Anticonvulsant Effect of Pterlobium stellatum (Leaves), Moringa stenopetala (Root) And Clutia abyssinica (Leaves) Traditionally Used for Treatment of Epilepsy in Ethiopia Using Mice Model

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Addis Ababa University


Background :Epilepsy is the third leading contributor to the global burden of disease for neurological disorders and affects 65 million people worldwide. Although the current antiepileptic drugs achieve symptomatic seizure relief they do not prevent or reverse the pathological process that underlies human epilepsy. Drug resistant epilepsy is also one of the most important unmet needs in the daily management of epilepsy. These currently unmet needs provide a roadmap for the development of more effective antiseizure drugs, as well as for disease modifying and antiepileptogenic drugs especially from plants. This study was conducted to investigate the anticonvulsant activity of the crude hydroalcoholic extracts of 3selected plants Pterlobium stellatum ( leaves), Moringa stenopetala (root) and Clutia abyssinica (leaves). These all are used for treatment of epilepsy by traditional healers in different parts of Ethiopia. Additionally the solvent fractions of the Pterlobium stellatum were tested for anticonvulsant activity as the crude extract showed positive response in all the models used. Methods :.The dry residues of the plant extracts were used for test in different doses. Male balb c mice were used for in vivo study and for in vitro study P14-P21 of C57BL16 mice were used. In vitro mice model of hippocampal slice with 0Mg 2+ was used and the extracts were tested at the 0.7mg/kg concentration. Diazepam 3µM was positive control and DMSO as negative control. In in vivo PTZ and MES mice models 400mg/kg and 800mg/kg of each test extract were used for efficacy test in as positive control Diazepam 5mg/kg and phenytoin 10mg/kg were used respectively. The negative control was 2% tween 80 .Fisher's exact test was used to analyze proportions and ANOVA with post hoc LSD to test means. The tests were conducted after the ethical clearance was obtained from the ethical committee of Addis Ababa University and university of Cape Town. Qualitative test and quantitative tests were used to determine the secondary metabolites in the plants. And ultraperformance liquid chromatography-mass spectrometer (UPLCMS) tests were used to characterize plant constituents in Pterlobium stellatum crude extract and its fractions. Results :In the in vitro study the hydroalcoholic extract of P.stellatumand M. stenopetalaat 0.7mg/ml had a statistically significant anticonvulsant activity compared to negative control(P<0.05). The hydroalcoholic extract of C.abyssinicaat 0.7mg/ml didn't show statistically significant effect compared to negative control (P>0.05). A positive control, diazepam(3µM), showed statistically significant anticonvulsant activity compared with negative control (P<0.05).When we compare the in vitro activity of different solvent fractions of P. stellatum,the chloroform and water fractions at 0.7mg/ml were also shown to have significant anticonvulsant activity as compared to negative control (P<0.05). The petether and butanol fractions activities were not statistically significant compared to negative control (P>0.05). Pterolobium stellatum hydroalcoholic extract shown that dose dependent and statistically significant anticonvulsant activity with PTZ model(P<0.05).Whereas the activity of M. stenopetala and C. abyssinica hydroalcoholic extracts were not statistically significant (P>0.05) in in vivo PTZ model. The in vivo PTZ test has also revealed the chloroform fraction and the water fraction of Pterolobium stellatum to have anticonvulsant effect(P<0.05) compared with the negative control. Whereas the pet ether and butanol fractions shown activity which was not statistically significant with negative control(P>0.05).The effect of the diazepam was statistically significant with the negative control and all test extracts (P<0.05). Pterolobium stellatum and M. stenopetala hydroalcoholic extracts shown statistically significant anticonvulsant activity with MES model in both lower and higher doses(P<0.05). The C. abyssinica hydroalcoholic extracts activity at the given doses were not statistically significant (P>0.05).The in vivo MES test has also revealed the chloroform fraction to have anticonvulsant effect at both doses (P<0.05). The water fraction at 400mg/kg dose shown anticonvulsant effect compared with the negative control(P<0.05). The pet ether and butanol fractions shown activity which was not statistically significant at the given doses (P>0.05).The effect of the phenytoin was statistically significant with the negative control as well as compared with other tested extract doses(P<0.05). The qualitative and quantitative analysis indicated the presence of different plant secondary metabolites in hydromethanolic extracts of the three plants. In Pterolobium stellatum the UPLCMS analysis indicated also the presence of gallic acid, ellagic acid, kaempferol, myricitrin, isoquercitrin and quercitirin in the crude extract. Of these gallic acid and ellagic acid were found in chloroform fraction. In the water fraction ellagic acid, kaempferol, myricitrin and isoquercitrin were found. Conclusion: The results demonstrated that Pterolobium stellatum has anticonvulsant effect. The hydroalcoholic and chloroform and water fraction of Pterolobium stellatum demonstrated effect in both in vitro and in vivo MES and PTZ models of epilepsy. The crude extract of Moringa stenopetala has also shown to have anticonvulsant effect both in the in vitro and in vivo MES models. But was negative on PTZ model. The traditional use of both herbs for treatment of epilepsy can be supported by the finding of this study. However ,C. abyssinica didn't show anticonvulsant activity at the tested doses in the models used in this study and its traditional use for treatment of epilepsy is not supported according to the findings of this study.



Pterolobium stellatum, Moringa stenopetala, Clutia abyssinica, in vitro, in vivo, anticonvulsant, mice