Weykamp Michael, Nikolian Vahagn C, Dennahy Isabel S, Higgins Gerald A, Georgoff Patrick E, Remmer Henriette, Ghandour Mohamed H, Alam Hasan B
Department of Surgery, University of Michigan Medical School, Ann Arbor, Michigan.
Department of Computational Medicine and Bioinformatics, University of Michigan Medical School, Ann Arbor, Michigan.
J Surg Res. 2018 Aug;228:84-92. doi: 10.1016/j.jss.2018.02.046. Epub 2018 Apr 25.
Histone deacetylase inhibitors such as valproic acid (VPA) improve survival in lethal models of hemorrhagic shock and polytrauma. Although VPA is known to modulate transcription, its ability to reduce mortality within minutes of administration suggests involvement of a rapid, posttranslational mechanism. We hypothesized that VPA treatment would cause proteomic changes within minutes of treatment including quantitative and/or posttranslational differences in structural and/or effector proteins.
We used a porcine model of traumatic brain injury (computer-controlled cortical impact, 12 mm depth) and hemorrhagic shock (40% hemorrhage). Animals were kept in shock for 2 h and randomized to two groups (n = 3): normal saline (volume = 3:1 hemorrhage volume) or normal saline + VPA (150 mg/kg, single dose). Peripheral blood mononuclear cells were collected at baseline, postshock, and postresuscitation. Intracellular protein profiles were assessed using 1 dimensional gel electrophoresis, liquid chromatography, mass spectrometry, and analyzed with Ingenuity Pathway Analysis software.
Animals treated with VPA demonstrated significant proteomic changes. Quantitative differences were found in over 200 proteins including effector, regulatory, and structural proteins in critical cell signaling pathways. Posttranslational modification analysis demonstrated differential VPA-induced acetylation of lysine residues in histone and nonhistone proteins. Pathway analysis correlated these changes with significant increases in numerous prosurvival and cytoskeletal intracellular pathways, including Rho GTPase signaling (P = 1.66E-11), integrin signaling (P = 4.19E-21), and a decrease in Rho guanosine nucleotide dissociation inhibitor signaling (P = 4.83E-12).
In a porcine model of severe injuries, a single dose of VPA is associated with protective changes in the proteome that are measurable within minutes of treatment.
组蛋白脱乙酰酶抑制剂如丙戊酸（VPA）可提高失血性休克和多发伤致死模型的生存率。尽管已知VPA可调节转录，但其在给药数分钟内降低死亡率的能力提示涉及一种快速的翻译后机制。我们推测VPA治疗会在治疗数分钟内引起蛋白质组学变化，包括结构和/或效应蛋白的定量和/或翻译后差异。
我们使用了猪创伤性脑损伤模型（计算机控制的皮质撞击，深度12毫米）和失血性休克模型（40%出血）。动物在休克状态下维持2小时，随机分为两组（n = 3）：生理盐水组（容量 = 出血体积的3倍）或生理盐水 + VPA组（150毫克/千克，单次剂量）。在基线、休克后和复苏后采集外周血单个核细胞。使用一维凝胶电泳、液相色谱、质谱评估细胞内蛋白质谱，并使用Ingenuity Pathway Analysis软件进行分析。
接受VPA治疗的动物表现出显著的蛋白质组学变化。在关键细胞信号通路中的200多种蛋白质中发现了定量差异，包括效应蛋白、调节蛋白和结构蛋白。翻译后修饰分析表明，VPA诱导组蛋白和非组蛋白中赖氨酸残基的乙酰化存在差异。通路分析将这些变化与许多促生存和细胞骨架细胞内通路的显著增加相关联，包括Rho GTPase信号通路（P = 1.66E - 11）、整合素信号通路（P = 4.19E - 21），以及Rho鸟苷核苷酸解离抑制剂信号通路的减少（P = 4.83E - 12）。
在严重损伤的猪模型中，单次剂量的VPA与治疗数分钟内可测量的蛋白质组保护性变化相关。
