出血性休克诱导的淋巴蛋白质组变化：损伤相关分子模式的出现。

Changes in lymph proteome induced by hemorrhagic shock: the appearance of damage-associated molecular patterns.

机构信息

Department of Surgery, Wayne State University, Detroit, MI, USA.

出版信息

J Trauma Acute Care Surg. 2012 Jul;73(1):41-50; discussion 51. doi: 10.1097/TA.0b013e31825e8b32.

DOI:10.1097/TA.0b013e31825e8b32

PMID:22743371

Abstract

BACKGROUND

Damage-associated molecular patterns (DAMPs) released from host tissue after trauma and hemorrhagic shock (HS) have been shown to activate polymorphonuclear cells (PMNs) and lead to acute lung injury and systemic inflammatory response syndrome. The avenue by which DAMPs reach the circulation is unclear; however post-HS lymph has been shown to contain biologically active mediators. We therefore studied the time course of DAMP detection in systemic lymph and the effect of isotonic versus hypertonic resuscitation on DAMPs production and PMN activation in vitro.

METHODS

A canine HS/hind-limb lymph cannulation model was used. Animals were bled to a mean arterial pressure of 40 mm Hg and were resuscitated with shed blood plus equivalent amounts of Na+as either lactated Ringer's solution or 7.5% hypertonic saline solution (HSS). Lymph samples were collected at baseline, end-shock, and at various times after resuscitation. DAMPs were isolated from lymph samples and detected by Western blot for high-mobility group box 1 and mitochondrial DNA. Priming of naive PMNs was indexed by mitogen-associated protein kinase phosphorylation. Human pulmonary microvascular endothelial cell monolayers were established and exposed to the various lymph samples. Endothelial intracellular adhesion molecule expression, apoptosis, and monolayer permeability were determined.

RESULTS

DAMPs were detected in lymph samples starting at the end of the shock period and peaking at 120 minutes after resuscitation. HSS resuscitation resulted in the highest levels of DAMPs detected in systemic lymph and plasma. PMN mitogen-associated protein kinase activation was noted during the resuscitation phase and peaked 120 minutes after resuscitation. Similar temporal changes in human pulmonary microvascular endothelial cell intracellular adhesion molecule expression and cellular injury were noted after shock with the greatest effect noted with the hypertonic saline resuscitation regimen.

CONCLUSION

Lymph represents an important avenue for the delivery of DAMPs into the systemic circulation after HS. HSS lead to a significant increase in DAMPs production in the model. This finding may account for the conflicting data regarding the salutary effects of HSS resuscitation noted in clinical versus experimental shock studies. ).

摘要

背景

创伤和失血性休克（HS）后宿主组织释放的损伤相关分子模式（DAMPs）已被证明可激活多形核细胞（PMN），导致急性肺损伤和全身炎症反应综合征。 DAMPs 进入循环的途径尚不清楚；然而，已经证明 HS 后淋巴液中含有具有生物活性的介质。因此，我们研究了 DAMPs 在全身淋巴中的检测时间过程，以及等渗与高渗复苏对体外 DAMPs 产生和 PMN 激活的影响。

方法

使用犬 HS/后肢淋巴插管模型。动物被放血至平均动脉压 40mmHg，并通过输注失血加等量的钠离子来复苏，分别使用乳酸林格氏液或 7.5%高渗盐水溶液（HSS）。在基线、休克结束时以及复苏后不同时间采集淋巴样本。通过 Western blot 从淋巴样本中分离 DAMPs，并检测高迁移率族 box1 和线粒体 DNA。通过有丝分裂原相关蛋白激酶磷酸化来标记幼稚 PMN 的启动。建立了人肺微血管内皮细胞单层，并暴露于各种淋巴样本。测定内皮细胞细胞间黏附分子表达、细胞凋亡和单层通透性。

结果

DAMPs 从休克期末开始在淋巴样本中检测到，并在复苏后 120 分钟达到峰值。HSS 复苏导致全身淋巴和血浆中检测到的 DAMPs 水平最高。PMN 有丝分裂原相关蛋白激酶的激活在复苏阶段观察到，并在复苏后 120 分钟达到峰值。在休克后，人肺微血管内皮细胞细胞间黏附分子表达和细胞损伤也出现了类似的时间变化，其中高渗盐水复苏方案的效果最大。