Hinkelbein J, Kalenka A, Feldmann R E
Klinik für Anästhesiologie und Operative Intensivmedizin, Universitätsmedizin Mannheim (UMM), Theodor-Kutzer-Ufer 1-3, 68167 Mannheim, Deutschland.
Anaesthesist. 2009 Feb;58(2):134-43. doi: 10.1007/s00101-008-1488-6.
Sepsis still has a high mortality in critically-ill patients. Here, analysis of early alterations in cerebral proteome may lead to a better understanding of the molecular basis of cerebral dysfunction. The aim of the present study was therefore to analyze cerebral protein dynamics during emerging sepsis in an established rat model.
To induce sepsis, an established coecal ligature and double puncture (coecal ligature and puncture, CLP) model was used on Wistar rats. After 12 hours, surviving rats (sepsis: n=6 and sham: n=6) were decapitated and their brains prepared for gel electrophoresis (2DE) and subsequent mass spectrometry. Biological function of differentially regulated proteins (t-test, p<0.01) was then analyzed using bioinformatic network analysis (ingenuity pathways analysis, IPA).
Mortality was 40 % in the sepsis-group and no rat of the sham-group died. Altogether, nine significantly regulated proteins were identified (4 up-regulated, 5 down-regulated). IPA then detected eight network proteins and interpreted them in the context of established protein alterations for sepsis.
The combination of proteomics and IPA could identify proteins in rat brain, whose expression was significantly regulated during sepsis. The methodological approach applied in the present study may facilitate the quest for novel sepsis-induced protein alterations in the future.
脓毒症在重症患者中死亡率仍然很高。在此,对脑蛋白质组早期变化的分析可能有助于更好地理解脑功能障碍的分子基础。因此,本研究的目的是在一个成熟的大鼠模型中分析脓毒症发生过程中的脑蛋白质动态变化。
为诱导脓毒症,在Wistar大鼠上采用成熟的盲肠结扎和双穿刺(盲肠结扎和穿刺,CLP)模型。12小时后,将存活的大鼠(脓毒症组:n = 6;假手术组:n = 6)断头,并将其大脑用于凝胶电泳(2DE)及后续质谱分析。然后使用生物信息网络分析( Ingenuity通路分析,IPA)分析差异调节蛋白(t检验,p < 0.01)的生物学功能。
脓毒症组死亡率为40%,假手术组无大鼠死亡。共鉴定出9种显著调节的蛋白(4种上调,5种下调)。IPA随后检测到8种网络蛋白,并在脓毒症既定的蛋白变化背景下对其进行了解释。
蛋白质组学和IPA的结合能够鉴定大鼠脑中在脓毒症期间表达受到显著调节的蛋白。本研究中应用的方法可能有助于未来探索新型脓毒症诱导的蛋白变化。