Department of Pharmacology, Physiology, and Therapeutics, University of North Dakota, 501 N. Columbia Road, Grand Forks, ND 58202-9037, USA.
Prostaglandins Leukot Essent Fatty Acids. 2013 Oct;89(5):313-8. doi: 10.1016/j.plefa.2013.09.005. Epub 2013 Sep 16.
Previously, we, and others, have demonstrated a rapid and significant post-mortem increase in brain prostanoid (PG) levels analyzed without microwave fixation, and this is not the result of PG trapping or destruction in microwave-irradiated brain tissue. In the present study, we demonstrate a dramatic increase in kidney eicosanoid levels when analyzed without microwave fixation which was mainly accounted for by the 142-, 81-, and 62-fold increase in medullary 6-ketoPGF1α, PGE2, and PGF2α, levels, respectively, while PGD2 and TXB2 levels were increased ~7-fold. Whole kidney and cortex PG were also significantly increased in non-microwaved tissue, but at lesser extent. Arachidonic acid and the lipoxygenase products hydroxyeicosatetraenoic acids (HETE) were also induced in whole kidney, cortex, and medulla 1.5- to 5.5-fold depending upon tissue and metabolite. Cyclooxygenase inhibition with indomethacin decreased PG mass in non-microwaved tissue to basal levels, however HETE and arachidonic acid were not decreased. These data demonstrate the critical importance of kidney tissue fixation to limiting artifacts during kidney eicosanoid analysis.
先前,我们和其他人已经证明,在未经微波固定的情况下,大脑前列腺素(PG)水平会迅速显著增加,而且这不是微波辐照脑组织中 PG 捕获或破坏的结果。在本研究中,我们发现未经微波固定分析时,肾脏花生四烯酸代谢产物水平会大幅增加,这主要归因于髓质中 6-酮-PGF1α、PGE2 和 PGF2α 水平分别增加了 142、81 和 62 倍,而 PGD2 和 TXB2 水平增加了约 7 倍。全肾和皮质 PG 在未经微波处理的组织中也显著增加,但程度较小。根据组织和代谢产物的不同,整个肾脏、皮质和髓质中的花生四烯酸和脂氧合酶产物羟二十碳四烯酸(HETE)也分别诱导了 1.5 到 5.5 倍。非微波处理组织中,环加氧酶抑制剂吲哚美辛将 PG 总量降低至基础水平,但 HETE 和花生四烯酸并未降低。这些数据表明,肾脏组织固定对于限制肾脏花生四烯酸代谢产物分析中的人为因素具有至关重要的意义。