Journal name: Journal of Inflammation Research Article Designation: Original Research Year: 2018 Volume: 11 Journal of Inflammation Research Dovepress Running head verso: Jägers et al Running head recto: Moderate glucose supply reduces hemolysis open access to scientific and medical research DOI: http://dx.doi.org/10.2147/JIR.S155614 Open Access Full Text Article ORIgInal ReseaRch Moderate glucose supply reduces hemolysis during systemic inflammation Johannes Jägers1 Background: Systemic inflammation alters energy metabolism. A sufficient glucose level, stephan Brauckmann2 however, is most important for erythrocytes, since erythrocytes rely on glucose as sole source Michael Kirsch1 of energy. Damage to erythrocytes leads to hemolysis. Both disorders of glucose metabolism Katharina and hemolysis are associated with an increased risk of death. The objective of the study was effenberger-neidnicht1,3 to investigate the impact of intravenous glucose on hemolysis during systemic inflammation. Materials and methods: Systemic inflammation was accomplished in male Wistar rats by 1 Institute of Physiological chemistry, continuous lipopolysaccharide (LPS) infusion (1 mg LPS/kg and h, 300 min). Sham control University hospital essen, essen, germany; 2clinic for anesthesiology group rats received Ringer’s solution. Glucose was supplied moderately (70 mg glucose/kg and and Intensive care, University h) or excessively (210 mg glucose/kg and h) during systemic inflammation. Vital parameters hospital essen, essen, germany; For personal use only. 3Institute of Physiological chemistry, (eg, systemic blood pressure) as well as blood and plasma parameters (eg, concentrations of University hospital essen, essen, glucose, lactate and cell-free hemoglobin, and activity of lactate dehydrogenase) were measured germany hourly. Clot formation was analyzed by thromboelastometry. Results: Continuous infusion of LPS led to a so-called post-aggression syndrome with disturbed electrolyte homeostasis (hypocalcemia, hyperkalemia, and hypernatremia), changes in hemody- namics (tachycardia and hypertension), and a catabolic metabolism (early hyperglycemia, late hypoglycemia, and lactate formation). It induced severe tissue injury (significant increases in plasma concentrations of transaminases and lactate dehydrogenase), alterations in blood coagula- tion (disturbed clot formation), and massive hemolysis. Both moderate and excessive glucose sup- ply reduced LPS-induced increase in systemic blood pressure. Excessive but not moderate glucose supply increased blood glucose level and enhanced tissue injury. Glucose supply did not reduce LPS-induced alterations in coagulation, but significantly reduced hemolysis induced by LPS. Journal of Inflammation Research downloaded from https://www.dovepress.com/ by 54.70.40.11 on 20-Dec-2018 Conclusion: Intravenous glucose infusion can diminish LPS-related changes in hemody- namics, glucose metabolism, and, more interestingly, LPS-induced hemolysis. Since cell-free hemoglobin is known to be a predictor for patient’s survival, a reduction of hemolysis by 35% only by the addition of a small amount of glucose is another step to minimize mortality during systemic inflammation. Keywords: lipopolysaccharide, sepsis, erythrocytes, red blood cells, cell-free hemoglobin, glucose metabolism correspondence: Katharina Introduction effenberger-neidnicht Blood glucose level is tightly regulated in healthy individuals.1,2 In situations of critical Institute of Physiological chemistry, illness (eg, trauma, burn, or systemic inflammation), however, energy metabolism and University hospital essen, hufelandstraße 55, essen D-45122, germany glucose homeostasis are dysregulated.1–6 The metabolic response to injury and sepsis is Tel +49 201 723 4103 often called auto-cannibalism and includes a rapid mobilization of all substrate depots email katharina.effenberger-neidnicht@ uni-due.de (eg, glycogen) and, accordingly, the loss of muscle mass and functional proteins, body submit your manuscript | www.dovepress.com Journal of Inflammation Research 2018:11 87–94 87 Dovepress © 2018 Jägers et al. This work is published and licensed by Dove Medical Press Limited. 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Powered by TCPDF (www.tcpdf.org) 1 / 1 Jägers et al Dovepress fat, and the glucose storage.3–7 During systemic inflamma- from AbbVie Deutschland (Ludwigshafen, Germany), ket- tion, higher glucose requirement is covered by both increased amine 10% from Ceva (Düsseldorf, Germany), lidocaine hepatic glycogenolysis and gluconeogenesis leading to an (Xylocaine 1%) from AstraZeneca (Wedel, Germany), medi- initial hyperglycemic state.5,6,8 However, the glycogen depots cal oxygen from Air Liquide (Düsseldorf, Germany), Ringer’s are quickly exhausted after some time in case of higher solution from Fresenius Kabi (Bad Homburg, Germany), glucose consumption. From this moment onward, glucose and 2.0 mL self-filling arterial samplers containing 80 IU supply can only be achieved by an increased gluconeogenesis electrolyte-balanced heparin (PICO50) from Radiometer in the liver (temporary euglycemic phase in metabolic stress Medical (Brønshøj, Denmark). Portex catheters (0.58 mm response).5,6,8 If this possibility of endogenous glucose pro- i.d./0.96 mm o.d.) were obtained from Smith Medical Inter- duction is also restricted or if the glucose uptake rate exceeds national (Grasbrunn, Germany). the rate of glucose production – as it is the case for severe sepsis and inflammation – finally hypoglycemia occurs.5,6,8 animals Clinical studies showed that hypoglycemia or hyperglycemia Male Wistar rats (390–520 g, from Envigo, formerly (defined as malglycemia) is associated with an increased risk Harlan Laboratories, Horst, Netherlands) received human of death.9–12 In situation of critical illness, glucose as energy care according to standards of the Federation of European source for the brain is replaced almost completely by ketone Laboratory Animal Science Association. The experimental bodies.6 Erythrocytes, however, are dependent on glucose procedure was reviewed and approved by the local Animal as the sole source of energy (anaerobic glycolysis),13,14 Care and Use Committee with the permit number Az. even though under catabolic conditions, for a short time, 84-02.04.2013.A015 (Landesamt für Natur, Umwelt und the metabolism of 2,3-diphosphoglycerate to pyruvate and Verbraucherschutz Nordrhein-Westfalen, Recklinghausen, lactate is used for adenosine triphosphate (ATP) synthesis Germany). Rats were kept under standardized conditions of as well.15 “Starved” erythrocytes perish and hemoglobin and temperature (22 ± 1°C), humidity (55 ± 5%), and 12-h/12-h other cytoplasmic components, eg, lactate dehydrogenase light/dark cycles (06:00 a.m. light on/06:00 p.m. light off, For personal use only. and potassium, are released (so-called hemolysis).14 Cell-free standard time) with free access to food (ssniff-Spezialdiäten, hemoglobin and heme can contribute to organ dysfunction Soest, Germany) and water. and death;16–19 the pathologic mechanisms include nitric oxide (NO) consumption and local vasoconstriction, oxida- anesthesia, analgesia and surgical tive injury to lipid membranes, activation of the transcription procedure factor NF-κB, endothelial injury, as well as iron-driven oxida- Rats were anesthetized with isoflurane and received ket- tive inhibition of glucose metabolism.19–22 Thus, hemolysis amine and lidocaine for analgesia as already described.30 can act as a kind of amplifier of the complex response to an Afterwards, catheters were inserted in femoral artery and infection or injury23,24 and worsen the outcome in patients vein and perfused with Ringer’s solution to maintain the with sepsis or trauma.16–19 In contrast to other studies, glucose functionality of the catheters. At the end of the experiment, 9,25,26 Journal of Inflammation Research downloaded from https://www.dovepress.com/ by 54.70.40.11 on 20-Dec-2018 balance was not performed via insulin treatment, but via animals were sacrificed by cardiac incision under deep iso- exogenous glucose supply.27–29 Blood glucose level should be flurane anesthesia. kept high, especially during hypoglycemia, and consequences such as hemolysis through glucose depletion in erythrocytes experimental groups should be avoided. Experiments were performed with overall 40 rats. Sham control In the present manuscript we wanted to investigate group rats received Ringer’s solution at a rate of 7 mL/kg and whether lipopolysaccharide (LPS)-induced hemolysis can h over a total period of 300 min. Systemic inflammation was be prevented or reduced if more glucose was present as a accomplished by continuous LPS infusion over a total period substrate for erythrocytes. of 300 min. Glucose was supplied either moderately or exces- sively during systemic inflammation simultaneously with the Materials and methods LPS infusion. During moderate glucose supply, rats received chemicals and materials LPS in