Abstract
Clinical and experimental evidence indicate that infection in pregnancy is associated with fetal brain damage. However, the inflammatory processes that compromise the fetal brain are not fully understood. In this study, we used a single, low dose of lipopolysaccharide (LPS, 0.1 μg/kg i.v.) to provoke an acute-phase response in unanesthetized fetal sheep in utero. COX-2 mRNA was increased in the cortex and cerebellum at 24 and 48 h after LPS, and immunoreactive COX-2 protein was increased in perivascular cells throughout gray and white matter at 24 h after LPS administration. Plasma albumin was observed in the parenchyma of the brain in cortex, thalamus, hypothalamus, corpus callosum, fornix, hippocampus, midbrain, subcallosal bundle, and cerebellar Purkinje cells. Large, rounded, lectin-positive cells with the appearance of macrophages were observed around blood vessels in subventricular white matter. These results indicate that blood-brain barrier permeability is increased in the fetal brain after exposure to endotoxin and suggests that cytotoxic and pro-inflammatory substances could pass from the circulation into the brain after peripheral inflammatory stimulation.
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Abbreviations
- BBB:
-
blood-brain barrier
- COX-1:
-
cyclooxygenase type-1
- COX-2:
-
cyclooxygenase type-2
- GAPDH:
-
glyceraldehyde 3-phophate dehydrogenase
- iNOS:
-
inducible nitric oxide synthase
- LPS:
-
lipopolysaccharide
- NO:
-
nitric oxide
- PB:
-
phosphate buffer
- PG:
-
prostaglandin
- RNase:
-
ribonuclease
- TNF-α:
-
tumor necrosis factor-α
References
Grether JK, Nelson KB 1997 Maternal infection and cerebral palsy in infants of normal birth weight. JAMA 278: 207–211.
Van Hoeven KH, Anyaegbunam A, Hochster H, Whitty JE, Distant J, Crawford C, Factor SM 1996 Clinical significance of increasing histologic severity of acute inflammation in the fetal membranes and umbilical cord. Pediatr Pathol Lab Med 16: 731–744.
Baud O, Emilie D, Pelletier E, Lacaze-Masmonteil T, Zupan V, Fernandez H, Dehan M, Frydman R, Ville Y 1999 Amniotic fluid concentrations of interleukin-1beta, interleukin-6 and TNF-α in chorioamnionitis before 32 weeks of gestation: histological associations and neonatal outcome. Br J Obstet Gynaecol 106: 72–77.
Kim CJ, Yoon BH, Romero R, Moon JB, Kim M, Park S-S, Chi JG 2001 Umbilical arteritis and phlebitis mark different stages of the inflammatory response. Am J Obstet Gynecol 185: 496–500.
Dammann O, Leviton A 1998 Infection remote from the brain, neonatal white matter damage, and cerebral palsy in the preterm infant. Semin Pediatr Neurol 5: 190–201.
Elmquist JK, Breder CD, Sherin JE, Scammell TE, Hickey WF, Dewitt D, Saper CB 1997 Intravenous lipopolysaccharide induces cyclooxygenase 2-like immunoreactivity in rat brain perivascular microglia and meningeal macrophages. J Comp Neurol 381: 119–129.
Banks WA, Kastin AJ, Durham DA 1989 Bidirectional transport of interleukin-1 alpha across the blood-brain barrier. Brain Res Bull 23: 433–437.
Engblom D-D 2002 Prostaglandins as inflammatory messengers across the blood-brain barrier. J Mol Med 80: 5–15.
Moore SA, Spector AA, Hart MN 1988 Eicosanoid metabolism in cerebromicrovascular endothelium. Am J Physiol 254:C37–C44.
Cao C, Matsumura K, Yamagata K, Watanabe Y 1995 Induction by lipopolysaccharide of cyclooxygenase-2 mRNA in rat brain; its possible role in the febrile response. Brain Res 697: 187–196.
Elmquist JK, Scammell TE, Saper CB 1997 Mechanisms of CNS response to systemic immune challenge: the febrile response. Trends Neurosci 20: 565–570.
Kennedy BP, Chan CC, Culp SA, Cromlish WA 1993 Cloning and expression of prostaglandin synthase (cyclooxygenase)-2 cDNA. Biochem Biophys Res Commun 197: 494–500.
Norton JL, Adamson SL, Bocking AD, Han VK 1996 Prostaglandin-H synthase-1 (PGHS-1) gene is expressed in specific neurons of the brain of the late gestation ovine fetus. Brain Res Dev Brain Res 95: 79–96.
Van Dam AM, Brouns M, Man AHW, Berkenbosch F 1993 Immunocytochemical detection of prostaglandin E2 in microvasculature and in neurons of rat brain after administration of bacterial endotoxin. Brain Res 613: 331–336.
Breder CD, Saper CB 1996 Expression of inducible cyclooxygenase mRNA in the mouse brain after systemic administration of bacterial lipopolysaccharide. Brain Res 713: 64–69.
Cao C, Matsumura K, Yamagata K, Watanabe Y 1997 Involvement of cyclooxygenase-2 in LPS-induced fever and regulation of its mRNA by LPS in the rat brain. Am J Physiol 272:R1712–R1725.
Maloney CG, Kutchear WA, Albertine KH, McIntyre TM, Prescott SM, Zimmerman GA 1998 Inflammatory agonist cyclooxygenase type 2 expression by human neutrophils. J Immunol 160: 1402–1410.
Quan N, Whiteside M, Herkenham M 1998 Cyclooxygenase 2 mRNA expression in rat brain after peripheral injection of lipopolysaccharide. Brain Res 802: 189–197.
Yamagata K, Andreasson KI, Kaufmann WE, Barnes CA, Worley PF 1993 Expression of a mitogen-inducible cyclooxygenase in brain neurons: regulation by synaptic activity and glucocorticoids. Neuron 11: 371–386.
Breder CD, Dewitt D, Kraig RP 1995 Characterization of inducible cyclooxygenase in rat brain. J Comp Neurol 355: 296–315.
Tsao N, Hsu H-P, Lei H-Y 1999 TNF-αinduced cyclooxygenase 2 not only increases the vasopermeability of blood-brain barrier but also enhances the neutrophil survival in Escherichia coli-induced brain inflammation. Prostaglandins Other Lipid Mediat 57: 371–382.
Minami T, Okazaki J, Kawabata A, Kawaki H, Okazaki Y, Tohno Y 1998 Roles of nitric oxide and prostaglandins in the increased permeability of the blood-brain barrier caused by lipopolysaccharide. Environ Toxicol Pharmacol 5: 35–41.
Saunders NR 1992 Ontogenetic development of brain barrier mechanisms. In: Bradbury MWB (ed) Handbook of Experimental Pharmacology. Springer-Verlag, Berlin, 327–369.
Saunders NR, Habgood MD, Dziegielewska K 1999 Barrier mechanisms in the brain. II. Immature brain. Clin Exp Pharmacol Physiol 26: 85–91.
Stonestreet BS, Sadowska GB, McKnight AJ, Patlak C, Petersson KH 2000 Exogenous and endogenous corticosteroids modulate blood-brain barrier development in the ovine fetus. Am J Physiol Regul Integr Comp Physiol 279:R468–R477.
Perry VH, Anthony DC, Bolton SJ, Brown HC 1997 The blood-brain barrier and the inflammatory response. Mol Med Today 3: 335–341.
Schlafer DH, Yuh B, Foley TH, Elssaser TH, Sadowsky D, Nathanielsz PW 1994 Effect of salmonella endotoxin administered to the pregnant sheep at 133-142 days gestation on fetal oxygenation, maternal and fetal ACTH and cortisol, and maternal plasma TNF-α concentration. Biol Reprod 50: 1297–1302.
Sehic E, Szekely M, Ungar AL, Oladehin A, Blatteis CM 1996 Hypothalamic prostaglandin E2 during lipopolysaccharide-induced fever in guinea pigs. Brain Res Bull 39: 391–399.
Nicholls T, Nitsos I, Walker DW 1999 Tryptophan metabolism in pregnant sheep: increase fetal kynurenine production in response to maternal tryptophan loading. Am J Obstet Gynecol 181: 1452–1460.
Zhang V, OSullivan M, Hussain H, Roswit WT, Holtzman MJ 1996 Molecular cloning, functional expression and selective regulation of prostaglandin H synthase-2. Biochem Biophys Res Commun 227: 499–506.
Hume DA, Underhill DM, Sweet MJ, Ozinsky AO, Liew FY, Aderem A 2001 Macrophages exposed continuously to lipopolysaccharide and other agonists that act via toll-like receptors exhibit a sustained and additive activation state. BMC Immunol 2: 11
Peri KG, Hardy P, Li DY, Varma DR, Chemtob S 1995 Prostaglandin G/H synthase-2 is a major contributor of brain prostaglandins in the newborn. J Biol Chem 270: 24615–24620.
Parfenova H, Levine V, Gunther WM, Pourcyrous M, Leffler CW 2002 COX-1 and COX-2 contributions to basal and IL-1 beta-stimulated prostanoid synthesis inhuman neonatal cerebral microvascular endothelial cells. Pediatr Res 52: 342–348.
Davidge ST, Baker PN, McLaughlin MK, Roberts JM 1995 Nitric oxide produced by endothelial cells increases production of eicosanoids through activation of prostaglandin H synthase. Circ Res 77: 274–283.
Schlitz JC, Sawchenko PE 2002 Distinct brain vascular cell types manifest inducible cyclooxygenase expression as a function of the strength and nature of immune insults. J Neurosci 22: 5606–5618.
Minghetti L, Walsh DT, Levi G, Perry VH 1999 In vivo expression of cycoloxygenase-2 in rat brain following intraparynchymal injection of bacterial endotoxin and inflammatory cytokines. J Neurpathol Exp Neurol 58: 1184–1191.
Gilles FH, Murphy SF 1969 Perinatal telencephalic leucoencephalopathy. J Neurol Neurosurg Psychiatry 32: 404–413.
Gilles FH, Averill DR, Kerr CS 1977 Neonatal endotoxin encephalopathy. Ann Neurol 2: 49–56.
Skultetyova I, Tokarev D, Jezova D 1998 Stress-induced increase in blood-brain barrier permeability in control and monosodium glutamate-treated rats. Brain Res Bull 45: 175–178.
Dziegielewska KM, Knott GW, Saunders NR 2000 The nature and development of the internal environment of the developing brain. Cell Mol Neurobiol 20: 41–56.
Tabernero A, Granda B, Medina A, Sanchez-Abarca LI, Lavado E, Medina M 2002 Albumin promotes neuronal survival by increasing the synthesis and release of glutamate. J Neurochem 81: 881–891.
Stoll G, Jander S 1999 The role of microglia and macrophages in the pathophysiology of the CNS. Prog Neurobiol 58: 233–247.
Dammann O, Durum S, Leviton A 2001 Do white cells matter in white matter damage? Trends N. eurosci 24: 320–324.
Manuelpillai U, Nicholls T, Wallace EM, Phillips DJ, Guillemin G, Walker D 2003 Increased mRNA expression of kynurenine pathway enzymes in human placentae exposed to bacterial endotoxin. Adv Exp Med Biol 527: 85–89.
Acknowledgements
The authors thank Alex Satragno for his expert help with the surgical procedures. We also thank Dr. K. Dziegielewska for invaluable help and advice with the immunohistochemistry and histology.
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Supported by a Monash University Postgraduate Scholarship (E.Y., T.N.) and a National Health and Medical Research Council of Australia grant (D.W., J.H., M.C.M.).
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Yan, E., Castillo-Meléndez, M., Nicholls, T. et al. Cerebrovascular Responses in the Fetal Sheep Brain to Low-Dose Endotoxin. Pediatr Res 55, 855–863 (2004). https://doi.org/10.1203/01.PDR.0000115681.95957.D4
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DOI: https://doi.org/10.1203/01.PDR.0000115681.95957.D4
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