In vitro and In vivo Anticonvulsant Effect of Hydroalcoholic Extracts of Clutia abyssinica in Mice Model

Main Article Content

Samson Sahile Salile
Teferra Abula

Abstract

Background: Epilepsy is a chronic disorder of the brain that affects people of all ages worldwide. In the search of safe and effective antiepileptics traditional treatment practices are one area of research to obtain novel molecules. Research is also needed to validate and standardize the traditional claim. Clutia abyssinica leaves were one of the medicinal plants claimed for use against epilepsy and evil eye and other diseases in different parts of Ethiopia. But there was no scientific research evidence for the claimed use of the plant. Therefore this work was designed to evaluate the anticonvulsant effect of hydroalcoholic extract of Clutia abyssinica leaves.

Methods: The dry residue of the plant extract was used for the test. In vitro 0Mg2+ mice model at dose 0.7 mg/kg of extract, diazepam (3μM) and untreated brain slice groups were used to compare the presence of seizure like event (SLE). In vivo pentylenetetrazol (PTZ) model with 85 mg/kg subcutaneously was used to compare the seizure onset time with two extract doses and diazepam 5 mg/kg. The data was presented with mean± standard error. In maximum electric shock (MES) model 54 mA was passed for 0.2 second transauricularly in mice. The mean time of hind limb extension was recorded for doses 400 mg/kg and 800mg/kg of the extract and 10 mg/kg phenytoin. The means were compared for statistical significance using one way ANOVA post hoc LSD whereas proportions were compared using Fishers exact test with P-value < .05. 

Results: In vitro anticonvulsant tests C. abyssinica extract effect was not statistically significant compared to negative control (P>0.05).A positive control using the known anticonvulsant diazepam (3μM), showed significant anticonvulsant activity (P<0.05). The in vivo PTZ test showed no statistically significant effect in plant extract at all dose levels (P>0.05). In the in vivo MES test the extract of Clutia abyssinica both low and higher dose didn't show statistically significant effect (P>0.05) compared with the negative control. But the extract improved survival (p<0.05). The qualitative secondary metabolite test evidenced the presence of alkaloids, cardiac glycosides, flavanoids, phenols, saponins, sterols and terpeoids in Clutia abyssinica extract.

Conclusion: The hydroalcoholic crude extract result of the C. abyssinica as anticonvulsant is weak based on the models used in this study. For most of the local preparation are mixes of different plants it may have synergistic action with other plants. Or it may have action with other models of chronic epilepsy.

Keywords:
Clutia abyssinica, anti-epileptic, In vitro, In vivo, mice

Article Details

How to Cite
Salile, S. S., & Abula, T. (2021). In vitro and In vivo Anticonvulsant Effect of Hydroalcoholic Extracts of Clutia abyssinica in Mice Model. Journal of Complementary and Alternative Medical Research, 13(4), 8-14. https://doi.org/10.9734/jocamr/2021/v13i430230
Section
Original Research Article

References

Devinsky O, Vezzani A, , Terence J. O’Brien NJ, Curtis M de, Perucca, P. Epilepsy. Nat Rev. 2018;3(18024):4222–31.

Salile SS, Abula T. In vitro and In vivo anticonvulsant effect of hydroalcoholic extract of moringa stenopetala in mice models. J Complement Altern Med Res. 2021;12(3):16–23.

Wereta TB. Ethnobotany of medicinal plants in Erob and Gulomahda districts , Eastern Zone of. PhD thesis Unpublished; 2015.

Tamene S, Addisu D, Debela E. Ethno-medicinal study of plants in Boricha district : Use , preparation and application by traditional healers ,Southern Ethiopia. J Med plant Res. 2020;14(7):343–53.

Wubetu M, Sintayehu M, Abdelwuhab Aeta M, Reta H, Derebe D. Ethnobotany of medicinal plants used to treat various mental illnesses in Ethiopia: A systematic review. J Plant Sci Res [Internet]. 2018 [cited 2019 Apr 12];8(1):9–33.
Available: www.pelagiaresearchlibrary.com

Asfaw T, Helisob T. Assessment of the indigenous knowledge and use of traditional medicinal plants in Wolaita Zone, Southern Ethiopia. Int J Med Plants Nat Prod. 2017;3(1).

Raimondo JV., Heinemann U, de Curtis M, Goodkin HP, Dulla CG, Janigro D, et al. Methodological standards for in vitro models of epilepsy and epileptic seizures. A TASK1-WG4 report of the AES/ILAE translational task force of the ILAE. Epilepsia. 2017;58:40–52.

Löscher W. Critical review of current animal models of seizures and epilepsy used in the discovery and development of new antiepileptic drugs. Seizure [Internet]. 2011;20(5):359–68. Available: http://dx.doi.org/10.1016/j.seizure.2011.01.003

Bialer M, White HS. Key factors in the discovery and development of new antiepileptic drugs. Nature Reviews Drug Discovery. 2010;9:68–82.

Debella A. Manual for phytochemical screening of medicinal plants. Addis Ababa, Ethiopia; 2002.

OECD. Organisation For Economic Cooperation and Development. Guidelines for the Testing of Chemicals, OECD 423. Acute oral toxicity : acute toxic class method. Oecd Guidel Test Chem. 2001;1–14.

Forman CJ, Tomes H, Mbobo B, Burman RJ, Jacobs M, Baden T. Openspritzer : an open hardware pressure ejection system for reliably delivering picolitre volumes. 2017;1–11.

Dreier J, Heinemann U. Regional and time dependent variations of low Mg2+ induced epileptiform activity in rat temporal cortex slices. Exp Brain Res. 1991;3:581–96.
DOI:10.1007/BF00227083

Dreier JP, Zhang CL, Heinemann U, Jp D, C-l Z, Phenytoin HU. Phenytoin, phenobarbital, and midazolam fail to stop status epilepticus-like activity induced by low magnesium in rat entorhinal slices, but can prevent its development. Acta Neurol Scand. 1998 Sep;98(3):154–60.
DOI:10.1111/j.1600-0404.1998.tb07286.x

Raimondo JV, Heinemann U, de Curtis M, Goodkin HP, Dulla CG, Janigro D, et al. Methodological standards for in vitro models of epilepsy and epileptic seizures. A TASK1-WG4 report of the AES/ ILAE Translational Task Force of the ILAE. Epilepsia [Internet]; 2017 [cited 2019 Apr 13];58:40–52.
Available:https://onlinelibrary.wiley.com/doi/pdf/10.1111/epi.13901

Anderson WW, Anderson WW, Lewis D V., Swartzwelder HS, Wilson WA. Magnesium-free medium activates seizure-like events in the rat hippocampal slice. Brain Res. 1986;398(1):215–9.
DOI: 10.1016/0006-8993(86)91274-6

Mody I, Lambert JD, Heinemann U. Low extracellular magnesium induces epileptiform activity and spreading depression in rat hippocampal slices. J Neurophysiol. 1987;57(3):869–88.
DOI: 10.1152/jn.1987.57.3.869

Qaddoumi MG, Ananthalakshmi KVV, Phillips OA, Edafiogho IO, Kombian SB. Evaluation of anticonvulsant actions of dibromophenyl enaminones using in vitro and In vivo seizure models. PLoS One. 2014;9(6): e99770.
DOI: 10.1371/journal.pone.0099770

Hegde K, Thakker SP, Joshi AB, Shastry CS, Chandrashekhar KS. Anticonvulsant activity of Carissa carandas Linn. root extract in experimental mice. Trop J Pharm Res. 2009;8(2):117–25.

Insuasty H, Castro É, Escobar JC, Murillo V, Rodríguez J. Assessment of the anticonvulsant activity of pyrazolo [1,5-a][1,3,5]triazines obtained by synthesis.Rev Colomb cien quím farm. 2014;43(1):22–38.
DOI: 10.15446/rcciquifa.v43n1.45462

Wereta TB. Ethnobotany of medicinal plants in Erob and Gulomahda Districts, Eastern Zone of Tigray Region, Ethiopia. PhD thesis Unpubl. 2015;246–75.

Ragunathan M, Abay SM. Ethnomedicinal survey of folk drugs used in Bahirdar Zuria district, Northwestern Ethiopia. Indian J Tradit Knowl. 2009;8(2):281–4.
Available: http://nopr.niscair.res.in/handle/123456789/3948

SC K, RO O, NM M, MM I, MP N, NM N. Analgesic activity of dichloromethanolic root extract of clutia abyssinica in Swiss Albino Mice. Nat Prod Chem Res. 2017;05(02).

Mergia E, Shibeshi W, Terefe G, Teklehaymanot T. Natural products chemistry & research evaluation of In vivo antitrypanosomal activity of aqueous and methanol leaf extracts of clutia abyssinica (Euphorbiaceae) against trypanosoma congolense field isolate. 2014;2(4).

Meharie BG, Amare GG, Belayneh YM. Evaluation of hepatoprotective activity of the crude extract and solvent fractions of clutia abyssinica (Euphorbiaceae) leaf against ccl4-induced hepatotoxicity in mice. J Exp Pharmacol. 2020;12:137–50.