of AA in cell membrane was undetectable,even after heat treatment.CONCLUSION:More amount of AA is released from neurons in striatum induced by heat treatment, and the increased AA may be crucial in the injury signal transduction.
出处
《中国临床康复》
CSCD
2004年第7期1262-1263,共2页
Chinese Journal of Clinical Rehabilitation
