Aloe vera toxic effects: expression of inducible nitric oxide synthase (iNOS) in testis of Wistar rat

Document Type : Original Article

Authors

1 Department of Molecular Medicine, Faculty of Advanced Medical Technologies, Tehran University of Medical Sciences, Tehran, Iran

2 Medical Plant Research Center, Shahrekord University of Medical Sciences, Shahrekord, Iran

3 Cellular and Molecular Research Center, Shahrekord University of Medical Sciences, Shahrekord, Iran

4 Department of Genetics, Faculty of Basic Sciences, University of Shahrekord, Shahrekord, Iran

Abstract

Objective(s): Nitric oxide (NO), a product of inducible nitric oxide synthase (iNOS), contributes in germ cell apoptosis. This study was aimed to evaluate the effects of Aloe vera gel (AVG) on male Wistar rat reproductive organ, serum NO level, and expression of iNOS gene in leydig cells.
Materials and Methods:Adult male Wistar rats (n=36) were used for experiments in three groups. The experimental groups were orally administered with the AVG extract solution once-daily as follow: 150 mg.kg-1; group A, 300 mg.kg-1; group B, and only normal saline; group C (control group). They were mated with untreated females and the reproductive and chemical parameters were assessed for each group, including semen quality, serum testosterone, sperm fertility, gonad and body weight, serum NO concentration (by the Griess method), and iNOS gene expression (using RT-PCR).
Results: The testes weight, serum testosterone, as well as sperm count and fertility of the AVG treated groups were significantly reduced when compared to the control (P<0.001). Concentration of serum NO was significantly increased (37.1±4.63 µM) in the administrated group with higher AVG concentration, compared to the control group (P<0.001; 10.19±0.87 µM); however, iNOS mRNA expression was increased in the treated animals (P<0.001).
Conclusion:iNOS may play a functional role in spermatogenesis via apoptosis, reducing sperm count, but further studies are needed to illustrate the mechanisms by which AVG exerts its negative effects on spermatogenesis and sperm quality.

Keywords

References

 
1.Weissman BA, Sottas CM, Zhou P, Iadecola C, Hardy MP. Testosterone production in mice lacking inducible nitric oxide synthase expression is sensitive to restraint stress. Am J Physiol Endocrinol 2007; 292:615-620.
2.Shirzad H, Nasri H. Toxicity and safety of medicinal plants. J Herb Med Pharmacol 2013; 2:21-22.
3.Rafieian-kopaei M, Shahinfard N, Rouhi-Boroujeni H, Gharipour M, Darvishzadeh-Boroujeni P. Effects of Ferulago angulata extract on serum lipids and lipid peroxidation. J Evid Based Complementary Altern Med 2014; 2014:4.
4.Bahmani M, Zargaran A, Rafieian-Kopaei M, Saki M. Ethnobotanical study of medicinal plants used in the management of diabetes mellitus in the Urmia, Northwest Iran. Asian Pac J Trop Med 2014; 7: 348-354. 
5.Haniadka R, Kamble PS, Azmidha A, Mane PP, Geevarughese NM, Palatty PL, et al. Review on the use of Aloe vera (Aloe) in dermatology. Nutr Health 2013; 125-133.
6.Yagi A. Possible prophylactic eEfficacy of Aloe vera for γ-ray radiation-induced damages. J Gastroen Hepatol  2013; 2.
7.Suganya S, Venugopal J, Mary SA, Ramakrishna S, Lakshmi B, Dev VG. Aloe vera incorporated biomimetic nanofibrous scaffold: a regenerative approach for skin tissue engineering. Iran Polym J 2014; 23:237-248.
8.Baradaran A, Nasri H, Nematbakhsh M, Rafieian-Kopaei M. Antioxidant activity and preventive effect of aqueous leaf extract of Aloe vera on gentamicin-induced nephrotoxicity in male Wistar rats. Clin Ter 2014; 165: 7-11.
9.Delfan B, Bahmani M, Eftekhari Z, Jelodari M, Saki K, Mohammadi T. Effective herbs on the wound and skin disorders: a ethnobotanical study in Lorestan province, west of Iran. Asian Pac J Trop Dis 2014; 4:938-942.
10. Grace OM, Dzajic A, Jäger AK, Nyberg NT, Önder A, Rønsted N. Monosaccharide analysis of succulent leaf tissue in Aloe. Phytochemistry 2013; 93:79-87.
11. Ray A, Aswatha SM. An analysis of the influence of growth periods on physical appearance, and acemannan and elemental distribution of Aloe vera gel. Ind Crops Prod 2013; 48:36-42.
12. Djeraba A, Quere P. In vivo macrophage activation in chickens with Acemannan, a complex carbohydrate extracted from Aloe vera. Int J Immunopharmacol 2000; 22:365-372.
13. Budai MM, Varga A, Milesz S, Tőzsér J, Benkő S. Aloe vera downregulates LPS-induced inflammatory cytokine production and expression of NLRP3 inflammasome in human macrophages. Mol Immunol 2013; 56:471-479.
14. Klein-Wieringa IR, Andersen NS, Kwekkeboom JC,  Giera M, de Lange-Brokaar BJ, van Osch GJ, et al. Adipocytes modulate the phenotype of human macrophages through secreted lipids. J Immunol 2013; 191:1356-1363.
15. YanL, Li Z. Mannose receptor ligands regulate the gene expression of Toll-like receptors in chicken monocytes. J Poult Sci 2013; 50:388-395.
16. Pitsikas N. The role of nitric oxide in the object recognition memory. Behav Brain Res 2015; 285: 200–207.
17. Jarazo-Dietrich S, Jacobo P, Pérez C, Guazzone V, Lustig L, Theas M. Up regulation of nitric oxide synthase–nitric oxide system in the testis of rats undergoing autoimmune orchitis. Immunobiology 2012; 217: 778-787.
18. Andric SA, Janjic MM, Stojkov NJ, Kostic TS. Testosterone-induced modulation of nitric oxide-cGMP signaling pathway and androgenesis in the rat Leydig cells. Biol Reprod 2010; 83:434-442.
19. Ikegami H, Kajikawa S, Ito K, Nii A, Okamiya H, Nakayama H, et al. Immunohistochemical study on inducible type of nitric oxide (iNOS), basic fibroblast growth factor (bFGF) and tumor growth factor-β1 (TGF-β1) in arteritis induced in rats by fenoldopam and theophylline, vasodilators. Exp Toxicol Pathol 2002; 54:1-7.
20. Lal B, Dubey N. Existence of a nitric oxide synthase/nitric oxide system in fish testis and its role in modulation of androgenesis. Fish Physiol Biochem 2013; 39:65-69.
21. Auharek S, Lara N, Avelar G, Sharpe R, França L. Effects of inducible nitric oxide synthase (iNOS) deficiency in mice on Sertoli cell proliferation and perinatal testis development. Int J Androl 2012; 35: 741-751.
22. Yu W, Zheng H, Lin W, Tajima A, Zhang Y, Zhang X, et al. Estrogen promotes Leydig cell engulfment by macrophages in male infertility. J  Clin Invest 2014; 124:2709–27210.
23. Valenti S, Cuttica CM, Fazzuoli L, Giordano G, Giusti M. Biphasic effect of nitric oxide on testosterone and cyclic GMP production by purified rat Leydig cells cultured in vitro. Int J Androl 1999; 22:336-41.
24. Rajasekaran S, Sivagnanam K, Subramanian S. Antioxidant effect of Aloe vera gel extract in streptozotocin-induced diabetes in rats. Pharmacol Report 2005; 57: 90-96.
25. Rabiei Z, Rafieian-kopaei M, Heidarian E, Saghaei E, Mokhtari S. Effects of zizyphus jujube extract on memory and learning impairment induced by bilateral electric lesions of the nucleus basalis of meynert in rat. Neurochem Res 2014; 39: 353-360.
26. Rabiei Z, M Rafieian-Kopaei. Neuroprotective effect of pretreatment with Lavandula officinalis ethanolic extract on blood-brain barrier permeability in a rat stroke model. Asian Pac J Trop Med 2014; 7: 421-426.
27. Rabiei Z, Rafieian-kopaei M,  Mokhtari  S, Shahrani M. Effect of dietary ethanolic extract of lavandula officinalis on serum lipids profile in pats. Iran J Pharm Res 2014; 13:1295–1301.
28. Rajasekaran S, Sivagnanam K, Ravi K, Subramanian S. Hypoglycemic effect of Aloe vera gel on streptozotocin-induced diabetes in experimental rats. J Med Food 2004; 7:61-66.
29. Rajasekaran S, Sivagnanam K, Ravi K, Subramanian S. Beneficial effects of Aloe vera leaf gel extract on lipid profile status in rats with streptozotocin diabetes. Clin Exp Pharmacol Physiol 2006; 33:232-237.
30. Lucio RA, Tlachi-López JL, Eguibar JR, Ågmo A. Sperm count and sperm motility decrease in old rats. Physiol Behav 2013; 110–111:73-79.
31. Navarro-Gonzálvez JA, García-Benayas C, Arenas J. Semiautomated measurement of nitrate in biological fluids. Clin Chem 1998; 44:679-681.
32. Dimmeler S, Zeiher A. Nitric Oxide and Apoptosis: Another paradigm for the double-edged role of nitric Oxide. Nitric Oxide 1997; 2:75–281.
33. Cohen O, Ish-Shalom E, Kfir-Erenfeld S, Herr I, Yefenof E. Nitric oxide and glucocorticoids synergize in inducing apoptosis of CD4+8+thymocytes: implications for ‘Death by Neglect’ and T-cell function. Int Immunol 2012; 24:783-791.
34. F, Ni B, Liu M, Feng Z, Xiong Q, Xiao S, Shao G. Mycoplasma hyopneumoniae-derived lipid-associated membrane proteins induce apoptosis in porcine alveolar macrophage via increasing nitric oxide production, oxidative stress, and caspase-3 activation. Vet Immunopathol 2013; 55–161.
35. Saini A, Shenoy G, Rath S, Bal V, George A. Inducible nitric oxide synthase is a major intermediate in signaling pathways for the survival of plasma cells. Nat Immunol 2014; 15:275–228.
36. Lue Y, Sinha Hikim AP, Wang C, Leung A, Swerdloff  RS. Functional role of inducible nitric oxide synthase in the induction of male germ cell apoptosis, regulation of sperm number, and determination of testes size: evidence from null mutant mice. J Endocrinol 2003; 144:3092-3100.
37. Chen D,  Zheng W,  Lin A, Uyhazi K,  Zhao H,  Lin H. Pumilio 1 suppresses multiple activators of p53 to safeguard spermatogenesis. Curr Biol 2012; 22:420-425.
38. Broniowska KA, Diers AR, Corbett JA, Hogg N. Effect of nitric oxide on naphthoquinone toxicity in endothelial cells: role of bioenergetic dysfunction and poly(ADP-ribose) polymerase activation. Biochemistry 2013; 52:4364–4372.
39. Nee Pathak ND, Lal B. Paracrine role of macrophage produced-nitric oxide (NO) in Leydig cell steroidogenesis in a teleost, Clarias batrachus: Impact of gonadotropin, growth hormone and insulin on NO production by testicular macrophages. Gen Comp Endocrinol 2009; 160:12-18.
40. Karimi Jashni HH, Najmadini N, Hooshm F. Effect of alcoholic extract of Aloe vera plant on serum testosterone and gonadotropin levels in rats. J Jahrom Univ Med Sci 2012; 10:1-8.
41. Skarzynski DJ, Kobayashi S, Okuda K. Influence of nitric oxide and noradrenaline on prostaglandin F2α-induced oxytocin secretion and intracellular calcium mobilization in cultured bovine luteal cells. Biol Reprod 2000; 63:1000-1005.
42. Mondillo C, Pagotto RM, Piotrkowski B, Reche CG, Patrignani ZJ, Cymeryng CB, et al. Involvement of nitric oxide synthase in the mechanism of histamine-induced inhibition of Leydig cell steroidogenesis via histamine receptor subtypes in Sprague-Dawley rats. Biol Reprod 2009; 80:144-152.
43. Nee Pathak ND, Lal B. Nitric oxide: An autocrine regulator of Leydig cell steroidogenesis in the Asian catfish, Clarias batrachus. Gen Comp Endocrinol 2008; 158:161-167.
44. Elmore S. Apoptosis: a review of programmed cell death. Toxicol Pathol 2007; 35:495-516.
45. Brookes PS, Levonen AL, Shiva S, Sarti S, Darley-Usmar VM. Mitochondria: regulators of signal   transduction by reactive oxygen and nitrogen species. Free Radic Biol Med 2002; 33:755-764.
46. Sinha-Hikim I, Braga M, Shen R, Hikim APS. Involvement of c-Jun NH2-terminal kinase and nitric oxide-mediated mitochondria-dependent intrinsic pathway signaling in cardiotoxin-induced muscle cell death: role of testosterone. Apoptosis 2007; 12:1965-1978.
47.Ghatan S, Larner S, Kinoshita Y, Hetman M, Patel L, Xia Z, et al. P38 MAP kinase mediates BAX translocation in nitric oxide–induced apoptosis in neurons. J Cell Biol 2000; 150:335-348.
48. Hikim APS, Lue Y, Diaz-Romero M, Yen PH, Wang C, Swerdloff  RS. Deciphering the pathways of germ cell apoptosis in the testis. J Steroid Biochem Mol Biol 2003; 85:175-182.
49. Tripathi R, Mishra DP,Shaha C. Male germ cell development: turning on the apoptotic pathways. J Reprod Immunol 2009; 83:31-35.
50. Yamamoto CM, Sinha Hikim AP, Huynh PN, Shapiro B, Lue YH, Salameh WA, et al.  Redistribution of Bax is an early step in an apoptotic pathway leading to germ cell death in rats, triggered by mild testicular hyperthermia. Biol Reprod 2000; 63:1683–1690.
Iranian Journal of Basic Medical Sciences
Volume 18, Issue 10 - Serial Number 10
October 2015
Pages 967-973

Files

Share

How to cite

Statistics