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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Date
2021-06
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Addis Ababa University
Abstract
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.
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Keywords
Pterolobium stellatum, Moringa stenopetala, Clutia abyssinica, in vitro, in vivo, anticonvulsant, mice