Sasaki T, Asano T, Takakura K, Sano K, Nakamura T, Suzuki N, Imabayashi S, Ishikawa Y
No To Shinkei. 1982 Dec;34(12):1191-6.
The relationship between lipid peroxides in cerebrospinal fluid (CSF) and the occurrence of cerebral vasospasm following subarachnoid hemorrhage (SAH) was evaluated by analyzing CSF with high-performance liquid chromatography (HPLC) and gas chromatography mass spectrometry (GC-MS). Hydroperoxy eicosatetraenoic acids (HPETEs) and hydroxy eicosatetraenoic acids (HETEs) were synthesized by the treatment of arachidonic acid with hydrogen peroxide and cupric chloride. The retention time of these HPETEs and HETEs were determined on HPLC. The position of oxydation occurred was determined after methylation, reduction and trimethyl silylation using GC-MS. Thus the elucidation of positional isomers of HPETEs and HETEs was made possible by the retention time on HPLC. The supernant of CSF after SAH was adjusted to pH 3.0 and then absorbed to octadecyl silyl silica column. The eluted fraction with 15% ethanol-water from octadecyl silyl silica column was analyzed by HPLC detecting at 238 nm. No peak was observed on HPLC at the region of HPETEs and HETEs in the CSF obtained from healthy person. In SAH patients, several peaks were recognized in accordance with the occurrence of cerebral vasospasm. One of the peaks was identified as 5-HETE by HPLC and GC-MS. In 10 SAH patients, semi-quantitative analysis of 5-HETE in the CSF was performed by measuring the height of the peak identified as 5-HETE on HPLC. The close correlation was recognized between the occurrence of cerebral vasospasm and the appearance of 5-HETE in the CSF. The results of the present study suggest that lipid peroxidation is involved in the pathogenesis of chronic vasospasm after SAH.
通过高效液相色谱法(HPLC)和气相色谱 - 质谱联用仪(GC - MS)分析脑脊液(CSF),评估蛛网膜下腔出血(SAH)后脑脊液中脂质过氧化物与脑血管痉挛发生之间的关系。用过氧化氢和氯化铜处理花生四烯酸合成氢过氧化二十碳四烯酸(HPETEs)和羟基二十碳四烯酸(HETEs)。在HPLC上测定这些HPETEs和HETEs的保留时间。使用GC - MS在甲基化、还原和三甲基硅烷化后确定氧化发生的位置。因此,通过HPLC上的保留时间可以阐明HPETEs和HETEs的位置异构体。SAH后脑脊液上清液调节至pH 3.0,然后吸附到十八烷基硅烷硅胶柱上。用15%乙醇水溶液从十八烷基硅烷硅胶柱上洗脱的馏分通过在238 nm处检测的HPLC进行分析。在健康人脑脊液中HPETEs和HETEs区域的HPLC上未观察到峰。在SAH患者中,根据脑血管痉挛的发生识别出几个峰。其中一个峰通过HPLC和GC - MS鉴定为5 - HETE。在10例SAH患者中,通过测量HPLC上鉴定为5 - HETE的峰高对脑脊液中的5 - HETE进行半定量分析。在脑血管痉挛的发生与脑脊液中5 - HETE的出现之间发现密切相关性。本研究结果表明脂质过氧化参与SAH后慢性血管痉挛的发病机制。