Elrod K, Okamoto H, Greenbaum L M, Buccafusco J J
Neurochem Res. 1986 Oct;11(10):1463-71. doi: 10.1007/BF00966225.
Focused microwave irradiation was employed to stabilize endogenous whole rat brain bradykinin levels prior to a simple extraction procedure. Skull microwave exposure (2450 MHz, 3.8 kW., 2.45 sec) resulted in inactivation to less than 5% of control of whole brain kallikrein and kininase activity. Using this adequate exposure duration whole rat brain kinin levels as measured by a sensitive radioimmunoassay were approximately 0.6 pmol/g (wet weight). Further purification of irradiated brain extracts using HPLC revealed that immunoreactive kinin eluted as a single peak that co-chromatographed with authentic bradykinin. Microwave fixation duration of 1.25 sec yielded greatly increased levels of immunoreactive kinin which following HPLC purification eluted in two peaks, corresponding to authentic bradykinin and T-kinin, respectively. The tissue injury resulting from incomplete microwave fixation resulted in the release of kinins. This excess immunoreactive kinin may be derived from cerebral blood, since the predominant form of kinin-generating protein in plasma is T-kininogen.
在进行简单提取程序之前，采用聚焦微波辐射来稳定内源性全大鼠脑缓激肽水平。颅骨微波暴露（2450 MHz，3.8 kW，2.45秒）导致全脑激肽释放酶和激肽酶活性失活至对照的5%以下。使用这个适当的暴露持续时间，通过灵敏的放射免疫测定法测得的全大鼠脑激肽水平约为0.6 pmol/g（湿重）。使用高效液相色谱法（HPLC）对辐照后的脑提取物进行进一步纯化，结果显示免疫反应性激肽以单一峰形式洗脱，该峰与 authentic bradykinin 共色谱。1.25秒的微波固定持续时间使免疫反应性激肽水平大幅增加，经HPLC纯化后，分别在两个峰中洗脱，对应于 authentic bradykinin 和T-激肽。不完全微波固定导致的组织损伤导致激肽释放。这种过量的免疫反应性激肽可能源自脑血，因为血浆中激肽生成蛋白的主要形式是T-激肽原。
