Skip to main content

Doxycycline reduces liver and kidney injuries in a rat hemorrhagic shock model

Abstract

Background

Hemorrhagic shock (HS), which causes insufficient tissue perfusion, can result in multiple organ failure (MOF) and death. This study aimed to evaluate whether doxycycline (DOX) protects cardiovascular, kidney, and liver tissue from damage in a rat model of HS. Immediately before the resuscitation, DOX (10 mg/kg; i.v.) was administered, and its protective effects were assessed 24 h later. Mean arterial pressure, renal blood flow, heart rate, vasoactive drug response, and blood markers such as urea, creatinine, AST, ALT, CPK, CPR, and NOx levels were determined.

Results

We showed that DOX has a significant effect on renal blood flow and on urea, creatinine, AST, ALT, CPK, and NOx. Morphologically, DOX reduced the inflammatory process in the liver tissue.

Conclusions

We conclude that DOX protects the liver and kidney against injury and dysfunction in a HS model and could be a strategy to reduce organ damage associated with ischemia-and-reperfusion injury.

Background

Multiple organ failure can be the result of trauma injuries, which is one of the main causes of mortality among young people and adults, resulting in roughly 5 million deaths each year [1], with half of these deaths caused by severe hemorrhage and hemorrhagic shock (HS). During HS, the reduction of blood volume causes insufficient tissue perfusion, resulting in an imbalance between oxygen consumption and supply, which can lead to multiple organ failure (MOF) [2].

Blood levels of several molecules that are used to identify injuries and dysfunctions, such as urea, creatinine, aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatine phosphokinase (CPK), C-reactive protein (CRP), and nitrite and nitrate (NOx), may be monitored to verify if any organ injuries have taken place [3]. Additionally, other molecules, such as matrix metalloproteinases, are often produced by inflammatory cells and have an important role in the process of organ injury [4].

Unfortunately, there is no specific treatment to prevent or minimize MOF associated with HS [5]. As a result, researchers are interested in investigating the effects of approved drugs, at least in experimental models of HS [6,7,8,9].

In this context, doxycycline (DOX), a member of the tetracycline family of antibiotics used to treat various bacterial infections, has been exhibiting potential clinical benefits. It also exhibits pleiotropic effects, including anti-inflammatory and antioxidant properties [10, 11]; suppresses nitric oxide (NO) overproduction [12, 13], and matrix metalloproteinases (MMPs) activity. Both molecules (NO and MMPs) are strongly related to poor outcomes in severe situations including sepsis and HS [3, 14, 15].

It's interesting to note that DOX effects have been studied in both human and animal models for a variety of diseases like cardiac disease, arthritis, periodontitis, and chronic wounds [11, 14, 16, 17] that are characterized by increased MMP activity and inflammation.

Despite DOX has been tested in animal models of sepsis and even HS-related MOF [3, 18], its effects in MOF associated with a recovery model of HS, particularly in the morphology of important organs such as the liver and kidney are unknown.

Therefore, using our stablished protocol to create an experimental HS condition [7, 8, 19, 20], the aim of this study was to evaluate the protective effect of DOX when administered at the resuscitation in a rat model of HS.

Methods

Animals

Male Wistar rats (200–250 g) were housed in a temperature- and light-controlled room (23 ± 2 °C; 12 h light/dark cycle) with free access to water and food. Five rats per cage were kept in 45 × 34 × 16 cm plastic cages. All experiments were performed between 9:00 and 16:00 h. Animal procedures are in accordance with the National Institutes of Health Animal Care Guidelines and the Guide of the Brazilian National Council of Animal Experimentation. Animal studies are reported in compliance with the ARRIVE guidelines. The procedures were approved by the Universidade Estadual de Ponta Grossa Ethics Committee (protocol number 048/2015).

Hemorrhagic shock procedure and experimental groups

The HS model was performed as previously described with minor modifications [7, 8]. Rats were anesthetized with sodium thiopentone (100 mg/kg i.p.). Left femoral artery (for blood withdrawn and blood pressure measurement) and vein (for resuscitation and drug delivery) were cannulated (polythene tubing; PP25; Smiths Medical International Ltd, Kent, UK). Blood was withdrawn from the artery (at a rate of 1 mL/min) and collected with heparin (2 IU per mL of blood; Hepamax- Blausiegel, Cotia, SP, Brazil) until the mean arterial pressure (MAP) reached 40 ± 2 mmHg, which was maintained for 90 min either by further withdrawal of blood during the compensation phase or administration of the shed blood during the decompensation phase. Blood pressure was recorded with a catheter pressure transducer coupled to a Powerlab 4/30 (ADInstruments Pty Ltd.; Castle Hill, New South Wales, Australia) running the proprietary software LabChart 8®. The shed blood was kept between 6 and 10 °C. Immediately before the resuscitation with blood, HS animals were randomly divided to receive vehicle (saline; 1 mL/kg; i.v.) or doxycycline (DOX; 10 mg/kg/mL; i.v.) in bolus through the femoral vein. Then, resuscitation was performed with the shed blood (warmed to 22 ± 2 °C) over a period of 5 min followed by 1.5 mL/kg of Ringer’s lactate (Eurofarma-Ribeirão Preto, SP, Brazil). Twenty minutes after the end of resuscitation, cannulas were removed, vessels were ligated, and the skin was sutured. All animals were allowed to recover from anaesthesia in a warm cage. Sham-rats were used as control and underwent identical surgical procedures, but without haemorrhage or resuscitation, and were also randomly treated with DOX or saline. Twenty-four hours after resuscitation, recovery animals were anesthetized once more for assessment of all parameters described below. One group of animals underwent cardiovascular analyses (mean arterial pressure, heart rate, and response to vasoactive drugs). A second group of animals underwent renal blood flow determination, as well as the extraction of liver and blood samples as detailed below.

Cardiovascular parameters

In the first set of experiments, cardiovascular parameters were determined in sham and HS rats. Twenty-four hours after HS or sham surgery, animals were anesthetized with sodium thiopentone at dose 100 mg/kg i.p. (Cristalia—Itapira, SP, Brazil), and the jugular vein (for drug administration) and right carotid artery (for blood pressure measurement) were cannulated. MAP and heart rate (HR) data were recorded with a catheter pressure transducer coupled to a Powerlab 4/30 (ADInstruments Pty Ltd.; Castle Hill, New South Wales, Australia). Baseline MAP and HR were determined through the software LabChart 8®. In addition, rats were injected intravenously with acetylcholine (ACh) 0.1, 1, 10 nmol/kg, sodium nitroprusside (SNP) 3, 10, 30 nmol/kg), phenylephrine (Phe) 3, 10, 30 nmol/kg) and angiotensin II (Ang II) 3, 10, 30 pmol/kg (Sigma-Aldrich Company Ltd, Poole, Dorset, UK). The change in MAP (area under the curve from baseline—arbitrary units; AU) was calculated through the LabChart 8® software. Although a dose–response curve has been performed, for the sake of clarity, only the intermediate dose of each drug is shown in the graphs. At the end of the experiments, animals received an overdose of anaesthetic.

Renal blood flow (RBF)

In a separated group of animals, animals were anesthetized with sodium thiopentone at dose 100 mg/kg i.p. (Cristalia—Itapira, SP, Brazil). An abdominal incision was done to assess the left kidney, and a laser probe (model VP3) connected to a laser Doppler blood flow monitor (moorVMSLDF2, Moor Instruments, England) was placed directly on the kidney surface. Baseline RBF was determined through the software LabChart 8®. Immediately after RBF measurement, blood from all animals was obtained through cardiac puncture (exsanguination), and the livers were harvested. Blood was centrifuged (3000 g, 10 min) and the plasma was obtained and kept at −80 °C for the analyses described below.

Markers of organ injury and dysfunction

Plasma was obtained for urea, creatinine, aspartate aminotransferase (AST), alanine aminotransferase (ALT) and creatine phosphokinase (CPK) measurement through commercially available clinical assay kits (LabTest Diagnóstica S.A., Lagoa Santa, MG, Brazil). Plasma C-reactive protein (CRP) was quantified through a highly sensitive rat enzyme-linked immunosorbent assay kit (Immunology Consultants Laboratory Inc.; Newberg, OR, USA). The results were expressed as mg/mL. Plasma nitrite and nitrate (NOx) were zinc sulfate-deproteinized and subjected to nitrate conversion to nitrite using Escherichia coli nitrate reductase for 3 h at 37 °C. Samples were centrifuged for bacteria removal, and 100 µL of each sample was mixed with Griess reagent (1% sulphanilamide in 10% phosphoric acid / 0.1% naphtylethylenediamine) in a 96-well plate and read at 540 nm in a plate reader. Standard curves of nitrite and nitrate were run simultaneously, and values were expressed as µM.

Histology

Twenty-four hours after surgery, liver tissues were harvested, fixed (paraformaldehyde 4%) and embedded in paraffin. Sections (5 µm) were cut and stained with hematoxylin–eosin (HE) and examined under light microscopy. To quantify inflammation in liver tissue, 10 random fields (1000 ×) were evaluated, and polymorphonuclear (PMN) cells were counted.

Drugs

Unless otherwise stated, all compounds were from Sigma-Aldrich Company Ltd (Poole, Dorset, UK). Ringer’s lactate was from Eurofarma-Ribeirão Preto, SP, Brazil); sodium thiopentone (Thiopentax) from Cristalia (Itapira, SP, Brazil); heparin (Hepamax) from Blausiegel (Cotia, SP, Brazil).

Statistical analysis

The GPower 3.1.1 software was used for sample calculation, which was based on the SD and the magnitude of difference between the groups obtained in the analysis of MAP (mmHg) from our previous studies [8]. Considering 4 experimental groups, alpha = 0.05, and a power of 80%, 7 animals were required for statistical significance. As a certain mortality rate was expected, 10 animals were assigned per group, and the final number (n) in each group is indicated in the figures. When necessary, the values were transformed into logarithms to achieve normality and homogeneity of variances, verified by the Shapiro–Wilk and Bartlett tests, respectively. The results are presented as dot plot showing the mean ± SD. Statistical significance (p < 0.05) was analysed by ANOVA followed by Tukey’s post hoc test as indicated in the legends of figures. Pearson correlations were computed to examine the strength of associations in normally distributed samples (Fig. 2B–E). The statistical analyses were performed using GraphPad Prism® version 9.0 software (California, USA).

Results

Doxycycline attenuated renal dysfunction and liver injury

When compared with sham rats, HS animals exhibited a decrease in RBF (Fig. 1A), along with elevated levels of plasma urea (Fig. 1B) and creatinine (Fig. 1C), suggesting renal dysfunction. HS rats also developed liver injury characterized by increased levels of AST (Fig. 1D) and ALT (Fig. 1E), and muscular injury, as observed by increase of CPK (Fig. 1F). The treatment of HS rats with DOX prevented the decrease of RBF and reduced the plasma markers of organ damage.

Fig. 1
figure 1

Measurement for renal blood flow (A); plasma urea (B); plasma creatinine (C); plasma aspartate aminotransferase (AST) (D); plasma alanine aminotransferase (ALT) (E); and CPK, plasma creatine phosphokinase (F) in control groups (Sham) and HS treated with vehicle or doxycycline. *P < 0.05 vs. sham + vehicle, and #P < 0.05 vs. HS + vehicle

Doxycycline reduced significantly NOx production in HS rats

Rats in the HS group had significantly increased NOx levels compared to rats in the Sham control group (Fig. 2A). The administration of DOX in HS rats decreased NOx levels when compared to HS vehicle group. Additionally, the NOx levels showed a positive correlation (p < 0.05) with urea and creatinine (Fig. 2B, C), but not with AST, ALT (Fig. 2D, E) or CPK (data not shown).

Fig. 2
figure 2

Measurements for NOx production (A) and its correlations with urea (B), creatinine (C), plasma aspartate aminotransferase (AST) (D); plasma alanine aminotransferase (ALT) in control groups (Sham) and HS treated with doxycycline. *P < 0.05 vs. sham + vehicle, and #P < 0.05 vs. HS + vehicle

Doxycycline had no effect on mean arterial pressure (MAP), heart rate (HR) in HS rats, or C-reactive protein (CRP) levels

Twenty-four hours after HS, the baseline values of MAP (Fig. 3A) and HR (Fig. 3B) were similar in all experimental groups. However, the HS group showed a significant hypo-responsiveness to Ang II (Fig. 3C), and DOX therapy for HS rats failed to restore this response. Additionally, there was no difference among the groups for Phe, ACh, SNP responses, and for CRP levels (Fig. 3D–G, respectively).

Fig. 3
figure 3

Measurement for mean arterial pressure (mmHg) (A), heart rate (bpm) (B), response to angiotensin II (Ang II; 10 pmol/kg) (C), response to phenylephrine (Phe; 10 nmol/kg) (D), response to acetylcholine (ACh; 1 nmol/kg) (E), response to sodium nitroprusside (SNP; 10 nmol/kg) (F) and for C-reactive protein (G) in control groups (Sham) and HS treated with doxycycline. *P < 0.05 vs. sham + vehicle, and #P < 0.05 vs. HS + vehicle

Doxycycline reduced the inflammation process in liver of HS rats

Comparing the Sham group (Fig. 4A) to the HS group (Fig. 4C), the inflammatory cells into the liver were significantly increased in the HS group. However, after DOX treatment, the number of inflammatory cells was significantly reduced in the HS group (Fig. 4D, E).

Fig. 4
figure 4

Representative image of liver stained with hematoxilin/eosin for control groups Sham (A, B) and HS treated with doxycycline (B, D) and results for number of PMN in all groups (E). Arrow indicate PMN in sinusoidal vessels. *P < 0.05 vs. sham + vehicle animals. #P < 0.05 vs. HS + vehicle

Discussion

The primary goal of this investigation was to assess the protective effect of DOX when given before resuscitation in a rat model of HS. In order to do that, blood markers were used to infer kidney, liver and cardiovascular dysfunctions as well as the degree of morphological damage caused by inflammation in the liver.

As expected, the HS condition caused a number of alterations in all parameters assessed, such as a decrease in renal blood flow and elevated plasma levels of urea and creatinine, which may indicate kidney dysfunction. Additionally, the elevated levels of AST, ALT, and CPK indicate that HS model was associated with liver and muscle injury.

HS pathogenesis is invariably associated with hypoxia and ischemia–reperfusion damage. Reactive oxygen species (ROS) generation rises as blood flows through an ischemic tissue, which triggers the release of more inflammatory mediators. Together, these mediators contribute to organ and cell damage, which results in multiple organ failure and dysfunction [21].

In our study, the model of HS employed was found to reproduce the organ dysfunction observed in patients that suffered severe hemorrhage. Importantly, DOX was administered as a treatment immediately preceding the resuscitation with blood. The timing of the treatment was aimed at enhancing the clinical relevance of our study. When we examined how DOX protects the advance of some signs of organ damage, we found that DOX treatment in HS rats reduced liver injury, renal dysfunction, and CPK, an indication of muscle damage. Also, the improvement of kidney dysfunction in HS rats was confirmed with the return of RBF to sham values. Our results corroborates and extends a previous study that evaluated the effect of DOX on an acute model of HS and showed that DOX reduces liver damage and kidney dysfunction [22]. However, the mouse terminal model of HS used by the authors had a 6-h outcome period, in contrast to our investigation, which focused on the effects of DOX 24 h after the injury (a recovery model). In this previous study, they also evaluated the death rate over a week and discovered that DOX provided persistent protection.

Neutrophils produce cytokines, ROS, and inflammatory response enzymes such as myeloperoxidase, all of which have been connected to organ damage and dysfunction. The protective effect of DOX on liver injury indicated by plasma markers such as AST and ALT may have been generated, at least in part, by the decrease in the number of inflammatory cells seen in the liver of HS rats treated with DOX. Previous studies have reported a protective role of DOX against ischemia and reperfusion injury in hepatocytes. They show that the cytoprotection was mediated by the inhibition of the mitochondrial calcium uniporter rather than MMP inhibition [23]. However, if in our model this was the mechanism for the attenuation of liver inflammatory process, it needs to be determined.

Treatment with DOX has been associated with an improvement of cardiovascular outcome due to its antioxidant effect (reviewed in [11]). As HS is associated with an increase of ROS, antioxidant therapies could be beneficial to prevent cardiovascular dysfunction. The fact that MAP and HR were similar across all groups suggests that DOX therapy had no negative impact on cardiovascular parameters evaluated in our study. However, contrary to our expectations, no improvement on the response to Ang II was observed in HS rats.

Interestingly, the metabolites of nitric oxide (NOx) were reduced in plasma of HS rats treated with DOX. In pathological conditions, NO is mainly derived from nitric oxide synthase (NOS-2), which has been previously shown to be an important mediator of organ injury in HS [7, 8] and also in other conditions such as sepsis [24,25,26,27]. The upregulation of NOS-2 expression is observed in the initial hours following both septic shock [27] and HS [28]. In both conditions, the nitric oxide (NO) derived from NOS-2 has been documented to play a role in cardiovascular dysfunction and uncontrolled inflammation. This suggests noteworthy parallels between the pathophysiological mechanisms of these two types of shock. Therefore, the NOx levels reduction by DOX indicates important protection for the injuries associated with HS and are in accordance with previous studies that showed the potential of DOX treatment to decrease NOx levels both by reducing the activity and expression of NOS-2 [12, 13].

Moreover, NOx levels were positively linked with the indicators of renal dysfunction, urea and creatinine, indicating some level of connection. However, no correlation was observed with the indicators of liver injury, AST and ALT, which suggests alternative protective mechanisms involving these markers that are not yet understood.

Some limitations of the study should be mentioned. First, the use of anaesthetic sodium thiopenthone. Anaesthetics used in animal experimentation interferes with several parameters. Barbiturates may have effects on physiological parameters and the recovery is marked by a prolonged period before full consciousness is restored. Therefore, the use of different anaesthetics imposes limitations when comparing studies [29]. Another limitation was the use of crystalloid during resuscitation. Although we used the shed blood for resuscitation, a small amount of crystalloid was also administrated. There is a long discussion about what is the best fluid for resuscitation. Balanced crystalloid offers theoretic benefits over normal saline; however, large studies demonstrate only minimal benefits [30]. A recent meta-analysis pointed that among sepsis and surgical patients, balanced crystalloid and albumin attained lower mortality rates and lower risk of acute kidney injury than saline and low molecular weight hydroxyethyl starch. However, balanced crystalloids required the greatest fluid resuscitation volume than all other fluid types, and there are clear harms associated with the aggressive use of fluid administration [31]. Therefore, the choice of the anaesthetics and the fluids may interfere with the outcome and our results should be interpreted with caution when comparing to other studies.

Conclusion

In summary, our findings show that DOX protected liver and kidney against injury and dysfunction in a relevant HS model and could be a strategy to reduce organ damage associated with ischemia-and-reperfusion injury.

Availability of data and materials

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

References

  1. Haagsma JA, Graetz N, Bolliger I, Naghavi M, Higashi H, Mullany EC et al (2016) The global burden of injury: incidence, mortality, disability-adjusted life years and time trends from the Global Burden of Disease study 2013. Inj Prev 22(1):3–18. https://doi.org/10.1136/injuryprev-2015-041616

    Article  PubMed  Google Scholar 

  2. Cannon JW (2018) Hemorrhagic shock. N Engl J Med 378(4):370–379. https://doi.org/10.1056/NEJMra1705649

    Article  PubMed  Google Scholar 

  3. de Souza P, Schulz R, da Silva-Santos JE (2015) Matrix metalloproteinase inhibitors prevent sepsis-induced refractoriness to vasoconstrictors in the cecal ligation and puncture model in rats. Eur J Pharmacol 765:164–170. https://doi.org/10.1016/j.ejphar.2015.08.030

    Article  CAS  PubMed  Google Scholar 

  4. Wu F, Dorman B, Zeineddin A, Kozar RA (2023) Fibrinogen inhibits metalloproteinase-9 activation and syndecan-1 cleavage to protect lung function in ApoE null mice after hemorrhagic shock. J Surg Res 288:208–214. https://doi.org/10.1016/j.jss.2023.02.043

    Article  CAS  PubMed  Google Scholar 

  5. Pushpakom S, Iorio F, Eyers PA, Escott KJ, Hopper S, Wells A, Doig A et al (2019) Drug repurposing: progress, challenges and recommendations. Nat Rev Drug Discov 18(1):41–58. https://doi.org/10.1038/nrd.2018.168

    Article  CAS  PubMed  Google Scholar 

  6. Xu DQ, Gao C, Niu W, Li Y, Wang YX, Gao CJ et al (2013) Sodium hydrosulfide alleviates lung inflammation and cell apoptosis following resuscitated hemorrhagic shock in rats. Acta Pharmacol Sin 34(12):1515–1525. https://doi.org/10.1038/aps.2013.96

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Sordi R, Nandra KK, Chiazza F, Johnson FL, Cabrera CP, Torrance HD et al (2017) Artesunate protects against the organ injury and dysfunction induced by severe hemorrhage and resuscitation. Ann Surg 265(2):408–417. https://doi.org/10.1097/SLA.0000000000001664

    Article  PubMed  Google Scholar 

  8. Sordi R, Chiazza F, Collotta D, Migliaretti G, Colas RA, Vulliamy P et al (2021) Resolvin D1 attenuates the organ injury associated with experimental hemorrhagic shock. Ann Surg 273(5):1012–1021. https://doi.org/10.1097/SLA.0000000000003407

    Article  PubMed  Google Scholar 

  9. Lu Y, Shimizu H, Nakamura R, Li Y, Sakamoto R, Omori E et al (2023) Dexmedetomidine improves acute lung injury by activating autophagy in a rat hemorrhagic shock and resuscitation model. Sci Rep 13(1):4374. https://doi.org/10.1038/s41598-023-31483-1

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Cerisano G, Buonamici P, Valenti R, Sciagrà R, Raspanti S, Santini A et al (2014) Early short-term doxycycline therapy in patients with acute myocardial infarction and left ventricular dysfunction to prevent the ominous progression to adverse remodelling: the TIPTOP trial. Eur Heart J 35(3):184–191. https://doi.org/10.1093/eurheartj/eht420

    Article  CAS  PubMed  Google Scholar 

  11. Clemens DL, Duryee MJ, Hall JH, Thiele GM, Mikuls TR, Klassen LW et al (2020) Relevance of the antioxidant properties of methotrexate and doxycycline to their treatment of cardiovascular disease. Pharmacol Ther 205:107413. https://doi.org/10.1016/j.pharmthera.2019.107413

    Article  CAS  PubMed  Google Scholar 

  12. D’Agostino P, La Rosa M, Barbera C, Arcoleo F, Di Bella G, Milano S et al (1998) Doxycycline reduces mortality to lethal endotoxemia by reducing nitric oxide synthesis via an interleukin-10-independent mechanism [published correction appears in J Infect Dis 1998 Jun; 177(6):1778]. J Infect Dis 177(2):489–492. https://doi.org/10.1086/517383

    Article  CAS  PubMed  Google Scholar 

  13. Amin AR, Attur MG, Thakker GD, Patel PD, Vyas PR, Patel RN et al (1996) A novel mechanism of action of tetracyclines: effects on nitric oxide synthases. Proc Natl Acad Sci U S A 93(24):14014–14019. https://doi.org/10.1073/pnas.93.24.14014

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Stechmiller J, Cowan L, Schultz G (2010) The role of doxycycline as a matrix metalloproteinase inhibitor for the treatment of chronic wounds. Biol Res Nurs 11(4):336–344. https://doi.org/10.1177/1099800409346333

    Article  CAS  PubMed  Google Scholar 

  15. Gomes JR, Omar NF, Neves JDS, Novaes PD (2017) Doxycycline reduces the expression and activity of matrix metalloproteinase-2 in the periodontal ligament of the rat incisor without altering the eruption process. J Periodontal Res 52(3):353–359. https://doi.org/10.1111/jre.12398

    Article  CAS  PubMed  Google Scholar 

  16. O’Dell JR, Elliott JR, Mallek JA, Mikuls TR, Weaver CA, Glickstein S et al (2006) Treatment of early seropositive rheumatoid arthritis: doxycycline plus methotrexate versus methotrexate alone. Arthritis Rheum 54(2):621–627. https://doi.org/10.1002/art.21620

    Article  CAS  PubMed  Google Scholar 

  17. Yağan A, Kesim S, Liman N (2014) Effect of low-dose doxycycline on serum oxidative status, gingival antioxidant levels, and alveolar bone loss in experimental periodontitis in rats. J Periodontol 85(3):478–489. https://doi.org/10.1902/jop.2013.130138

    Article  CAS  PubMed  Google Scholar 

  18. Patel A, Khande H, Periasamy H, Mokale S (2020) Immunomodulatory effect of doxycycline ameliorates systemic and pulmonary inflammation in a murine polymicrobial sepsis model. Inflammation 43(3):1035–1043. https://doi.org/10.1007/s10753-020-01188-y

    Article  CAS  PubMed  Google Scholar 

  19. Sordi R, Chiazza F, Collino M, Assreuy J, Thiemermann C (2016) Neuronal nitric oxide synthase is involved in vascular hyporeactivity and multiple organ dysfunction associated with hemorrhagic shock. Shock 45(5):525–533. https://doi.org/10.1097/SHK.0000000000000533

    Article  CAS  PubMed  Google Scholar 

  20. Sordi R, Chiazza F, Patel NS, Doyle RA, Collino M, Thiemermann C (2015) ‘Preconditioning’ with low dose lipopolysaccharide aggravates the organ injury / dysfunction caused by hemorrhagic shock in rats. PLoS ONE 10(4):e0122096. https://doi.org/10.1371/journal.pone.0122096

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Eltzschig HK, Eckle T (2011) Ischemia and reperfusion–from mechanism to translation. Nat Med 17(11):1391–1401. https://doi.org/10.1038/nm.2507

    Article  CAS  PubMed  Google Scholar 

  22. Kholmukhamedov A, Czerny C, Hu J, Schwartz J, Zhong Z, Lemasters JJ (2014) Minocycline and doxycycline, but not tetracycline, mitigate liver and kidney injury after hemorrhagic shock/resuscitation. Shock 42(3):256–263. https://doi.org/10.1097/SHK.0000000000000213

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Schwartz J, Holmuhamedov E, Zhang X, Lovelace GL, Smith CD, Lemasters JJ (2013) Minocycline and doxycycline, but not other tetracycline-derived compounds, protect liver cells from chemical hypoxia and ischemia/reperfusion injury by inhibition of the mitochondrial calcium uniporter. Toxicol Appl Pharmacol 273(1):172–179. https://doi.org/10.1016/j.taap.2013.08.027

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Fernandes D, Assreuy J (2008) Nitric oxide and vascular reactivity in sepsis. Shock 30(Suppl 1):10–13. https://doi.org/10.1097/SHK.0b013e3181818518

    Article  CAS  PubMed  Google Scholar 

  25. Sordi R, Fernandes D, Assreuy J (2010) Differential involvement of potassium channel subtypes in early and late sepsis-induced hyporesponsiveness to vasoconstrictors. J Cardiovasc Pharmacol 56(2):184–189. https://doi.org/10.1097/FJC.0b013e3181e74d6a

    Article  CAS  PubMed  Google Scholar 

  26. Sordi R, Fernandes D, Heckert BT, Assreuy J (2011) Early potassium channel blockade improves sepsis-induced organ damage and cardiovascular dysfunction. Br J Pharmacol 163(6):1289–1301. https://doi.org/10.1111/j.1476-5381.2011.01324.x

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Okamoto I, Abe M, Shibata K, Shimizu N, Sakata N, Katsuragi T, Tanaka K (2000) Evaluating the role of inducible nitric oxide synthase using a novel and selective inducible nitric oxide synthase inhibitor in septic lung injury produced by cecal ligation and puncture. Am J Respir Crit Care Med 162(2 Pt 1):716–722. https://doi.org/10.1164/ajrccm.162.2.9907039

    Article  CAS  PubMed  Google Scholar 

  28. Thiemermann C, Szabó C, Mitchell JA, Vane JR (1993) Vascular hyporeactivity to vasoconstrictor agents and hemodynamic decompensation in hemorrhagic shock is mediated by nitric oxide. Proc Natl Acad Sci U S A 90(1):267–271. https://doi.org/10.1073/pnas.90.1.267

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Tobar Leitão SA, Soares DDS, Carvas Junior N, Zimmer R, Ludwig NF, Andrades M (2021) Study of anesthetics for euthanasia in rats and mice: a systematic review and meta-analysis on the impact upon biological outcomes (SAFE-RM). Life Sci 1(284):119916. https://doi.org/10.1016/j.lfs.2021.119916

    Article  CAS  Google Scholar 

  30. Gordon D, Spiegel R (2020) Fluid resuscitation: history, physiology, and modern fluid resuscitation strategies. Emerg Med Clin North Am 38(4):783–793. https://doi.org/10.1016/j.emc.2020.06.004

    Article  PubMed  Google Scholar 

  31. Tseng CH, Chen TT, Wu MY, Chan MC, Shih MC, Tu YK (2020) Resuscitation fluid types in sepsis, surgical, and trauma patients: a systematic review and sequential network meta-analyses. Crit Care 24(1):693. https://doi.org/10.1186/s13054-020-03419-y

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

The authors are grateful to the Multiuser Laboratory of Biology Studies at the Universidade Federal de Santa Catarina (LAMEB/UFSC) for providing infrastructure for carrying out the experimental tests and to Adriane Madeira for the technical assistance.

Funding

This study was supported by National Council for Scientific and Technological Development (CNPq; grant number 409018/2018-0; 405538/2021-9 and 403615/2023-2) and Coordination for the Improvement of Higher Education Personnel (CAPES Foundation), Brazil.

Author information

Authors and Affiliations

Authors

Contributions

RS and JRG were involved in the conception, hypotheses delineation and design of the study; LB, RS, FRMBO, TOR, CFS, LWM, GFA were involved in the acquisition of the data and interpretation of such information. RS, DF, JRG and JCRV were involved in data analysis, writing the article or substantial involvement in its revision prior to submission. All the authors revised and approved the final version of the article.

Corresponding author

Correspondence to Jose Rosa Gomes.

Ethics declarations

Ethics approval and consent to participate

The procedures were approved by the Universidade Estadual de Ponta Grossa Ethics Committee (protocol number 048/2015).

Consent for publication

The authors affirm that they have no conflicting interests in publishing this manuscript.

Competing interests

The authors declare no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sordi, R., Bojko, L., Oliveira, F.R.M.B. et al. Doxycycline reduces liver and kidney injuries in a rat hemorrhagic shock model. ICMx 12, 2 (2024). https://doi.org/10.1186/s40635-023-00586-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1186/s40635-023-00586-4

Keywords