Combined Treatment of Levetiracetam and Mesenchymal Stem Cells Reverses the Biochemical Aberrations in the Acute Phase of Epilepsy Induced by Pilocarpine in Rats

Abstract

In spite of the enormous evolution of the novel anti-seizure medications, about one-third of epilepsy patients stay resistant to the existing therapeutic drugs. Stem cells have provoked hopeful for treating diverse neurologic diseases comprises epilepsy. The rational of this investigation was to appraise the therapeutic intervention of a combination of levetiracetam (LEV) with rodent adipose-derived mesenchymal stem cells (ADMSCs) or rodent bone marrow-derived mesenchymal stem cells (BMMSCs) in counteracting pilocarpine-induced acute epilepsy in rats. In this research, the isolation and preparation of ADMSCs and BMMSCs from male albino rats were carried out. The identification of ADMSCs and BMMSCs was performed morphologically in the culture by using the inverted microscope and by the detection of the cell surface profile by using the flow cytometry technique. The induction of acute epilepsy was achieved by intraperitoneal injection of a single dose of pilocarpine (380 mg/kg b.wt). This study was conducted on fifty six adult male albino rats which were assigned into seven equal groups (8 rats/group); Group (1): Control, Group (2): Epileptic, Group (3): Epileptic + LEV (300 mg/kg b.wt daily for 12 weeks by gastric intubation with an oral gavage), Group (4): Epileptic + ADMSCs (single dose of ADMSCs; 3 × 106 cells/rat; intravenously), Group (5): Epileptic + BMMSCs (single dose of BMMSCs 3 × 106 cells/rat; intravenously), Group (6) : Epileptic + ADMSCs + LEV and Group (7) Epileptic + BMMSCs + LEV. After the end of the experimental period (12 weeks), all rats were tested by the mean electric shock (MES) test to prove the presence of strong seizures in the epileptic group and the modulation of these seizures after treatments. Then, the rats were decapitated and the whole brain of each rat was dissected into two halves, the first half was used for the quantitative determination of GABA, glutamate, dopamine, bFGF, BDNF, IL-6 and TNF-α, while the second half was fixed in formalin saline for histological investigation. The findings of the present work demonstrated that the morphological appearance of the isolated MSCs manifests spindle-shape. The flow cytometric analysis showed that the isolated MSCs are positive for CD90 and negative for CD14 and CD45. The homing of MSCs in the brain tissue of the treated rats was verified by their staining with the fluorescent dye. The recordings of the MES indicated the presence of strong seizures in the epileptic rats which were ameliorated after treatment with LEV, ADMSCs, BMMSCs. The level of brain GABA decreased significantly in the epileptic rats, whereas the level of brain glutamate, dopamine, bFGF, BDNF, IL-6 and TNF-α increased significantly; these alterations were improved after treatment with LEV or ADMSCs or BMMSCs. The histological examination of the brain tissue of the epileptic rats showed great histopathological alterations which were amended by the different treatment options. The combined treatment of either ADMSCs or BMMSCs with LEV displayed superior advantageous effect versus the single use of each type of cell in combating the acute phase of epilepsy. In conclusion, the outcomes of the present approach disclosed that the combined treatment of either ADMSCs or BMMSCs with the antiepileptic drug LEV has synergistic effect in alleviation of the behavioral and biochemical aberrations as well as brain histological deformation during the acute phase of epilepsy.