Effects of Doxycycline on Renal Ischemia Reperfusion Injury Induced by Abdominal Compartment Syndrome

doi:10.1016/j.jss.2009.09.048

Journal of Surgical Research

Volume 167, Issue 1, 1 May 2011, Pages 113-120

https://doi.org/10.1016/j.jss.2009.09.048 Get rights and content

Background

The aim of this study was to determine the effects of doxycycline on renal ischemia reperfusion (I/R) injury in a rat model of abdominal compartment syndrome (ACS).

Materials and Methods

Forty-two Sprague-Dawley rats were divided into six groups. In the control group (group 1), kidney samples were collected with no manipulation; in the sham group (group 2) induction of ACS was followed by decompression. In groups 3 and 4, 1cc of saline was administered intraperitoneally (i.p.) during the induction of ACS, and the kidneys were removed 1 and 24h after decompression, respectively. In groups 5 and 6, doxycycline (10mg/kg i.p.) was injected during the induction of ACS, and similarly all tissue samples were removed 1 and 24h after decompression, respectively. MDA, IL-1β, IL-6, TNF-α, MMP-2, and TIMP-1 were studied, and the apoptotic cells were enumerated histopathologically, and apoptosis and bcl-2 expression were assessed immunohistochemically.

Results

Creatinine, MDA, IL-1β, and IL-6 levels were significantly higher in group 3, 1h after the reperfusion period compared with the control group, and the same parameters were significantly lower in the groups in which doxycycline was administered, 1hour after decompression. However, there remained no difference between groups at 24h, except IL-1β, which was decreased to even lower values. TNF-α and TIMP-1 levels were not statistically different in all groups. The MMP-2 level was significantly higher in group 4 by 24h, and there remained no difference between groups 1, 2, and 6. In group 6, there were not any apoptotic cells as were observed in the other groups. The number of apoptotic cells and the expression of bcl-2 was significantly less in the groups in which doxycycline was administered.

Conclusion

Doxycycline had protective effects on I/R injury by decreasing apoptosis via reducing the level of pro-inflammatory cytokines, increasing the level of TIMP-1, and inhibiting the activity of MMP-2.

Introduction

The pressure within the abdominal cavity is normally atmospheric or subatmospheric and intra-abdominal hypertension (IAH) affects each organ system separately at different levels of pressure [1]. Abdominal compartment syndrome (ACS) refers to organ dysfunction and ischemia resulting from IAH. However, increased intra-abdominal pressure reduces blood flow to intra-abdominal organs, and decompression may cause another serious problem, ischemia/reperfusion (I/R) injury 2, 3, 4.

It is well-known that reperfusion of ischemic tissue may promote the generation of various reactive oxygen metabolites, which have deleterious effects on cell membranes by mediating lipid peroxidation. MDA, which arises from the breakdown of lipid peroxyl radicals, is one of the indicators of oxidative stress. MDA is also important, in which it can cause further oxidative injury by oxidizing protein molecules [5].

Oxidants released during the reperfusion of ischemic tissue stimulate transcription factors involved in the expression of tumor necrosis factor-alpha (TNF-α). TNF-α stimulates the release of other inflammatory mediators, including interleukin-1, platelet-activating factor, oxygen radicals, and nitric oxide. It has been demonstrated that TNF-α is an important mediator in renal I/R injury and induces renal cell apoptosis, glomerular endothelial damage, and renal failure [5].

Matrix metalloproteinases (MMPs) are proteolytic enzymes that degrade the extracellular matrix and play important roles in inflammation, neoplastic invasion, and metastasis. The ubiquitous tissue inhibitor of metalloproteinases (TIMPs) can interfere with MMP proteolytic activation and enzymatic activity. MMPs are significantly activated in I/R injury in the lung, heart, and brain [6], as well as in kidneys 7, 8, 9. MMP expression by immune cells is greatly modulated by inflammatory mediators, such as TNF-α and IL-1β [10].

Apoptosis is a mechanism of cell death, which is characterized by the absence of cellular lysis and inflammation, and represents a crucial mechanism of I/R injury. Specifically, activation of proteases and phospholipases, alteration of intracellular calcium, ATP depletion, and cell damage by free oxygen radicals lead to cellular dysfunction [11]. The relationship between tissue I/R and apoptosis has been demonstrated in several organs, including the liver, brain, myocardium, and kidney. However, exactly how apoptosis is induced in different organs after I/R is still unclear, particularly in the kidney [12].

Doxycycline belongs to the family of tetracycline antibiotics and inhibits bacterial protein synthesis. Doxycycline binds directly to the active target site of MMPs and induces a degradation of the pro-MMP zymogen or provokes an inhibition of the transcription of MMP mRNAs [13]. Doxycycline may also reduce apoptosis associated with increased Bcl2 by inhibiting MMP-2 activity [14]. Doxycycline has anti-inflammatory effects by decreasing the expression of the genes encoding the pro-inflammatory cytokines, In addition to IL-1β and TNF-α [15], doxycycline may also up-regulate TIMP-1 expression [16]. All of these features of doxycycline may probably have protective roles in I/R injury.

In this study, we aimed to determine whether doxycycline has protective effects on the ACS, if there are any mechanisms by which it facilitates these effects, and any possible relationship with apoptosis.

Section snippets

Materials and Methods

Forty-two Sprague-Dawley male and female rats weighing 225–250 g were used in this study. All experimental protocols were approved by the Eskisehir Osmangazi University School of Medicine Animal Care and Use Committee. The rats were kept in individual cages and acclimated for at least 7 d before the experiments. All animals were kept in a temperature-controlled room with a 12-h light-dark cycle and free access to water and standard laboratory chow. Under ketamine anesthesia (100 mg/kg ketamine

Results

In group 3, creatinine level was significantly higher than groups 1 and 2 (P < 0.05). There was no difference between groups 5 and 1, 2 (P > 0.05). It was observed that creatinine level was decreased in group 4, and there remained no difference between groups 4 and 1, 2 (P > 0.05). There was no difference between groups 3, 4, 5, and 6 (Table 1).

TNF-α, IL-1β, and IL-6 levels were significantly higher in group 3 1 h after the reperfusion period following ischemia compared with the control group (TNF-α, P

Discussion

The protective effects of doxycycline and its derivative, minocycline, against I/R injury and oxidative stress have been previously studied 17, 18, 19, 20. However, for the first time, we observed that doxycycline has protective effects on I/R injuries of kidneys induced by ACS, and we demonstrated the possible physiologic ways by which this benefit occurs, including down-regulation of pro-inflammatory cytokines and MMPs, up-regulation of bcl-2, and inhibition of apoptosis.

According to the

Acknowledgments

This project was supported by the Eskisehir Osmangazi University Scientific Research Projects Commission.

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Shock/Sepsis/Trauma/Critical CareEffects of Doxycycline on Renal Ischemia Reperfusion Injury Induced by Abdominal Compartment Syndrome

Background

Materials and Methods

Results

Conclusion

Introduction

Section snippets

Materials and Methods

Results

Discussion

Acknowledgments

Peptides

Surgery

Transplant Proc

Free Radic Biol Med

Biochem Pharmacol

Biochem Biophys Res Commun

Neurochem Int

Biochem Pharmacol

J Biol Chem

Kidney Int

Abdominal compartment syndrome

J Trauma

Systemic inflammatory response secondary to abdominal compartment syndrome: Stage for multiple organ failure

J Trauma

Gut ischemia, oxidative stress, and bacterial translocation in elevated abdominal pressure in rats

World J Surg

Effect of leptin on renal ischemia-reperfusion damage in rats. J Physiol

Biochem

Shock/Sepsis/Trauma/Critical Care
Effects of Doxycycline on Renal Ischemia Reperfusion Injury Induced by Abdominal Compartment Syndrome