Anticonvulsant Effect of a Formulated Nigerian-Made Modified Atkins Diet in Mice
Description
Modified Atkins diet has been in use (particularly in developed nations) for a very long time in the management of intractable epilepsy. This research proposed that, a formulated modified Atkins diet using cheap food materials available in Nigeria will also have beneficial effect in seizure management in laboratory animals. The diet was successfully formulated. However, it failed to protect against seizure in all the acute seizure models employed. The diet however, significantly increased seizure threshold/plasticity in the chronic kindling test. The formulated MAD also enhanced learning memory, and caused an increase in motor coordination. Correlation analysis between seizure scores and levels of some relevant biomarkers revealed that the anti-epileptogenic activity of the diet is not likely to be due to its effect on inflammatory cytokines. All the data were analyzed using the SPSS software version 23.0. Different statistical tools- Mixed design ANOVA, one way ANOVA, independent t-test, kruskall-Wallis test etc as shown in the legend for each table of raw data - were employed for the data analysis.
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Chemicals and Equipment Pentylenetetrazole (Sigma Aldrich, UK), Diazepam (Juhel Pharmaceuticals, Nigeria), Phenobarbitone (Sterop laboratories, Belgium), Phenytoin (Pfizer Global Pharmaceuticals, USA), Strychnine, atropine sulphate monohydrate, pilocarpine and Sodium Valproate (Sanofi-Aventis, UK). Equipment include Ugo basile electroshock machine (Model 7800 with corneal electrodes) connected directly to a power source, stopwatch and observation cages, Morris Water Maze apparatus, Camera, Stopwatch, EDTA bottles, Disposable gloves, Face masks and ELISA Kits (to measure levels of TNF-α, BDNF and IL-1β and IL-6) 1. Maximum Electroshock Test (MEST): Method of Swinyard and Kufferberg (1985) was adopted. Predetermined electroshock parameters (Current, 10mA; Pulse frequency, 50Hz; Shock duration, 6seconds and Pulse width, 16ms) were used to induce seizure via ear clip electrode in each mouse. 2. Pentylenetetrazol-Induced Seizure: The protocol described by Swinyard et al., (1989) was used. 3. Strychnine-Induced Seizure in Mice: The method described by Porter et al., (1984) was used. 4. Pilocarpine-Induced Status Epilepticus (SE): SE was induced in accordance with the method described by Lenz et al., 2017. After pilocarpine injection, onset of seizure and the behavioral seizure score were documented using the modified Racine’s stages (Lüttjohann et al., 2009) (0 = no seizures, 1 = freezing, 2 = single twitches, 3 = orofacial seizures, 4 = clonic seizures, 5 = tonic seizures, and 6 = death. 5. PTZ-Induced Kindling: Protocol described by Shimada and Yamagata (2018) was adopted. 6. Positional Sense Test: This test was conducted 24hours after the kindling test. Method described by Holmes (2008) was adopted. 7. Morris Water Maze: Protocol described by Qian et al. (2004) was adopted with slight modifications. 8. Biochemical Analysis: The levels of the various biomarkers were assayed for using ELISA kits and adopting manufacturers instruction. The brainless mice were subsequently incinerated.