Abstract
In a lipopolysaccharide (LPS)-induced rat model of sepsis (endotoxaemia), we previously demonstrated that pravastatin reduced microvascular inflammation via increased endothelial nitric oxide synthase III (NOSIII). This study aimed to determine whether atorvastatin, the most commonly used statin for lowering cholesterol, exerted beneficial pleiotropic effects via a similar mechanism. The mesenteric microcirculation of anaesthetised male Wistar rats (308 ± 63 g, n = 54) was prepared for fluorescent intravital microscopy. Over 4 h, animals received intravenous (i.v.) administration of either saline, LPS (150 μg kg−1 h−1) or LPS + atorvastatin (200 μg kg−1 s.c., 18 and 3 h before LPS), with/without the non-specific NOS inhibitor L-NG-Nitroarginine Methyl Ester (L-NAME) (10 μg kg−1 h−1) or NOSII-specific inhibitor 1400 W (20 μg kg−1 min−1). LPS decreased mean arterial blood pressure (MAP) (4 h, control 113 ± 20 mmHg; LPS 70 ± 23 mmHg), being reversed by atorvastatin (105 ± 3 mmHg) (p < 0.05). LPS also increased macromolecular leak measured after 100 mg kg−1 of i.v FITC-BSA (arbitrary grey level adjacent to venules), which again was attenuated by atorvastatin (control 1.9 ± 4.0; LPS 12.0 ± 2.4; LPS + atorvastatin 4.5 ± 2.2) (p < 0.05). Furthermore, immunohistochemistry identified that atorvastatin decreased LPS-induced upregulation of endothelial cell NOSII expression, but NOSIII was unchanged in all groups. Atorvastatin improved MAP and reduced microvascular inflammation during endotoxaemia, associated with a reduction of pro-inflammatory NOSII. This differs from previous studies, whereby pravastatin increased expression of NOSIII. Thus preoperative statins have beneficial anti-inflammatory effects during endotoxaemia, but careful consideration must be given to the specific statin being used.
Similar content being viewed by others
References
Andrew P, Deng Y, Kaufman S (2000) Fluid extravasation from spleen reduces blood volume in endotoxemia. Am J Physiol Regul Integr Comp Physiol 278:60–65
Bagnall NM, Dent PC, Walkowska A, Sadowski J, Johns EJ (2005) Nitric oxide inhibition and the impact on renal nerve-mediated antinatriuresis and antidiuresis in the anaesthetized rat. J Physiol 569:849–856
Birnbaum J, Lehmann C, Klotz E, Hein OV, Blume A, Jubin F, Polze N, Luther D, Spies CD (2008) Effects of N-acetylcysteine and tirilazad mesylate on intestinal functional capillary density, leukocyte adherence, mesenteric plasma extravasation and cytokine levels in experimental endotoxemia in rats. Clin Hemorheol Microcirc 39:99–111
Brookes ZL, Kaufman S (2005) Effects of atrial natriuretic peptide on the extrasplenic microvasculature and lymphatics in the rat in vivo. J Physiol 565:269–277
Brookes ZL, Brown NJ, Reilly CS (2002) Response of the rat cremaster microcirculation to hemorrhage in vivo: differential effects of intravenous anesthetic agents. Shock 18:542–548
Brookes ZL, McGown CC, Reilly CS (2009) Statins for all: the new premed? Br J Anaesth 103(1):99–107. doi:10.1093/bja/aep149
Calisto KL, Carvalho Bde M, Ropelle ER, Mittestainer FC, Camacho AC, Guadagnini D, Carvalheira JB, Saad MJ (2010) Atorvastatin improves survival in septic rats: effect on tissue inflammatory pathway and on insulin signaling. PLoS One 5(12):e14232
Clough GF, L’Esperance V, Turzyniecka M, Walter L, Chipperfield AJ, Gamble J, Krentz AJ, Byrne CD (2011) Functional dilator capacity is independently associated with insulin sensitivity and age in central obesity and is not improved by high dose statin treatment. Microcirculation 18:74–84
Dobesh PP, Klepser DG, McGuire TR, Morgan CW, Olsen KM (2009) Reduction in mortality associated with statin therapy in patients with severe sepsis. Pharmacotherapy 29:621–630. doi:10.1592/phco.29.6.621
Endres M, Laufs U, Huang Z, Nakamura T, Huang P, Moskowitz MA, Liao JK (1998) Stroke protection by 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase inhibitors (statins) unrelated to cholesterol lowering: new concepts for cardiovascular disease. Proc Natl Acad Sci U S A 95:8880–8885
Evans JM, Donnelly LA, Emslie-Smith AM, Alessi DR, Morris AD (2005) Metformin and reduced risk of cancer in diabetic patients. BMJ 330:1304–1305
Filep JG, Delalandre A, Beauchamp M (1997) Dual role for nitric oxide in the regulation of plasma volume and albumin escape during endotoxin shock in conscious rats. Circ Res 81:840–847
Fox SB, Leek RD, Weekes MP, Whitehouse RM, Gatter KC, Harris AL (1995) Quantitation and prognostic value of breast cancer angiogenesis: comparison of microvessel density, Chalkley count, and computer image analysis. J Pathol 177:275–283
Gardiner SM, Kemp PA, March JE, Bennett T (1995) Cardiac and regional haemodynamics, inducible nitric oxide synthase (NOS) activity, and the effects of NOS inhibitors in conscious, endotoxaemic rats. Br J Pharmacol 116:2005–2016
Garvey EP, Oplinger JA, Furfine ES, Kiff RJ, Laszlo F, Whittle BJ, Knowles RG (1997) 1400W is a slow, tight binding, and highly selective inhibitor of inducible nitric-oxide synthase in vitro and in vivo. J Biol Chem 272:4959–4963
Giusti-Paiva A, Martinez MR, Felix JV, da Rocha MJ, Carnio EC, Elias LL (2004) Simvastatin decreases nitric oxide overproduction and reverts the impaired vascular responsiveness induced by endotoxic shock in rats. Shock 21:271–275
Gong X, Ma Y, Ruan Y, Fu G, Wu S (2014) Long-term atorvastatin improves age-related endothelial dysfunction by ameliorating oxidative stress and normalizing eNOS/iNOS imbalance in rat aorta. Exp Gerontol 52:9–17
Grover R, Zaccardelli D, Colice G, Guntupalli K, Watson D, Vincent JL (1999) An open-label dose escalation study of the nitric oxide synthase inhibitor, N(G)-methyl-L-arginine hydrochloride (546C88), in patients with septic shock. Glaxo Wellcome International Septic Shock Study Group. Crit Care Med 27:913–922
Hickey MJ, Sharkey KA, Sihota EG (1997) Inducible nitric oxide synthase-deficient mice have enhanced leukocyte-endothelium interactions in endotoxemia. FASEB J 11:955–964
Hollenberg SM, Guglielmi M, Parrillo JE (2007) Discordance between microvascular permeability and leukocyte dynamics in septic inducible nitric oxide synthase deficient mice. Crit Care 11:125
Kandasamy K, Prawez S, Choudhury S, More AS, AHanger AA, Singh TU, Parida S, Mishra SK (2011) Atorvastatin prevents vascular hyporeactivity to norepinephrine in sepsis: role of nitric oxide and α-adrenoceptor mRNA expression. Shock 36(1):76–82. doi:10.1097/SHK.0b013e31821a4002
Kruger P, Bailey M, Bellomo R, Cooper DJ, Harward M, Higgins A, Howe B, Jones D, Joyce C, Kostner K, McNeil J, Nichol A, Roberts MS, Syres G, Venkatesh B, ANZ-STATInS Investigators–ANZICS Clinical Trials Group (2013) A multicenter randomized trial of atorvastatin therapy in intensive care patients with severe sepsis. Am J Respir Crit Care Med 187(7):743–750. doi:10.1164/rccm.201209-1718OC
Lahera V, Salom MG, Miranda-Guardiola F, Moncada S, Romero JC (1991) Effects of NG-nitro-L-arginine methyl ester on renal function and blood pressure. Renal Fluid Electrolyte Physiol 30:1033–F1037
Madonna R, Di Napoli P, Massaro M, Grilli A, Felaco M, De Caterina A, Tang D, De Caterina R, Geng YJ (2005) Simvastatin attenuates expression of cytokine-inducible nitric-oxide synthase in embryonic cardiac myoblasts. J Biol Chem 280:13503–13511
Matsuda N, Hayashi Y, Takahashi Y, Hattori Y (2006) Phosphorylation of endothelial nitric-oxide synthase is diminished in mesenteric arteries from septic rabbits depending on the altered phosphatidylinositol 3-kinase/Akt pathway: reversal effect of fluvastatin therapy. J Pharmacol Exp Ther 319:1348–1354
McGown CC, Brookes ZL (2007) Beneficial effects of statins on the microcirculation during sepsis: the role of nitric oxide. Br J Anaesth 98:163–175
McGown CC, Brown NJ, Hellewell PG, Reilly CS, Brookes ZL (2010) Beneficial microvascular and anti-inflammatory effects of pravastatin during sepsis involve NOSIII. Br J Anaesth 104:183–190. doi:10.1093/bja/aep361
Merx MW, Liehn EA, Janssens U, Lütticken R, Schrader J, Hanrath P, Weber C (2004) HMG-CoA reductase inhibitor simvastatin profoundly improves survival in a murine model of sepsis. Circulation 109(21):2560–2565
Merx MW, Liehn EA, Graf J, van de Sandt A, Schaltenbrand M, Schrader J, Hanrath P, Weber C (2005) Statin treatment after onset of sepsis in a murine model improves survival. Circulation 112:117–124
Miller FN, Joshua IG, Anderson GL (1982) Quantitation of vasodilator-induced macromolecular leakage by in vivo fluorescent microscopy. Microvasc Res 24:56–67
NICE (2008) NICE Implementation Uptake Report: statins for the prevention of cardiovascular events. National Institute for Health and Clinical Excellence. http://www.nice.org.uk/proxy/?sourceUrl=http%3a%2f%2fwww.nice.org.uk%2fmedia%2f919%2fC6%2fTA94UptakeReport.pdf. Accessed 23 Aug 2014
O’Neal HR Jr, Koyama T, Koehler EA, Siew E, Curtis BR, Fremonth RD, May AK, Bernard GR, Ware LB (2011) Prehospital statin and aspirin use and the prevalence of severe sepsis and acute lung injury/acute respiratory distress syndrome. Crit Care Med 39(6):1343–1350. doi:10.1097/CCM.0b013e3182120992
Patel JM, Snaith C, Thickett DR, Linhartova L, Melody T, Hawkey P, Barnett AH, Jones A, Hong T, Cooke MW, Perkins GD, Gao F (2012) Randomized double-blind placebo-controlled trial of 40 mg/day of atorvastatin in reducing the severity of sepsis in ward patients (ASEPSIS Trial). Crit Care 16(6):R231. doi:10.1186/cc11895
Scalia R, Stalker TJ (2002) Microcirculation as a target for the anti-inflammatory properties of Statins. Microcirculation 9:431–442
Simpson RJ Jr, Signorovitch J, Birnbaum H, Ivanova J, Connolly C, Kidolezi Y, Kuznik A (2009) Cardiovascular and economic outcomes after initiation of lipid-lowering therapy with atorvastatin vs simvastatin in an employed population. Mayo Clin Proc 84:1065–1072. doi:10.4065/mcp.2009.0298
Subramani J, Kathirvel K, Leo MD, Kuntamallappanavar G, Uttam Singh T, Mishra SK (2009) Atorvastatin restores the impaired vascular endothelium-dependent relaxations mediated by nitric oxide and endothelium-derived hyperpolarizing factors but not hypotension in sepsis. J Cardiovasc Pharmacol 54:526–534. doi:10.1097/FJC.0b013e3181bfafd6
Walther A, Weihrauch M, Schmidt W, Gebhard MM, Martin E, Schmidt H (2000) Leukocyte-independent plasma extravasation during endotoxemia. Crit Care Med 28:2943–2948
Wang L, Taneja R, Razavi HM, Law C, Gillis C, Mehta S (2012) Specific role of neutrophil inducible nitric oxide synthase in murine sepsis-induced lung injury in vivo. Shock 37(5):539–547. doi:10.1097/SHK.0b013e31824dcb5a
Xiao H, Qin X, Ping D, Zuo K (2013) Inhibition of Rho and Rac geranylgeranylation by atorvastatin is critical for preservation of endothelial junction integrity. PLoS One 8(3):e59233
Acknowledgments
We would also like to acknowledge assistance from Professor Tim Stephenson, Consultant Histopathologist, Sheffield Teaching Hospitals Trust, for his assistance and advice grading the IHC sections. Sectioning of slides was carried out by Yvonne Stephenson and Fiona Marrow (Histology Core Facility, University of Sheffield, Sheffield, UK).
Conflict of interest
None declared.
Funding
This work was supported by the Medical Research Council. Atorvastatin was a generous donation from Pfizer.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
McGown, C.C., Brookes, Z.L.S., Hellewell, P.G. et al. Atorvastatin reduces endotoxin-induced microvascular inflammation via NOSII. Naunyn-Schmiedeberg's Arch Pharmacol 388, 557–564 (2015). https://doi.org/10.1007/s00210-015-1100-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00210-015-1100-y