Elsevier

Resuscitation

Volume 82, Issue 6, June 2011, Pages 760-766
Resuscitation

Experimental paper
Limb ischemic preconditioning mitigates lung injury induced by haemorrhagic shock/resuscitation in rats

https://doi.org/10.1016/j.resuscitation.2011.02.010Get rights and content

Abstract

Aim of the study

Haemorrhagic shock and subsequent resuscitation induce acute lung injury. We elucidated whether bilateral lower limb ischemic pre-conditioning (IP) could mitigate lung injury in haemorrhagic shock/resuscitation rats. The role of heme oxygenase-1 (HO-1) was also elucidated.

Method

Adult male rats were randomized to receive haemorrhagic shock/resuscitation (HS), HS plus IP, or HS plus IP plus the HO-1 inhibitor tin protoporphyrin (SnPP) (n = 12 in each group). Sham groups were employed simultaneously. For pre-conditioning, 3 cycles of limb IP (10 min ischemia followed by 10 min reperfusion) were performed immediately before haemorrhagic shock. Haemorrhagic shock (mean arterial pressure: 40–45 mmHg) was induced by blood drawing and maintained for 120 min. SnPP was injected 5 min before resuscitation. Shed blood/saline mixtures were re-infused to achieve resuscitation. After monitoring for another 8 h, rats were sacrificed. Arterial blood gas and alveolar–arterial oxygen difference (lung function index), histology, polymorphonuclear leukocytes/alveoli ratio (leukocyte infiltration index), wet/dry weight ratio (water content index), inflammatory molecules (e.g., chemokine, cytokine, prostaglandin E2), and malondialdehyde (lipid peroxidation index) assays were preformed.

Results

Haemorrhagic shock/resuscitation induced significant lung function alterations and significant increases in leukocyte infiltration, water content, inflammation, and lipid peroxidation in lungs. Histological analysis confirmed that haemorrhagic shock/resuscitation caused marked lung injury. Limb IP significantly mitigated the adverse effects of haemorrhagic shock/resuscitation. Moreover, the protective effects of limb IP were reversed by SnPP.

Conclusions

Limb IP mitigates lung injury in haemorrhagic shock/resuscitation rats. The mechanisms may involve HO-1.

Introduction

Haemorrhagic shock followed by resuscitation (i.e., haemorrhagic shock/resuscitation) is a status of global ischemia and reperfusion (I/R) that readily causes lung injury.1, 2 Oxidative stress mediates the development of lung injury induced by haemorrhagic shock/resuscitation by activating neutrophils and eliciting the inflammation response in lungs.1,2 Oxidants, infiltrated leukocytes, and inflammatory molecules then work together to injure lung endothelial cell and result in microvascular integrity loss, leading to impaired gas exchange and lung dysfunction.3

Limb ischemic pre-conditioning (IP), a process of transient I/R, is an adaptation response that can protect organs from injuries imposed by prolonged I/R.4 Previous data demonstrated that limb IP could ameliorate liver dysfunction induced by liver I/R.5 Limb IP also mitigated lung injury induced by prolonged lower limb I/R.6 Myocardial infarct size after coronary artery ligation and reperfusion was also decreased by limb IP.7

The effects of limb IP on modulating lung injury induced by haemorrhagic shock/resuscitation, however, has yet to be studied. We thus conducted this study with the hypothesis that limb IP attenuates lung injury in haemorrhagic shock/resuscitation rats. We also examined the question of whether heme oxygenase-1 (HO-1), a stress response protein with potent anti-oxidation and anti-inflammation capacity,8 participates in the protective effects of limb IP on lung injury induced by haemorrhagic shock/resuscitation because previous data9 indicated that HO-1 might involve in the effects of limb IP on attenuating lung injury induced by lower limb IR.

Section snippets

Methods

This study was approved by the Animal Use and Care Committee, Buddhist Tzu Chi General Hospital, Taipei Branch. Care and handling of rats were in accordance with the National Institute of Health guidelines. A total of 72 adult male Sprague–Dawley rats (age: 50–56 days old; body weight: 200–250 g; BioLASCO Taiwan Co., Ltd., Taipei, Taiwan) were used.

Results

With the exception of HO-1, data of the Sham–IP and Sham–IP–SnPP groups were not significantly different from those of the sham group. We thus decided to omit most of the data from the Sham–IP and Sham–IP–SnPP groups and only report the HO-1 data in the following session.

Discussion

This study confirmed that limb IP mitigates lung injury in haemorrhagic shock/resuscitation rats. Findings from this study support the concept that limb IP could serve as an effective adjunct to therapies against injuries in situations where haemorrhagic shock/resuscitation is predicted, e.g., scheduled major surgeries.

Our data that reactive nitrogen species (e.g., NO) and malondialdehyde as well as inflammatory molecules (e.g., MIP-2, IL-6, and PGE2) in lungs were increased in the HS group

Conclusions

Limb IP mitigates acute lung injury induced by haemorrhagic shock/resuscitation. The mechanisms are mediated, at least in part, by HO-1.

Conflicts of interest statement

We stated that there are no potential conflicts of interest, including employment, consultancies, stock ownership, honoraria, paid expert testimony, patent applications/registrations, and grants or other funding, for each author to be disclosed.

Acknowledgments

This work was performed mainly in Buddhist Tzu Chi General Hospital, Taipei Branch and supported by grants from National Science Council, Taiwan (NSC 98-2314-B-303-012-MY3) awarded to Dr. Huang and Mackay Medicine, Nursing and Management College (MKC99R19) awarded to Dr. Jan.

Role of the funding source: We state that the study sponsors play no roles in study design; in the collection, analysis, and interpretation of data; in the writing of the report; and in the decision to submit the paper for

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    A Spanish translated version of the abstract of this article appears as Appendix in the final online version at doi:10.1016/j.resuscitation.2011.02.010.

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