Skip to main content
SpringerLink
Log in

Determination of histological, immunohistochemical and biochemical effects of acute and chronic grayanotoxin III administration in different doses in rats

  • Research Article
  • Published:
Environmental Science and Pollution Research Aims and scope Submit manuscript

Abstract

Grayanotoxin (GTX)-III is a Na-channel neurotoxin. Grayanotoxins can be found in the nectar, pollen, and other plant parts of the Rhododendron genus plants from the Ericaceae family. It is widely believed that honey produced from these plants, which are concentrated in the Black Sea region, is traditionally characterized as enhancing sexual performance. It is thought that the effective factor is dose for this compound, which has both beneficial and toxic effects reported. Therefore, it is aimed to evaluate the histological, immunohistochemical, and biochemical effects of acute and chronic impact of GTX-III in different doses on testes tissue in this study. For this purpose, 100 Sprague-Dawley male rats were divided into 5 separate groups for acute and chronic research. While dose groups were (control, 0.1, 0.2, 0.4, ve 0.8 μg/kg/bw) for experimental groups, a single dose (i.p.) was administered for acute impact whereas the same doses were administered daily for 3 weeks to assess chronic effect. At the end of the experiment, Johnsen testicular biopsy scoring was performed on testicular tissue samples, seminiferous tubule diameters were measured, and apoptotic cells were evaluated by TUNEL method. Testosterone, LH, and FSH levels were measured by ELISA method in serum and tissue specimens. It was found that Johnsen score of acute doses was significantly lower than the control group, and the diameter of the seminiferous tubules decreased significantly in acute and chronic dose-administered groups compared to the control. Hemorrhage, epithelial shedding, irregularity in seminiferous epithelium, and vacuolization were observed in acute and chronic dose-administered groups, and increase in apoptotic cells was determined. Hormone levels varied depending on the dose. In conclusion, it was found that dose-dependent acute and chronic effects of GTX-III are different, and this factor should be taken into account in studies to be carried out due to the adverse effects of high doses.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Ahlbom E, Prins GS, Ceccatelli S (2001) Testosterone protects cerebellar granule cells from oxidative stress-induced cell death through a receptor mediated mechanism. Brain Res 892(2):255–262

    Article  CAS  Google Scholar 

  • Ascioglu C, Ozesmi C, Dogan P, Ozturk F (2000) Effects of acute grayantoxin-I administration on hepatic and renal functions in rats. Turkish J Med Sci 30:23–27

    CAS  Google Scholar 

  • Aydilek N, Aksakal M, Karakilçik AZ (2004) Effects of testosterone and vitamin E on the antioxidant system in rabbit testis. Andrologia 36(5):277–281

    Article  CAS  Google Scholar 

  • Chainy GB, Samantaray S, Samanta L (1997) Testosterone-induced changes in testicular antioxidant system. Andrologia 29(6):343–349

    Article  CAS  Google Scholar 

  • Constantine GH, Sheth K, Catalfomo P (1967) Grayanotoxin I. Occurrence in additional Ericaceae species. J Pharm Sci 56:1518–1519

    Article  CAS  Google Scholar 

  • Demircan A, Keleş A, Bildik F, Aygencel G, Doğan NÖ, Gómez HF (2009) Mad honey sex: therapeutic misadventures from an ancient biological weapon. Ann Emerg Med 54(6):824–829

    Article  Google Scholar 

  • Den Boer PJ, Poot M, Verkerk A (1990) Glutathione-dependent defence mechanisms in isolated round spermatids from the rat. Int J Androl 13(1):26–38

    Article  Google Scholar 

  • Fallahzadeh AR, Rezaei Z, Rahimi HR, Barmak MJ, Sadeghi H, Mehrabi S, Rabani SM, Kashani IR, Barati V, Mahmoudi R (2017) Evaluation of the effect of Pentoxifylline on cisplatin-induced testicular toxicity in rats. Toxicol Res 33:255–263

    Article  CAS  Google Scholar 

  • Finkel T, Holbrook NJ (2000) Oxidants, oxidative stress and the biology of ageing. Nature 408(6809):239–247

    Article  CAS  Google Scholar 

  • Ghasemnezhad R, Mohammadghasemi F, Faghani M, Bahadori MH (2015) Oxytocin can decrease germ cells apoptotic index in testis under acute ischemia reperfusion in a rat model. Iran J Reprod Med 13:283–290

    Google Scholar 

  • Gholami M, Abbaszadeh A, Khanipour Khayat Z (2018) Honey improves spermatogenesis and hormone secretion in testicular ischaemia–reperfusion-induced injury in rats. Andrologia 50:e12804

    Article  CAS  Google Scholar 

  • Gil D, Leboucher G, Lacroix A, Cue R, Kreutzer M (2004) Female canaries produce eggs with greater amounts of testosterone when exposed to preferredmale song. Horm Behav 45(1):64–70

    Article  CAS  Google Scholar 

  • Gunduz A, Turedi S, Uzun H, Topbas M (2006) Mad honey poisoning. Am J Emerg Med 24:595–598

    Article  Google Scholar 

  • Gunduz A, Turedi S, Russell RM, Ayaz F (2008) Clinical review of grayanotoxin/mad honey poisoning past and present. Clin Toxicol (Phila) 46:437–442

    Article  CAS  Google Scholar 

  • Gunduz A, Merice ES, Baydin A et al (2009) Does mad honey poisoning require hospital admission? Amer J Emerg Med 27(4):424–427

    Article  Google Scholar 

  • Gunduz A, Eraydin I, Turkmen S, Kalkan OF, Turedi S, Eryigit U, Ayar A (2014) Analgesic effects of mad honey (grayanotoxin) in mice models of acute pain and painful diabetic neuropathy. Hum Exp Toxicol 33:130–135

    Article  CAS  Google Scholar 

  • Halliwell B, Gutteridge JM (1985) The importance of free radicals and catalytic metal ions in human diseases. Mol Asp Med 8(2):89–193

    Article  CAS  Google Scholar 

  • Johnsen SG (1970) Testicular biopsy score count a method for registration of spermatogenesis in human testes: Normal values and results in 335 hypogonadal males. Hormones 1:2–25. https://doi.org/10.1159/000178170

    Article  CAS  Google Scholar 

  • Kurtoglu A, Yavuz R, Evrendilek GA (2014) Characterisation and fate of grayanatoxins in mad honey produced from Rhododendron ponticum nectar. Food Chem 161:47–52

    Article  CAS  Google Scholar 

  • Li W, Ning JZ, Cheng F et al (2018) MALAT1 Promotes Cell Apoptosis and Suppresses Cell Proliferation in Testicular Ischemia-Reperfusion Injury by Sponging MiR-214 to Modulate TRPV4 Expression. Cell Physiol Biochem 46:802–814

    Article  Google Scholar 

  • Maejima H, Kinoshita E, Seyama I, Yamaoka K (2003) Distinct sites regulating grayanotoxin binding and unbinding to D4S6 of Na(v)1.4 sodium channel as revealed by improved estimation of toxin sensitivity. J Biol Chem 278:9464–9471. https://doi.org/10.1074/jbc.M212133200

    Article  CAS  Google Scholar 

  • Mohamadpour M, Noorafshan A, Karbalay-Doust S, Talaei-Khozani T, Aliabadi E (2017) Protective effects of curcumin co-treatment in rats with establishing chronic variable stress on testis and reproductive hormones. Int J Reprod Biomed (Yazd) 15(7):447–452

    Article  CAS  Google Scholar 

  • Onat FY, Yegen BC, Lawrence R, Oktay A, Oktay S (1991) Mad honey poisoning in man and rat. Rev Environ Health 1:4–9

    Google Scholar 

  • Oztasan N, Altinkaynak K, Akcay F, Gocer F, Dane S (2005) Effect of mad honey on blood glucose and lipid levels in rats with streptozocin-induced diabetes. Turk J Vet Anim Sci 29:1093–1096

    CAS  Google Scholar 

  • Pansarasa O, D'Antona G, Gualea MR, Marzani B, Pellegrino M, Marzatico F (2002) Oxidative stress: effects of mild endurance training and testosterone treatment on rat gastrocnemius muscle. Eur J Appl Physiol 87(6):550–555

    Article  CAS  Google Scholar 

  • Peltola V, Huhtaniemi I, Metsa-Ketela T, Ahotupa M (1996) Induction of lipid peroxidation during steroidogenesis in the rat testis. Endocrinology 137(1):105–112

    Article  CAS  Google Scholar 

  • Poon WT, Ho CH, Yip KL et al (2008) Grayanotoxin poisoning from Rhododendron simsii in an infant. Hong Kong Med J 14(5):405–407

    CAS  Google Scholar 

  • Popescu R, Kopp B (2013) The genus Rhododendron: an ethnopharmacological and toxicological review. J Ethnopharmacol 147:42–62

    Article  CAS  Google Scholar 

  • Popescu R, Krupitza G, Kopp B (2009) Novel insights into the mechanism of action of grayanotoxin III. Planta Med:75–PE12

  • Pradhan PM, Niraula SR, Ghimire A et al (2013) Tobacco use and associated factors among adolescent students in Dharan, eastern Nepal: a cross-sectional questionnaire survey. BMJ Open 3:1–7

    Article  Google Scholar 

  • Qiang Y, Zhou B, Gao K (2011) Chemical constituents of plants from the genus Rhododendron. Chem Biodivers 8(5):792–815

    Article  CAS  Google Scholar 

  • Royle NJ, Surai PF, Hartley IR (2001) Maternally derived androgens and antioxidants in bird eggs: complementary but opposing effects? Behav Ecol 12:381–385

    Article  Google Scholar 

  • Rutkowska J, Cichoń M, Puerta M, Gil D (2005) Negative effects of elevated testosterone on female fecundity in zebra finches. Horm Behav 47(5):585–591

    Article  CAS  Google Scholar 

  • Sahin H, Turumtay EA, Yildiz O, Kolayli S (2015) Grayanotoxin-III ddtection and antioxidant activity of mad honey. Int J Food Proper 18:2665–2674

    Article  CAS  Google Scholar 

  • Salman TM, Alagbonsi IA, Olayaki LA et al (2013) Honey increases sperm count in male albino rats by enhancing testosterone production. Biokemistri 25(2):39–44

    Google Scholar 

  • Scott PM, Coldwell BB, Wilberg GS (1971) Grayanotoxins. Occurrence and analysis in honey and a comparison of toxicities in mice. Food and cosmetics. Toxicol 9:179–184

    CAS  Google Scholar 

  • Seyama I, Yamaoka K, Yakehiro M, Yoshioka Y, Morihara K (1985) Is the site of action of grayanotoxin the sodium channel gating of squid axon? Jpn J Physiol 35(3):401–410

    Article  CAS  Google Scholar 

  • Silici S, Atayoğlu TA (2015) Mad honey intoxication: a systematic review on the 1199 cases. Food Chem Toxicol 86:282–290

    Article  CAS  Google Scholar 

  • Silici S, Uluozlu OD, Tuzen M, Soylak M (2008) Assessment of trace element levels in Rhododendron honeys of Black Sea region. Turkey J Hazard Mater 156:612–618

    Article  CAS  Google Scholar 

  • Silici S, Sağdıç O, Ekici L (2010) Total phenolic content, antiradical, antioxidant and antimicrobial activities of Rhododendron honeys. Food Chem 121:238–243. https://doi.org/10.1016/j.foodchem.2009.11.078

    Article  CAS  Google Scholar 

  • Silici S, Sarioglu K, Dogan M, Karaman K (2014a) HPLC-DAD analysis to identify the phenolic profile of Rhododendron honeys collected from different regions in Turkey. Int J Food Proper 17:1126–1135. https://doi.org/10.1080/10942912.2012.698441

    Article  CAS  Google Scholar 

  • Silici S, Yonar E, Sahin H, Atayoğlu AT, Özkök D (2014b) Analysis of grayanatoksin in Rhododendron honey and effect on antioxidant parameters in rats. J Ethnopharmacol 156:155–161

    Article  CAS  Google Scholar 

  • Simon D, Preziosi P, al B-CE (1992) The influence of aging on plasma sex hormones in men: the telecom study. Am J Epidemiol 135(7):783–791

    Article  CAS  Google Scholar 

  • Tajar A, Huhtaniemi IT, O'Neill TW, Finn JD, Pye SR, Lee DM, Bartfai G, Boonen S, Casanueva FFF, Forti G, Giwercman A, Han TS, Kula K, Labrie F, Lean MEJ, Pendleton N, Punab M, Vanderschueren D, Wu FCW, the EMAS Group (2012) Characteristics of androgen deficiency in late-onset hypogonadism: results from the European male aging study (EMAS). J Clin Endocrinol Metab 97(5):1508–1516. https://doi.org/10.1210/jc.2011-2513

    Article  CAS  Google Scholar 

  • Tallent WH, Riethof ML, Horning EC (1957) Studies on the occurrence and structure of acetylandromedol (andromedotoxin). J Am Chem Soc 79:4548–4554

    Article  CAS  Google Scholar 

  • Tam NN, Gao Y, Leung YK, Ho SM (2003) Androgenic regulation of oxidative stress in the rat prostate: involvement of NAD(P)H oxidases and antioxidant defense machinery during prostatic involution and regrowth. Am J Pathol 163(6):2513–2522

    Article  CAS  Google Scholar 

  • Tatli O, Karaca Y, Turkmen S et al (2016) The effect of mad honey on testosterone levels of male rats. Bratisi Med J 117(11):677–680

    Article  CAS  Google Scholar 

  • Traish AM, Guay A, Feeley R, Saad F (2009) The dark side of testosterone deficiency: I. Metabolic syndrome and erectile dysfunction J Androl 30(1):10–22. https://doi.org/10.2164/jandrol.108.005215

    Article  CAS  Google Scholar 

  • Türkmen S, Karagöz Ü, Gunduz A, Türedi S, Metehan A, Yildirim M (2013) The dose-dependent effect of grayanotoxin on the cardiovascular system. Turk J Med Sci 43(5):700–705

    Article  CAS  Google Scholar 

  • Wong J, Youde E, Dickinson B, Hale M (2002) Report of the rhododendron feasibility study. https://www.wildresources.co.uk/reports/rhodo_eng.pdf. Accessed 3 Feb 2018

  • Yilmaz O, Eser M, Sahiner A, Altintop L, Yesildag O (2006) Hypotension, bradycardia and syncope caused by honey poisoning. Resuscitation 68(3):405–408

    Article  Google Scholar 

  • Yilmaz I, Kaya E, Yaykaslı KO, Turker Y (2017) The dose-dependent cardiovascular effects of intravenous grayanotoxin-III in rats. J Dr Behcet Uz Children's Hospital 7(2):121–128

    Google Scholar 

  • Yuki T, Yamaoka K, Yakehiro M, Seyama I (2001) State-dependent action of grayanotoxin I on Na+ channels in frog ventricular myoctes. J Physiol 534(3):777–790

    Article  CAS  Google Scholar 

  • Zhu XD, Bonet B, Knopp RH (1997) 17beta-estradiol, progesterone, and testosterone inversely modulate low-density lipoprotein oxidation and cytotoxicity in cultured placental trophoblast and macrophages. Am J Obstet Gynecol 177(1):196–209

    Article  CAS  Google Scholar 

Download references

Funding

This work was supported by Bozok University Project Coordination Application and Research Center (6602c-TF/17-60).

Author information

Authors and Affiliations

  1. Faculty of Medicine Histology-Embryology Department, Bozok University, Yozgat, Turkey

    Züleyha Doğanyiğit & Emin Kaymak

  2. Seyrani Agricultural Faculty Agricultural Biotechnology Department, Erciyes University, Kayseri, Turkey

    Sibel Silici

  3. Faculty of Medicine Urology Department, Erciyes University, Kayseri, Turkey

    Abdullah Demirtaş

  4. Faculty of Medicine Pharmacology Department, Düzce University, Düzce, Turkey

    Ertuğrul Kaya

Authors
  1. Züleyha Doğanyiğit
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sibel Silici
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Abdullah Demirtaş
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ertuğrul Kaya
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Emin Kaymak
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sibel Silici.

Ethics declarations

The protocol of this study was approved by Ethic Committee, Faculty of Veterinary Medicine, University of Erciyes (protocol no: 16/088).

Additional information

Responsible editor: Philippe Garrigues

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Doğanyiğit, Z., Silici, S., Demirtaş, A. et al. Determination of histological, immunohistochemical and biochemical effects of acute and chronic grayanotoxin III administration in different doses in rats. Environ Sci Pollut Res 26, 1323–1335 (2019). https://doi.org/10.1007/s11356-018-3700-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11356-018-3700-9

Keywords

Search

Navigation