NON â€“ RECEPTOR MEDIATED ANXIOLYSIS BY AMINO GUANIDINE: A NOVEL ACTIVITY AND SYNERGY WITH DIAZEPAM UNDER STRESSED CONDITIONS
Objective: A novel non-receptor mediated anxiolytic activity of amino guanidine (AG; 50 mg/kg, i.p.) was investigated and its possible interaction
with diazepam (DZP) (2 mg/kg, i.p.) was explored in mice.
Methods: Elevated plus maze and light/dark box were used to measure anxiety in Swiss albino mice. Plasma nitrite levels were estimated using Griess
reagent to observe the biochemical regulation of mice behavior.
Results: AG (50 mg/kg, i.p.) produced a significant antianxiety-like activity and significantly attenuated the plasma nitrite levels in stressed mice.
On the other hand, AG (50 mg/kg, i.p.) neither produced a significant antianxiety-like activity nor significantly altered the plasma nitrite levels in
unstressed mice. DZP (2 mg/kg, i.p.) produced a significant antianxiety-like activity in unstressed mice but not in stressed mice. DZP (2 mg/kg, i.p.)
did not significantly affect plasma nitrite levels in unstressed or stressed mice. AG and DZP, administered together produced a significant antianxietylike
activity in unstressed and stressed mice. However, combination of AG and DZP significantly attenuated the plasma nitrite levels in stressed mice
but not in unstressed mice.
Conclusion: These findings suggest a usefulness of a non-GABAergic mechanism of a potential anti-anxiety chemical that may avoid receptor-linked
undesired effects as in case of DZP and may be used with DZP under stressed conditions to serve the reduction of dosage of DZP.
Keywords: Amino guanidine, Anxiety, Diazepam, Stress.
2. Tsuchiya T, Kishimoto J, Koyama J, Ozawa T. Modulatory effect of L-NAME, a specific nitric oxide synthase (NOS) inhibitor, on stress-induced changes in plasma adrenocorticotropic hormone (ACTH) and corticosterone levels in rats: Physiological significance of stress-induced NOS activation in hypothalamic-pituitary-adrenal axis. Brain Res 1997;776(1-2):68-74.
3. Madrigal JL, Hurtado O, Moro MA, Lizasoain I, Lorenzo P, Castrillo A, et al. The increase in TNF-alpha levels is implicated in NF-kappaB activation and inducible nitric oxide synthase expression in brain cortex after immobilization stress. Neuropsychopharmacology 2002;26(2):155-63.
4. Sevgi S, Ozek M, Eroglu L. L-NAME prevents anxiety-like and depression-like behavior in rats exposed to restraint stress. Methods Find Exp Clin Pharmacol 2006;28(2):95-9.
5. Hata T, Nishikawa H, Itoh E, Funakami Y. Anxiety-like behavior in elevated plus-maze tests in repeatedly cold-stressed mice. Jpn J Pharmacol 2001;85(2):189-96.6. Albonetti ME, Farabollini F. Behavioural responses to single and repeated restraint in male and female rats. Behav Processes 1992;28(1â€‘2):97-109.
7. Belzung C, Griebel G. Measuring normal and pathological anxiety-like behaviour in mice: A review. Behav Brain Res 2001;125(1-2):141-9.
8. Kim CK, Rivier CL. Nitric oxide and carbon monoxide have a stimulatory role in the hypothalamic-pituitary-adrenal response to physico-emotional stressors in rats. Endocrinology 2000;141(6):2244â€‘53.
9. Harada S, Imaki T, Chikada N, Naruse M, Demura H. Distinct distribution and time-course changes in neuronal nitric oxide synthase and inducible NOS in the paraventricular nucleus following lipopolysaccharide injection. Brain Res 1999;821(2):322-32.
10. Heinrichs SC, Pich EM, Miczek KA, Britton KT, Koob GF. Corticotropin-releasing factor antagonist reduces emotionality in socially defeated rats via direct neurotropic action. Brain Res 1992;581(2):190-7.
11. Quock RM, Nguyen E. Possible involvement of nitric oxide in chlordiazepoxide-induced anxiolysis in mice. Life Sci 1992;51(25):PL255-60.
12. MonzÃ³n ME, Varas MM, De Barioglio SR. Anxiogenesis induced by nitric oxide synthase inhibition and anxiolytic effect of melanin-concentrating hormone (MCH) in rat brain. Peptides 2001;22(7):1043â€‘7.
13. Roohbakhsh A, Moghaddam AH, Massoudi R, Zarrindast MR. Role of dorsal hippocampal cannabinoid receptors and nitric oxide in anxiety like behaviours in rats using the elevated plus-maze test. Clin Exp Pharmacol Physiol 2007;34(3):223-9.
14. Volke V, Soosaar A, KÃµks S, Bourin M, MÃ¤nnistÃ¶ PT, Vasar E. 7-Nitroindazole, a nitric oxide synthase inhibitor, has anxiolytic-like properties in exploratory models of anxiety. Psychopharmacology (Berl) 1997;131(4):399-405.
15. Yildiz F, Ulak G, Erden BF, Gacar N. Anxiolytic-like effects of 7-nitroindazole in the rat plus-maze test. Pharmacol Biochem Behav 2000;65(2):199-202.
16. Forestiero D, Manfrim CM, GuimarÃ£es FS, de Oliveira RM. Anxiolytic-like effects induced by nitric oxide synthase inhibitors microinjected into the medial amygdala of rats. Psychopharmacology (Berl) 2006;184(2):166-72.
17. SpolidÃ³rio PC, Echeverry MB, Iyomasa M, GuimarÃ£es FS, Del Bel EA. Anxiolytic effects induced by inhibition of the nitric oxide-cGMP pathway in the rat dorsal hippocampus. Psychopharmacology (Berl) 2007;195(2):183-92.
18. Stuehr DJ, Marletta MA. Induction of nitrite/nitrate synthesis in murine macrophages by BCG infection, lymphokines, or interferon-gamma. J Immunol 1987;139(2):518-25.
19. FÃ¶rstermann U, Kleinert H. Nitric oxide synthase: Expression and expressional control of the three isoforms. Naunyn Schmiedebergs Arch Pharmacol 1995;352(4):351-64.
20. Olivenza R, Moro MA, Lizasoain I, Lorenzo P, FernÃ¡ndez AP, Rodrigo J, et al. Chronic stress induces the expression of inducible nitric oxide synthase in rat brain cortex. J Neurochem 2000;74(2):785â€‘91.
21. Madrigal JL, Moro MA, Lizasoain I, Lorenzo P, Castrillo A, BoscÃ¡ L, et al. Inducible nitric oxide synthase expression in brain cortex after acute restraint stress is regulated by nuclear factor kappaB-mediated mechanisms. J Neurochem 2001;76(2):532-8.
22. Sapolsky RM, Uno H, Rebert CS, Finch CE. Hippocampal damage associated with prolonged glucocorticoid exposure in primates. J Neurosci 1990;10(9):2897-902.
23. Sheline YI, Wang PW, Gado MH, Csernansky JG, Vannier MW. Hippocampal atrophy in recurrent major depression. Proc Natl Acad Sci U S A 1996;93(9):3908-13.
24. Southan GJ, SzabÃ³ C. Selective pharmacological inhibition of distinct nitric oxide synthase isoforms. Biochem Pharmacol 1996;51(4):383â€‘94.
25. Klandorf H, Zhou Q, Sams AR. Inhibition by aminoguanidine of glucose-derived collagen cross-linking in skeletal muscle of broiler breeder hens. Poult Sci 1996;75(3):432-7.
26. Lee FY, Wang SS, Tsai YT, Lin HJ, Lin HC, Chu CJ, et al. Aminoguanidine corrects hyperdynamic circulation without ameliorating portal hypertension and portal hypertensive gastropathy in anesthetized portal hypertensive rats. J Hepatol 1997;26(3):687-93.
27. Gilhotra N, Dhingra D. Involvement of NO-cGMP pathway in anti-anxiety effect of aminoguanidine in stressed mice. Prog Neuropsychopharmacol Biol Psychiatry 2009;33(8):1502-7.
28. Schwartz TL, Nihalani N, Simionescu M, Hopkins G. History repeats itself: Pharmacodynamic trends in the treatment of anxiety disorders. Curr Pharm Des 2005;11(2):255-63.
29. Elie R, Lamontagne Y. Alprazolam and diazepam in the treatment of generalized anxiety. J Clin Psychopharmacol 1984;4(3):125-9.
30. Saddock BJ, Saddock VA. Kaplan & Saddockâ€™s Pocket Handbook of Psychiatric Drug Treatment. 3rd ed. USA: Lippincoat, Williams, & Wilkins; 2001.
31. Talarek S, Listos J, Fidecka S. Role of nitric oxide in the development of tolerance to diazepam-induced motor impairment in mice. Pharmacol Rep 2008;60(4):475-82.
32. Talarek S, Orzelska J, Listos J, Fidecka S. Effects of sildenafil treatment on the development of tolerance to diazepam-induced motor impairment and sedation in mice. Pharmacol Rep 2010;62(4):627-34.
33. Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR. Analysis of nitrate, nitrite, and [15N] nitrate in biological fluids. Anal Biochem 1982;126(1):131-8.
34. Bartos J, Pesez M. Colorimetric and fluorimetric determination of steroids. Pure Appl Chem 1979;51:2157-69.
35. Calatayud F, Belzung C, Aubert A. Ethological validation and the assessment of anxiety-like behaviours: Methodological comparison of classical analyses and structural approaches. Behav Processes 2004;67(2):195-206.
36. Harvey BH, Oosthuizen F, Brand L, Wegener G, Stein DJ. Stress-restress evokes sustained iNOS activity and altered GABA levels and NMDA receptors in rat hippocampus. Psychopharmacology (Berl) 2004;175(4):494-502.
37. Gilhotra N, Dhingra D. GABAergic and nitriergic modulation by curcumin for its antianxiety-like activity in mice. Brain Res 2010;1352:167-75.
38. Volke V, Soosaar A, KÃµks S, Vasar E, MÃ¤nnistÃ¶ PT. L-Arginine abolishes the anxiolytic-like effect of diazepam in the elevated plus-maze test in rats. Eur J Pharmacol 1998;351(3):287-90.
39. Goyal R, Anil K. Protective effect of alprazolam in acute immobilization stress-induced certain behavioral and biochemical alterations in mice. Pharmacol Rep 2007;59(2):284-90.
40. Kumari B, Kumar A, Dhir A. Protective effect of non-selective and selective COX-2-inhibitors in acute immobilization stress-induced behavioral and biochemical alterations. Pharmacol Rep 2007;59(6):699â€‘707.
41. Gilhotra N, Jain H, Dhingra D. Differential effects of nitric oxide synthase inhibitors on anxiety in unstressed and stressed mice. Indian J Exp Biol 2010;48(4):365-72.
42. Bhattacharya SK, Bhattachatyya D. Effect of restraint stress on rat brain serotonin. J Biosci 1982;4:269-74.
43. Korte SM, De Boer SF. A robust animal model of state anxiety: Fear-potentiated behaviour in the elevated plus-maze. Eur J Pharmacol 2003;463(1-3):163-75.
44. Bremner JD. Animal models for the neurobiology of trauma. PTSD Res Quart 1991;2:1.
45. Sharma V, Gilhotra R, Dhingra D, Gilhotra N. Possible underlying influence of p38MAPK and NF-ÎºB in the diminished anti-anxiety effect of diazepam in stressed mice. J Pharmacol Sci 2011;116(3):257â€‘63.
46. Concas A, Mele S, Biggio G. Foot shock stress decreases chloride efflux from rat brain synaptoneurosomes. Eur J Pharmacol 1987;135(3):423â€‘7.
47. Drugan RC, Morrow AL, Weizman R, Weizman A, Deutsch SI, Crawley JN, et al. Stress-induced behavioral depression in the rat is associated with a decrease in GABA receptor-mediated chloride ion flux and brain benzodiazepine receptor occupancy. Brain Res 1989;487(1):45-51.
48. Weizman A, Bidder M, Fares F, Gavish M. Food deprivation modulates gamma-aminobutyric acid receptors and peripheral benzodiazepine binding sites in rats. Brain Res 1990;535(1):96-100.
49. Gilhotra N, Dhingra D. Thymoquinone produced antianxiety-like effects in mice through modulation of GABA and NO levels. Pharmacol Rep 2011;63(3):660-9.
50. Krsiak M. Timid singly-housed mice: Their value in prediction of psychotropic activity of drugs. Br J Pharmacol 1975;55:141-50.
51. Kurt M, Bilge SS, Aksoz E, Kukula O, Celik S, Kesim Y. Effect of sildenafil on anxiety in the plus-maze test in mice. Pol J Pharmacol 2004;56(3):353-7.
52. Faria MS, MuscarÃ¡ MN, Moreno JÃºnior H, Teixeira SA, Dias HB, De Oliveira B, et al. Acute inhibition of nitric oxide synthesis induces anxiolysis in the plus maze test. Eur J Pharmacol 1997;323(1):37-43.
53. Tsuji M, Takeda H, Matsumiya T. Different effects of 5-HT1A receptor agonists and benzodiazepine anxiolytics on the emotional state of naive and stressed mice: A study using the hole-board test. Psychopharmacology (Berl) 2000;152(2):157-66.
54. Cowen PJ, Nutt DJ. Abstinence symptoms after withdrawal of tranquillising drugs: Is there a common neurochemical mechanism? Lancet 1982;2(8294):360-2.
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