Log in | Register

Endotoxin-induced gene expression differences in the brain and effects of iNOS inhibition and norepinephrineOpen access

Stephanie Wolff| Sabine Klatt| Jens C. Wolff| Jochen Wilhelm| Ludger Fink| Manfred Kaps| Bernhard Rosengarten
Experimental
Volume 35, Issue 4 / April , 2009

Pages 730 - 739

Abstract

Purpose

We studied gene expression differences in brain homogenate, hippocampus, somatosensory cortex and cerebellum of rats suffering from sepsis-associated delirium and analyzed the effects of norepinephrine and 1,400 W (specific inhibitor of the inducible nitric-oxide synthase).

Methods

We applied microarray screenings to rat brain homogenate 1, 3 and 4.5 h after lipopolysaccharide (LPS, 5 mg/kg) or 0.9% NaCl treatment. Therapy groups were analyzed after 4.5 h. Validations and compartment specific investigations were carried out by real-time PCR.

Results

Most striking gene expression differences were seen 4.5 h after LPS administration, especially within the hippocampus (chemokines and endothelial cell-specific molecule 1). Norepinephrine resulted in a discrete chemokine up-regulation, while 1,400 W had hardly any effect.

Conclusion

Strongest gene regulations were found within the hippocampus. Norepinephrine showed a tendency of having a proinflammatory influence, while 1,400 W had no clear-cut effect onto the gene expression level.

Keywords

References

  1. Ebersoldt M, Sharshar T, Annane D (2007) Sepsis-associated delirium. Intensive Care Med 33:941–950
  2. Green R, Scott LK, Minagar A, Conrad S (2004) Sepsis associated encephalopathy (SAE): a review. Front Biosci 9:1637–1641
    • View reference on PubMed
    • View reference on publisher's website
  3. Papadopoulos MC, Davies DC, Moss RF, Tighe D, Bennett ED (2000) Pathophysiology of septic encephalopathy: a review. Crit Care Med 28:3019–3024
    • View reference on PubMed
    • View reference on publisher's website
  4. Bello JH, Park M (2007) Sepsis-associated encephalopathy as a differential diagnosis with motor deficit plus altered mental status. Clinics 62:199–202
    • View reference on PubMed
    • View reference on publisher's website
  5. Eggers V, Schilling A, Kox WJ, Spies C (2003) Septic encephalopathy. Diagnosis und therapy. Anaesthesist 52:294–303
    • View reference on PubMed
    • View reference on publisher's website
  6. Sharshar T, Annane D, de la Grandmaison GL, Brouland JP, Hopkinson NS, Françoise G (2004) The neuropathology of septic shock. Brain Pathol 14:21–33
    • View reference on PubMed
  7. Hinkelbein J, Schroeck H, Peterka A, Schubert C, Kuschinsky W, Kalenka A (2007) Local cerebral blood flow is preserved in sepsis. Curr Neurovasc Res 4:39–47
    • View reference on PubMed
    • View reference on publisher's website
  8. Pfister D, Siegemund M, Dell-Kuster S, Smielewski P, Rüegg S, Strebel SP, Marsch SC, Pargger H, Steiner LA (2008) Cerebral perfusion in sepsis-associated delirium. Crit Care 12:R63
    • View reference on PubMed
    • View reference on publisher's website
  9. Lee CC, Lin NT, Hsu YH, Chen HI (2005) Inducible nitric oxide synthase inhibition potentiates multiple organ dysfunction induced by endotoxin in conscious rats. J Cardiovasc Pharmacol 9:139–143
  10. Matejovic M, Krouzecky A, Radej J, Rokyta R Jr, Kralova H, Radermacher P, Novak I (2007) Coagulation and endothelial dysfunction during longterm hyperdynamic porcine bacteremia-effects of selective inducible nitric oxide synthase inhibition. Thromb Haemost 97:304–309
    • View reference on PubMed
  11. Ahn KS, Noh EJ, Zhao HL, Jung SH, Kang SS, Kim YS (2005) Inhibition of inducible nitric oxide synthase and cyclooxygenase II by Platycodon grandiflorum saponins via suppression of nuclear factor-kappaB activation in RAW 264.7 cells. Life Sci 76:2315–2328
    • View reference on PubMed
    • View reference on publisher's website
  12. Rosengarten B, Hecht M, Auch D, Ghofrani HA, Schermuly RT, Grimminger F, Kaps M (2007) Microcirculatory dysfunction in the brain precedes changes in evoked potentials in endotoxin-induced sepsis syndrome in rats. Cerebrovasc Dis 23:140–147
    • View reference on PubMed
    • View reference on publisher's website
  13. R Development Core Team (2007) R: a language and environment for statistical computing. R Foundation for statistical computing, Vienna, Austria. ISBN 3-900051-07-0. Available via DIALOG http://www.R-project.org. Accessed 2007
  14. Smyth GK (2005) Limma: linear models for microarray data. In: Gentleman R, Carey V, Dudoit S, Irizarry R, Huber W (eds) Bioinformatics and computational biology solutions using R and bioconductor. Springer, New York, pp 397–420
  15. Gentleman RC, Carey VJ, Bates DM, Bolstad B, Dettling M, Dudoit S, Ellis B, Gautier L, Ge Y, Gentry J, Hornik K, Hothorn T, Huber W, Iacus S, Irizarry R, Leisch F, Li C, Maechler M, Rossini AJ, Sawitzki G, Smith C, Smyth G, Tierney L, Yang JY, Zhang J (2004) Bioconductor: open software development for computational biology and bioinformatics. In: Genome Biol 5:R80. Available via DIALOG http://genomebiology.com/2004/5/10/R80. Accessed 2004
  16. Edwards D (2003) Non-linear normalization and background correction in one-channel cDNA microarray studies. Bioinformatics 19:825–833
    • View reference on PubMed
    • View reference on publisher's website
  17. Smyth GK, Speed TP (2003) Normalization of cDNA microarray data. Methods 31:265–273
    • View reference on PubMed
    • View reference on publisher's website
  18. Smyth GK (2004) Linear models and empirical Bayes methods for assessing differential expression in microarray experiments. In: Statistical applications in genetics and molecular biology 3, article 3. Available via DIALOG http://www.bepress.com/sagmb/vol3/iss1/art3. Accessed 2004
  19. Benjamini Y, Hochberg Y (1995) Controlling the false discovery rate: a practical and powerful approach to multiple testing. J R Stat Soc Ser B 57:289–300
  20. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408
    • View reference on PubMed
    • View reference on publisher's website
  21. Fink L, Kohlhoff S, Stein MM, Hänze J, Weissmann N, Rose F, Akkayagil E, Manz D, Grimminger F, Seeger W, Bohle RM (2002) cDNA array hybridization after laser-assisted microdissection from nonneoplastic tissue. Am J Pathol 160:81–90
    • View reference on PubMed
  22. Silverman J, Muir WW 3rd (1993) A review of laboratory animal anesthesia with chloral hydrate and chloralose. Lab Anim Sci 43:210–216
    • View reference on PubMed
  23. Wilhelm J, Muyal JP, Best J, Kwapiszewska G, Stein MM, Seeger W, Bohle RM, Fink L (2006) Systematic comparison of the T7-IVT and SMART-based RNA preamplification techniques for DNA microarray experiments. Clin Chem 52:1161–1167
    • View reference on PubMed
    • View reference on publisher's website
  24. Schneider J, Buness A, Huber W, Volz J, Kioschis P, Hafner M, Poustka A, Sültmann H (2004) Systematic analysis of T7 RNA polymerase based in vitro linear RNA amplification for use in microarray experiments. BMC Genomics 5:29
    • View reference on PubMed
    • View reference on publisher's website
  25. Puskás LG, Zvara A, Hackler L Jr, Van Hummelen P (2002) RNA amplification results in reproducible microarray data with slight ratio bias. Biotechniques 32:1330–1334, 1336, 1338, 1340
  26. Carrol ED, Mankhambo LA, Balmer P, Nkhoma S, Banda DL, Guiver M, Jeffers G, Makwana N, Molyneux EM, Molyneux ME, Smyth RL, Hart CA, Invasive Pneumococcal Disease Study Group (2007) Chemokine responses are increased in HIV-infected Malawian children with invasive pneumococcal disease. J Acquir Immune Defic Syndr 44:443–450
    • View reference on PubMed
    • View reference on publisher's website
  27. Riese J, Schoolmann S, Denzel C, Herrmann O, Hohenberger W, Haupt W (2002) Effect of abdominal infections on peritoneal and systemic production of interleukin 6 and monocyte chemoattractant protein-1. Shock 17:361–364
    • View reference on PubMed
    • View reference on publisher's website
  28. Bossink AW, Paemen L, Jansen PM, Hack CE, Thijs LG, Van Damme J (1995) Plasma levels of the chemokines monocyte chemotactic proteins-1 and -2 are elevated in human sepsis. Blood 86:3841–3847
    • View reference on PubMed
  29. Kalehua AN, Nagel JE, Whelchel LM, Gides JJ, Pyle RS, Smith RJ, Kusiak JW, Taub DD (2004) Monocyte chemoattractant protein-1 and macrophage inflammatory protein-2 are involved in both excitotoxin-induced neurodegeneration and regeneration. Exp Cell Res 297:197–211
    • View reference on PubMed
    • View reference on publisher's website
  30. Eugenin EA, D’Aversa TG, Lopez L, Calderon TM, Berman JW (2003) MCP-1 (CCL2) protects human neurons and astrocytes from NMDA or HIV-tat-induced apoptosis. J Neurochem 85:1299–1311
    • View reference on PubMed
    • View reference on publisher's website
  31. Dimitrijevic OB, Stamatovic SM, Keep RF, Andjelkovic AV (2007) Absence of the chemokine receptor CCR2 protects against cerebral ischemia/reperfusion injury in mice. Stroke 38:1345–1353
    • View reference on PubMed
    • View reference on publisher's website
  32. Abraham E, Kaneko DJ, Shenkar R (1999) Effects of endogenous and exogenous catecholamines on LPS-induced neutrophil trafficking and activation. Am J Physiol 276:L1–L8
    • View reference on PubMed
  33. Hofstetter C, Boost KA, Hoegl S, Flondor M, Scheller B, Muhl H, Pfeilschifter J, Zwissler B (2007) Norepinephrine and vasopressin counteract anti-inflammatory effects of isoflurane in endotoxemic rats. Int J Mol Med 20:597–604
    • View reference on PubMed
  34. Tsuda Y, Kobayashi M, Herndon DN, Suzuki F (2008) Impairment of the host’s antibacterial resistance by norepinephrine activated neutrophils. Burns 34:460–466
    • View reference on PubMed
    • View reference on publisher's website
  35. Blandino P Jr, Barnum CJ, Deak T (2006) The involvement of norepinephrine and microglia in hypothalamic and splenic IL-1beta responses to stress. J Neuroimmunol 173:87–95
    • View reference on PubMed
    • View reference on publisher's website
  36. Conte D, Holcik M, Lefebvre CA, Lacasse E, Picketts DJ, Wright KE, Korneluk RG (2006) Inhibitor of apoptosis protein cIAP2 is essential for lipopolysaccharide-induced macrophage survival. Mol Cell Biol 26:699–708
    • View reference on PubMed
    • View reference on publisher's website
  37. Tissier S, Lancel S, Marechal X, Mordon S, Depontieu F, Scherpereel A, Chopin C, Neviere R (2004) Calpain inhibitors improve myocardial dysfunction and inflammation induced by endotoxin in rats. Shock 21:352–357
    • View reference on PubMed
    • View reference on publisher's website
  38. Scherpereel A, Depontieu F, Grigoriu B, Cavestri B, Tsicopoulos A, Gentina T, Jourdain M, Pugin J, Tonnel AB, Lassalle P (2006) Endocan, a new endothelial marker in human sepsis. Crit Care Med 34:532–537
    • View reference on PubMed
    • View reference on publisher's website
  39. Semmler A, Okulla T, Sastre M, Dumitrescu-Ozimek L, Heneka MT (2005) Systemic inflammation induces apoptosis with variable vulnerability of different brain regions. J Chem Neuroanat 30:144–157
    • View reference on PubMed
    • View reference on publisher's website

Sign In

Connect with ICM

Top 5 Articles Editors Picks Supplement