Thongboonkerd Visith, Chiangjong Wararat, Mares Jan, Moravec Jiri, Tuma Zdenek, Karvunidis Thomas, Sinchaikul Supachok, Chen Shui-Tein, Opatrný Karel, Matejovic Martin
Medical Proteomics Unit, Office for Research and Development, Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand.
Clin Sci (Lond). 2009 May;116(9):721-30. doi: 10.1042/CS20080478.
Sepsis is a systemic response to infection commonly found in critically ill patients and is associated with multi-organ failure and high mortality rate. Its pathophysiology and molecular mechanisms are complicated and remain poorly understood. In the present study, we performed a proteomics investigation to characterize early host responses to sepsis as determined by an altered plasma proteome in a porcine model of peritonitis-induced sepsis, which simulated several clinical characteristics of human sepsis syndrome. Haemodynamics, oxygen exchange, inflammatory responses, oxidative and nitrosative stress, and other laboratory parameters were closely monitored. Plasma samples were obtained from seven pigs before and 12 h after the induction of sepsis, and plasma proteins were resolved with two-dimensional gel electrophoresis (n=7 gels/group; before being compared with during sepsis). The resolved proteins were stained with the SYPRO Ruby fluorescence dye and subjected to quantitative and comparative analyses. From approx. 1500 protein spots visualized in each gel, levels of 36 protein spots were significantly altered in the plasma of animals with sepsis (sepsis/basal ratios or degrees of change ranged from 0.07 to 21.24). Q-TOF (quadrupole-time-of-flight) MS and MS/MS (tandem MS) identified 30 protein forms representing 22 unique proteins whose plasma levels were increased, whereas six forms of five unique proteins were significantly decreased during sepsis. The proteomic results could be related to the clinical features of this animal model, as most of these altered proteins have important roles in inflammatory responses and some of them play roles in oxidative and nitrosative stress. In conclusion, these findings may lead to a better understanding of the pathophysiology and molecular mechanisms underlying the sepsis syndrome.
脓毒症是危重症患者中常见的一种对感染的全身性反应，与多器官功能衰竭和高死亡率相关。其病理生理学和分子机制复杂，目前仍知之甚少。在本研究中，我们进行了一项蛋白质组学研究，以表征在腹膜炎诱导的脓毒症猪模型中，由血浆蛋白质组改变所确定的宿主对脓毒症的早期反应，该模型模拟了人类脓毒症综合征的若干临床特征。对血流动力学、氧交换、炎症反应、氧化和亚硝化应激以及其他实验室参数进行了密切监测。在脓毒症诱导前和诱导后12小时从7头猪获取血浆样本，并用二维凝胶电泳分离血浆蛋白（每组n = 7块凝胶；在脓毒症发生前与发生期间进行比较）。分离出的蛋白质用SYPRO Ruby荧光染料染色，并进行定量和比较分析。在每块凝胶中大约可看到1500个蛋白点，脓毒症动物血浆中36个蛋白点的水平发生了显著变化（脓毒症/基础比值或变化程度范围为0.07至21.24）。Q-TOF（四极杆-飞行时间）质谱和串联质谱鉴定出30种蛋白形式，代表22种独特的蛋白质，其血浆水平在脓毒症期间升高，而5种独特蛋白质的6种形式在脓毒症期间显著降低。蛋白质组学结果可能与该动物模型的临床特征相关，因为这些改变的蛋白质大多数在炎症反应中具有重要作用，其中一些在氧化和亚硝化应激中发挥作用。总之，这些发现可能有助于更好地理解脓毒症综合征的病理生理学和分子机制。
