Effect of Nitric Oxide on Anxiogenic-like Behavior Induced by Simvastatin Administration in Rats

Öz

Objective: Statins which inhibit cholesterol synthesis by inhibiting 3-hydroxy-3-methylglutaryl coenzyme A reductase enzymes are used widely for treatment of hyperlipidemia. In this study, altered anxiety levels by simvastatin use have been investigated to assess whether these effects are caused by changed nitric oxide levels.

Material and Methods: Seven groups of animals each consisting of 8 Sprague Dawley rats were tested by elevated plus maze and open field methods, to observe anxiety levels. The vehicle group was administered phosphate buffered saline alone while two of simvastatin groups were administered 20 or 40 mg/kg simvastatin and vehicle. Another two groups of animals were both administered 20 mg/kg simvastatin with vehicle. When one of these groups was additionally administered 10 mg/kg L-NAME, the other group was given 50 mg/kg L-NAME along with simvastatin. Remaining two groups were both given 10 and 50 mg/kg L-NAME doses along with 40 mg/kg simvastatin. All groups were tested in the 1st, 4th, 7th, 10th, 15th, 21st and 28th days of vehicle or drug administration. The research was analyzed using two way analyzes of variances.

Results: In the elevated plus maze test, simvastatin alone showed that the 40 mg / kg group significantly increased the closed arm time compared to the solvent group (p<0.05). Moreover, in the open field test, simvastatin 20 and 40 mg / kg groups significantly increased the margin duration compared to the solvent group (p<0.05). Doses of 10 and 50 mg/kg L-NAME abolished the effect created by simvastatin. The increase in closed arm time and margin time observed in the findings was evaluated as anxiogenic-like behavior.

Conclusion: The findings show that the anxiogenic-like behavior caused by the administration of simvastatin is prevented by the effect of L-NAME, thus the anxiogenic-like behavior caused by the administration of simvastatin is regulated by the nitric oxide mechanism.

Anahtar Kelimeler

• statins
• simvastatin
• anxiety
• nitric oxide
• elevated plus maze
• open field

SIÇANLARDA SİMVASTATİN UYGULAMASI İLE OLUŞAN ANKSİYOJENİK-BENZERİ DAVRANIŞTA NİTRİK OKSİTİN ETKİSİ

Öz

Amaç: 3-hidroksi-3-metil-glutaril-koenzim A redüktaz enzimini inhibe ederek kolesterol sentezini engelleyen statinler tüm dünyada hiperlipidemilerin tedavisinde yaygın olarak kullanılmaktadır. Statin ailesinin önemli bir üyesi olan simvastatinin, çeşitli çalışmalarda nitrik oksit miktarını arttırdığı gösterilmiştir. Bu çalışmada, simvastatin kullanımına bağlı olarak değişen anksiyete seviyesine, nitrik oksit düzeyinin etki edip etmediği araştırıldı.

Gereç ve Yöntemler: Sprague-Dawley cinsi sıçanlardan, her grupta 8 denek olmak üzere, 7 grupta deneklerin anksiyete düzeyinin gözlenmesi için yükseltilmiş artı labirent ve açık alan testi uygulandı. Çözücü grubuna fosfat tamponlu çözelti uygulanırken, yalnızca simvastatin alan gruba 20 ve 40 mg/kg dozda simvastatin uygulandı. Diğer iki gruptaki deneklere 20 mg/kg simvastatine ek olarak 10 mg/kg L-NAME, diğer gruba ise 50mg/kg L-NAME uygulandı. Kalan son iki gruba 40 mg/kg simvastatin ile 10 ve 50 mg/kg L-NAME dozları uygulandı. Çözücü ve ilaç uygulamasının 1, 4, 7, 10, 15, 21 ve 28. günlerinde tüm gruplar test edildi. Yükseltilmiş artı labirent kapalı kolda kalma süresi ve açık alan testi marjin zamanları ölçüldü. Sıçanlara uygulanan çeşitli simvastatin ve L-NAME dozlarının, yükseltilmiş artı labirent ve açık alan testine etkisi, iki yönlü varyans analizi kullanılarak değerlendirildi.

Bulgular: Yükseltilmiş artı labirent testinde sadece simvastatin 40 mg/kg grubunun çözücü grubuna göre kapalı kol kalma süresinin anlamlı olarak arttırdığını gösterdi (p<0.05). Ayrıca açık alan testinde simvastatin 20 ve 40 mg/kg grubu çözücü grubuna göre marjin süresini anlamlı olarak arttırdı (p<0.05). 10 ve 50 mg/kg L-NAME dozları simvastatinin yarattığı etkiyi ortadan kaldırdı. Bulgularda gözlenen kapalı kolda kalma zamanının ve marjin süresinin artışı anksiyojenik benzeri davranış olarak değerlendirilmiştir.

Sonuç: Bulgular simvastatin uygulaması ile oluşan anksiyojenik-benzeri davranışın L-NAME etkisiyle engellendiğini, dolayısıyla simvastatin uygulamasıyla oluşan anksiyojenik-benzeri davranışın nitrik oksit mekanizmasıyla düzenlendiğini göstermektedir.

Anahtar Kelimeler

• Statinler
• Simvastatin
• anksiyete
• Nitrik oksit
• yükseltilmiş artı labirent
• açık alan

Kaynakça

1. 1. Endo A, Kuroda M, Tsujita Y. ML-236A, ML-236B, and ML-236C, new inhibitors of cholesterogenesis produced by Penicillium citrinium. The Journal of antibiotics. 1976;29(12):1346-8.
2. 2. Fracassi A, Marangoni M, Rosso P, Pallottini V, Fioramonti M, Siteni S et al. Statins and the brain: more than lipid lowering agents? Current Neuropharmacology. 2019;17(1):59-83.
3. 3. Goldman JA, Fishman AB, Lee JE, Johnson RJ. The role of cholesterol-lowering agents in drug-induced rhabdomyolysis and polymyositis. Arthritis and rheumatism. 1989;32(3):358-9.
4. 4. McGirt MJ, Lynch JR, Parra A, Sheng H, Pearlstein RD, Laskowitz DT et al. Simvastatin increases endothelial nitric oxide synthase and ameliorates cerebral vasospasm resulting from subarachnoid hemorrhage. Stroke. 2002;33(12):2950-6.
5. 5. Laufs U, La Fata V, Plutzky J, Liao JK. Upregulation of endothelial nitric oxide synthase by HMG CoA reductase inhibitors. Circulation. 1998;97(12):1129-35.
6. 6. Nakata S, Tsutsui M, Shimokawa H, Yamashita T, Tanimoto A, Tasaki H et al. Statin treatment upregulates vascular neuronal nitric oxide synthase through Akt/NF-kappaB pathway. Arteriosclerosis, Thrombosis and Vascular Biology. 2007;27(1):92-8.
7. 7. Gao L, Wang W, Zucker IH. Simvastatin inhibits central sympathetic outflow in heart failure by a nitric-oxide synthase mechanism. The Journal of pharmacology and experimental therapeutics. 2008;326(1):278-85.
8. 8. Redondo-Morata L, Lea Sanford R, Andersen OS, Scheuring S. Effect of statins on the nanomechanical properties of supported lipid bilayers. Biophysical Journal. 2016;111(2):363-72.

9. 9. Pfrieger FW. Role of cholesterol in synapse formation and function. Biochimica et biophysica acta. 2003;1610(2):271-80.
10. 10. Vecka M, Tvrzicka E, Stankova B, Novak F, Novakova O, Zak A. Hypolipidemic drugs can change the composition of rat brain lipids. The Tohoku journal of experimental medicine. 2004;204(4):299-308.
11. 11. Thelen KM, Rentsch KM, Gutteck U, Heverin M, Olin M, Andersson U et al. Brain cholesterol synthesis in mice is affected by high dose of simvastatin but not of pravastatin. The Journal of Pharmacology and Experimental Therapeutics. 2006;316(3):1146-52.
12. 12. 12. Kureishi Y, Luo Z, Shiojima I, Bialik A, Fulton D, Lefer DJ et al. The HMG-CoA reductase inhibitor simvastatin activates the protein kinase Akt and promotes angiogenesis in normocholesterolemic animals. Nature Medicine. 2000;6(9):1004-10.
13. 13. Kesim M, Kadioglu M, Okuyan M, Muci E, Erkoseoglu I, Kalyoncu NI et al. The evaluation of analgesic effects of simvastatin, pravastatin and atorvastatin in hot plate test. European Review for Medical and Pharmacological Sciences. 2012;16(6):789-96.
14. 14. Sahin M, Ekinci M, Gedikli E, Baytan S. "Elevated Plus Maze on Based Computer Vision," 2007 IEEE 15th Signal Processing and Communications Applications, Eskisehir, Türkiye. 2007. pp.1-4.
15. 15. Schrader AJ, Taylor RM, Lowery-Gionta EG, Moore NLT. Repeated elevated plus maze trials as a measure for tracking within-subjects behavioral performance in rats (Rattus norvegicus). PloS one. 2018;13(11):e0207804.
16. 16. Binder E, Droste SK, Ohl F, Reul JM. Regular voluntary exercise reduces anxiety-related behaviour and impulsiveness in mice. Behavioural brain research. 2004;155(2):197-206.
17. 17. Baier CJ, Gallegos CE, Raisman-Vozari R, Minetti A. Behavioral impairments following repeated intranasal glyphosate-based herbicide administration in mice. Neurotoxicology and teratology. 2017;64:63-72. Doi:10.1016/j.ntt.2017.10.004 .
18. 18. Baytan SH, Alkanat M, Okuyan M, Ekinci M, Gedikli E, Ozeren M et al. Simvastatin impairs spatial memory in rats at a specific dose level. The Tohoku journal of experimental medicine. 2008;214(4):341-9.
19. 19. Baytan SH, Alkanat M, Ozeren M, Ekinci M, Akgun A. Fluvastatin alters psychomotor performance and daily activity but not the spatial memory in rats. The Tohoku journal of experimental medicine. 2006;209(4):311-20.
20. 20. Wang Q, Tang XN, Wang L, Yenari MA, Ying W, Goh BC et al. Effects of high dose of simvastatin on levels of dopamine and its reuptake in prefrontal cortex and striatum among SD rats. Neuroscience letters. 2006;408(3):189-93.
21. 21. Campos AC, Fogaca MV, Aguiar DC, Guimaraes FS. Animal models of anxiety disorders and stress. Revista Brasileira de Psiquiatria (Sao Paulo, Brazil : 1999). 2013;35 Suppl 2:S101-11.
22. 22. Cibickova L, Hyspler R, Micuda S, Cibicek N, Zivna H, Jun D et al. The influence of simvastatin, atorvastatin and high-cholesterol diet on acetylcholinesterase activity, amyloid beta and cholesterol synthesis in rat brain. Steroids. 2009;74(1):13-9.
23. 23. Wang Q, Wang PH, McLachlan C, Wong PT. Simvastatin reverses the downregulation of dopamine D1 and D2 receptor expression in the prefrontal cortex of 6-hydroxydopamine-induced Parkinsonian rats. Brain research. 2005;1045(1-2):229-33.
24. 24. Kuriyama K, Ohkuma S. Role of nitric oxide in central synaptic transmission: effects on neurotransmitter release. Japanese Journal of Pharmacology. 1995;69(1):1-8.
25. 25. Guimaraes FS, Beijamini V, Moreira FA, Aguiar DC, de Lucca AC. Role of nitric oxide in brain regions related to defensive reactions. Neuroscience and Biobehavioral Reviews. 2005;29(8):1313-22.
26. 26. Vincent SR, Kimura H. Histochemical mapping of nitric oxide synthase in the rat brain. Neuroscience. 1992;46(4):755-84.
27. 27. de Oliveira RW, Del Bel EA, Guimaraes FS. Behavioral and c-fos expression changes induced by nitric oxide donors microinjected into the dorsal periaqueductal gray. Brain Research Bulletin. 2000;51(6):457-64.
28. 28. Onstott D, Mayer B, Beitz AJ. Nitric oxide synthase immunoreactive neurons anatomically define a longitudinal dorsolateral column within the midbrain periaqueductal gray of the rat: analysis using laser confocal microscopy. Brain Research. 1993;610(2):317-24.
29. 29. Gulati K, Rai N, Ray A. Nitric oxide and anxiety. Vitamins and hormones. 2017;103:169-92. Doi:10.1016/bs.vh.2016.09.001
30. 30. Echeverry MB, Guimaraes FS, Del Bel EA. Acute and delayed restraint stress-induced changes in nitric oxide producing neurons in limbic regions. Neuroscience. 2004;125(4):981-93.
31. 31. Pshennikova MG, Bondarenko NA, Shimkovich MV. Nitric oxide as a factor of genetically determined resistance to stress damages and adaptive protection. Bulletin of experimental biology and medicine. 2001;132(5):1048-50.
32. 32. Volke V, Wegener G, Vasar E. Augmentation of the NO-cGMP cascade induces anxiogenic-like effect in mice. Journal of physiology and pharmacology: an official journal of the Polish Physiological Society. 2003;54(4):653-60.
33. 33. Slupski W, Trocha M, Rutkowska M. Pharmacodynamic and pharmacokinetic interactions between simvastatin and diazepam in rats. Pharmacological Reports. 2017;69(5):943-52.
34. 34. Ju A, Hanson CS, Banks E, Korda R, Craig JC, Usherwood T et al. Patient beliefs and attitudes to taking statins: systematic review of qualitative studies. Br J Gen Pract. 2018;68(671):e408-e419. Doi:10.3399/bjgp18X696365.
35. 35. Wang Q, Zengin A, Deng C, Li Y, Newell KA, Yang GY et al. High dose of simvastatin induces hyperlocomotive and anxiolytic-like activities: The association with the up-regulation of NMDA receptor binding in the rat brain. Experimental Neurology. 2009;216(1):132-8.
36. 36. Swanson CJ, Bures M, Johnson MP, Linden AM, Monn JA, Schoepp DD. Metabotropic glutamate receptors as novel targets for anxiety and stress disorders. Nature reviews Drug Discovery. 2005;4(2):131-44.
37. 37. Ferris P, Seward E, Dawson GR. Interactions between LY354740, a group II metabotropic agonist and the GABA(A)-benzodiazepine receptor complex in the rat elevated plus-maze. Journal of Psychopharmacology (Oxford, England). 2001;15(2):76-82.
38. 38. Monn JA, Valli MJ, Massey SM, Wright RA, Salhoff CR, Johnson BG et al. Design, synthesis, and pharmacological characterization of (+)-2-aminobicyclo[3.1.0]hexane-2,6-dicarboxylic acid (LY354740): a potent, selective, and orally active group 2 metabotropic glutamate receptor agonist possessing anticonvulsant and anxiolytic properties. Journal of Medicinal Chemistry. 1997;40(4):528-37.
39. 39. Ferraguti F. Metabotropic glutamate receptors as targets for novel anxiolytics. Curr Opin Pharmacol. 2018;38:37-42. Doi:10.1016/j.coph.2018.02.004.
40. 40. Cortese B, Phan KL. The role of glutamate in anxiety and related disorders. CNS spectrums. 2005;10:820-30.