Author(s): Machlusil Husna, Kusworini Handono, Hidayat Sujuti, Aulanni’am Aulanni’am, Afiyfah Kiysa Waafi, Rumman Karimah, Alya Satira


DOI: 10.52711/0974-360X.2024.00192   

Address: Machlusil Husna1,2*, Kusworini Handono3, Hidayat Sujuti4, Aulanni’am Aulanni’am5, Afiyfah Kiysa Waafi6, Rumman Karimah6, Alya Satira6
1Doctoral Program in Medical Science, Faculty of Medicine Universitas Brawijaya, East Java, Indonesia
2Department of Neurology, Faculty of Medicine Universitas Brawijaya, Saiful Anwar General Hospital, Malang, East Java, Indonesia
3Department of Clinical Pathology, Faculty of Medicine Universitas Brawijaya, Saiful Anwar General Hospital, Malang, East Java, Indonesia.
4Department of Ophthalmology, Faculty of Medicine Universitas Brawijaya, Saiful Anwar General Hospital, Malang, East Java, Indonesia.
5Department of Biochemistry, Laboratory of Biochemistry, Faculty of Sciences Universitas Brawijaya, East Malang, Java, Indonesia.
6Master Program in Biomedical Sciences, Faculty of Medicine Universitas Brawijaya, Malang, East Java, Indonesia. *Corresponding Author

Published In:   Volume - 17,      Issue - 3,     Year - 2024

Despite the development of anti-epilepsy drugs, drug-refractory epilepsy still becomes a challenging problem along with an increased incidence of epilepsy. To face that challenge and increase patients’ quality of life, treatment of epilepsy must effectively prevent epileptogenesis, not only symptomatic treatment. AKT signaling pathway was proven to have important roles in epilepsy through its function in the synaptic plasticity, neurogenesis, axon guidance, modulation of the glutamate transporter, and activation of the Ca2+ channel. AKT also activated mTOR signaling pathway as activator of mTORC1 and also effector of mTORC2. Several studies showed the ability of long-term rapamycin treatment to inhibit mTORC2. This study used organotypic hippocampal slice cultures (OHSC) and long-term rapamycin treatment was administered for 3, 5, 8, and 10 days at a dose of 20 nM after induction of epilepsy by low-Mg2+ medium administration for 40 minutes. Low-Mg2+ medium administration induced seizure activity in OHSC showed by significant increase in intracellular Ca2+expressionand also significantly increase AKT activity. After administration of long-term rapamycin treatment AKT activity and intracellular Ca2+expression were significantly reduced. The longer the treatment of rapamycin, the lower the AKT activity and intracellular Ca2+expression. Long-term rapamycin treatment has the potential to become a novel epilepsy drug through its ability to attenuate AKT activity and suppress the seizures proven by lower intracellular Ca2+expression.

Cite this article:
Machlusil Husna, Kusworini Handono, Hidayat Sujuti, Aulanni’am Aulanni’am, Afiyfah Kiysa Waafi, Rumman Karimah, Alya Satira. Long-term Rapamycin Treatment Inhibit AKT Activity and Lower Intracellular Calcium Expression in Organotypic Hippocampal Slice Cultures Model of Epilepsy. Research Journal of Pharmacy and Technology. 2024; 17(3):1232-9. doi: 10.52711/0974-360X.2024.00192

Machlusil Husna, Kusworini Handono, Hidayat Sujuti, Aulanni’am Aulanni’am, Afiyfah Kiysa Waafi, Rumman Karimah, Alya Satira. Long-term Rapamycin Treatment Inhibit AKT Activity and Lower Intracellular Calcium Expression in Organotypic Hippocampal Slice Cultures Model of Epilepsy. Research Journal of Pharmacy and Technology. 2024; 17(3):1232-9. doi: 10.52711/0974-360X.2024.00192   Available on:

1.    Shubhika Jain, Bharti Chogtu, Vybhava Krishna, IshaKhadke. Evaluation of Antiepileptic activity of Mosapride in Albino wistar rats. Research Journal of Pharmacy and Technology. 2021; 14(12): 6364-8. doi: 10.52711/0974-360X.2021.01100
2.    T. Tamilselvan, Arokia Rani C., Ashna Raj, Leena Priya M., Nissy Varghese, Sojan P. Paul. Prescription Analysis of Antiepileptic Drugs in a Tertiary Care Hospital. Asian J. Pharm. Tech. 2018; 8 (1): 43-46 .doi: 10.5958/2231-5713.2018.00007.7
3.    Yogesh R. Joshi, Prabodh V. Sapkale, Pramod P. Patil. Effect of Nimodipine alone and in combination with Gabapentin against Pentylenetetrazole induced Seizures in Mice. Asian J. Pharm. Res. 2018; 8(4): 215-220. doi: 10.5958/2231-5691.2018.00036.9
4.    Beghi E, Giussani G, Nichols E, Abd-Allah F, Abdela J, Abdelalim A, et al. Global, regional, and national burden of epilepsy, 1990–2016: a systematic analysis for the Global Burden of Disease Study 2016. The Lancet Neurology. 201;18(4):357-375.
5.    Campos G, Fortuna A, Falcão A, Alves G. In vitro and in vivo experimental models employed in the discovery and development of antiepileptic drugs for pharmacoresistant epilepsy. Epilepsy Research. 2018; 146: 63-86.
6.    Vinod G. Ugale, Sudhir G. Wadodkar, Chandrabhan T. Chopde. Design, Development and Screening of Some Novel Quinazolinones for Anticonvulsant Activity. Asian J. Research Chem. 2011; 4(11): 1717-1721.
7.    Blair RDG. Temporal lobe epilepsy semiology. Epilepsy Research Treatment. 2012;1–10. doi:10.1155/2012/751510
8.    Rania S. Salah, Hanaa H. Ahmed, Somia H. Abd-Allah, Rasha E. Hassan, Wagdy K.B. Khalil, Ahmed A. Abd-Rabou, Gilane M. Sabry. The Anti-epileptic Efficiency of Mesenchymal Stem Cells Against Pilocarpine Model of Acute Epilepsy. Research J. Pharm. and Tech 2021; 14(3): 1255-1266. doi: 10.5958/0974-360X.2021.00223.7
9.    VinodkumarMugada, Raj Kiran Kolakota. Etiology and Management of Pediatric Seizures: A Descriptive Cross-Sectional Study. Asian J. Res. Pharm. Sci. 2018; 8(4): 236-240. doi: 10.5958/2231-5659.2018.00039.5
10.    N. Sangeetha, U.S. Mahadeva Rao.Hepatotoxic Effect of Sodium Valproate Therapy in Epileptic Children. Research J. Pharma. Dosage Forms and Tech. 2011; 3(4): 135-138.
11.    Griffith JL, Wong M. The mTOR pathway in treatment of epilepsy: a clinical update. Future Neurology. 2018; 13(2): 49-58.
12.    Pun RY, Rolle IJ, LaSarge CL, Hosford BE, Rosen JM, Uhl JD, Schmeltzer SN, et al. Excessive activation of mTOR in postnatally generated granule cells is sufficient to cause epilepsy. Neuron. 2012; 75(6): 1022-1034.
13.    Sha LZ, Xing XL, Zhang D, Yao Y, Dou WC, Jin LR, et al. Mapping the spatio-temporal pattern of the mammalian target of rapamycin (mTOR) activation in temporal lobe epilepsy. PLoS One. 2012; 7(6): e39152.
14.    Nguyen LH, Bordey A. Convergent and divergent mechanisms of epileptogenesis in mTORopathies. Frontiers in Neuroanatomy. 2021; 15: 664695.
15.    Bracho‐Valdés I, Moreno‐Alvarez P, Valencia‐Martínez I, Robles‐Molina E, Chávez‐Vargas L, Vázquez‐Prado J. mTORC1‐and mTORC2‐interacting proteins keep their multifunctional partners focused. IUBMB Life. 2011; 63(10): 896-914.
16.    Valmiki RR, Venkatesalu S, Chacko AG, Prabhu K, Thomas MM, Mathew V, et al. Phosphoproteomic analysis reveals Akt isoform-specific regulation of cytoskeleton proteins in human temporal lobe epilepsy with hippocampal sclerosis. Neurochemistry International. 2020; 134: 104654.
17.    Yang J, Feng G, Chen M, Wang S, Tang F, Zhou J, et al. Glucosamine promotes seizure activity via activation of the PI3K/Akt pathway in epileptic rats. Epilepsy Research. 2021; 175: 106679.
18.    Jiang G, Wang W, Cao Q, Gu J, Mi X, Wang K, et al. Insulin growth factor-1 (IGF-1) enhances hippocampal excitatory and seizure activity through IGF-1 receptor-mediated mechanisms in the epileptic brain.Clinical Science. 2015; 129(12): 1047-1060.
19.    Sharma MK, Jalewa J, Hölscher C. Neuroprotective and anti‐apoptotic effects of liraglutide on SH‐SY 5Y cells exposed to methylglyoxal stress. Journal of Neurochemistry. 2014; 128(3): 459-471.
20.    Viard P, Butcher AJ, Halet G, Davies A, Nürnberg B, Heblich F, et al. PI3K promotes voltage-dependent calcium channel trafficking to the plasma membrane. Nature Neuroscience. 2004; 7(9): 939-946.
21.    Durmus N, Kaya T, Gültürk S, Demir T, Parlak M, Altun A. The effects of L type calcium channels on the electroencephalogram recordings in WAG/RIJ rat model of absence epilepsy. European Review for Medical and Pharmacological Sciences. 2013; 17(9): 1149-54
22.    Mohamed A. Mohamed, Waill A. Elkhateeb, Mohamed A. Taha, Ghoson M. Daba. New Strategies in Optimization of Rapamycin Production by Streptomyces hygroscopicus ATCC 29253. Research Journal of Pharmacy and Technology. 2019; 12(9): 4197-4204. doi: 10.5958/0974-360X.2019.00722.4
23.    Li J, Kim SG, Blenis J. Rapamycin: one drug, many effects. Cell Metabolism. 2014; 19(3): 373-379.
24.    Sarbassov DD, Ali SM, Sengupta S, Sheen JH, Hsu PP, Bagley AF, et al. Prolonged rapamycin treatment inhibits mTORC2 assembly and Akt/PKB. Molecular cell. 2006; 22(2): 159-168.
25.    Blazejczyk M, Macias M, Korostynski M, Firkowska M, Piechota M, Skalecka A. Kainic acid induces mTORC1-dependent expression of Elmo1 in hippocampal neurons. Molecular Neurobiology. 2017;54(4):2562-2578.
26.    Koyama R, Muramatsu R, Sasaki T, Kimura R, Ueyama C, Tamura M, et al. A low-cost method for brain slice cultures. Journal of Pharmacological Sciences. 2007; 104(2): 191-194.
27.    Buckmaster PS, Lew FH. Rapamycin suppresses mossy fiber sprouting but not seizure frequency in a mouse model of temporal lobe epilepsy.Journal of Neuroscience. 2011; 31(6): 2337-2347.
28.    Liu J, Saponjian Y, Mahoney MM, Staley KJ, Berdichevsky Y. Epileptogenesis in organotypic hippocampal cultures has limited dependence on culture medium composition. PloS One. 2017; 12(2): e0172677.
29.    Mody I, Lambert JDC, Heinemann U. Low extracellular magnesium induces epileptiform activity and spreading depression in rat hippocampal slices. Journal of Neurophysiology. 1987; 57(3): 869-888.
30.    Mele M, Vieira R, Correia B, De Luca P, Duarte FV, Pinheiro PS, et al. Transient incubation of cultured hippocampal neurons in the absence of magnesium induces rhythmic and synchronized epileptiform-like activity. Scientific Report. 2021; 11(1): 1-14.
31.    Pisani A, Bonsi P, Martella G, De Persis C, Costa C, Pisani F, Bernardi G, Calabresi P. Intracellular calcium increase in epileptiform activity: modulation by levetiracetam and lamotrigine. Epilepsia. 2004; 45(7): 719-728.
32.    Vadim Tsyvunin, SergiyShtrygol, Mariia Mishchenko, Iryna Ryzhenko, Diana Shtrygol, Denis Oklei. Low-Dose Digoxin is Associated with Anticonvulsant Effect Enhancement of Classical Antiepileptic Drugs in the Electro-Induced Seizures in Mice. Research Journal of Pharmacy and Technology. 2022; 15(9): 4241-7. doi: 10.52711/0974-360X.2022.00713
33.    Kirkland AE, Sarlo GL, Holton KF. The role of magnesium in neurological disorders. Nutrients. 2018; 10(6): 730.
34.    DeLorenzo RJ, Pal S, Sombati S. Prolonged activation of the N-methyl-d-aspartate receptor–Ca2+ transduction pathway causes spontaneous recurrent epileptiform discharges in hippocampal neurons in culture. Proceeding of the National Academy of Sciences. 1998; 95(24): 14482-14487.
35.    Parag Jain, Anand Surana, Ravindra Pandey, Shiv Shankar Shukla. Epilepsy: A Neurological Cramp. Research J. Pharmacology and Pharmacodynamics. 2013; 5(1):  1-5.
36.    Zheng F, Zhou X, Moon C, Wang H. Regulation of brain-derived neurotrophic factor expression in neurons. International Journal of Physiology, Pathophysiology and Pharmacology. 2012; 4(4): 188. PMID: 23320132
37.    Sliwa A, Plucinska G, Bednarczyk J, Lukasiuk K. Post-treatment with rapamycin does not prevent epileptogenesis in the amygdala stimulation model of temporal lobe epilepsy. Neuroscience Letter. 2012; 509(2): 105-109.
38.    van Vliet EA, Forte G, Holtman L, den Burger JC, Sinjewel A, de Vries HE, et al. Inhibition of mammalian target of rapamycin reduces epileptogenesis and blood–brain barrier leakage but not microglia activation. Epilepsia. 2012; 53(7): 1254-1263.
39.    Nguyen LH, Brewster AL, Clark ME, Regnier‐Golanov A, Sunnen CN, Patil VV, et al. mTOR inhibition suppresses established epilepsy in a mouse model of cortical dysplasia. Epilepsia. 2015; 56(4): 636-646.
40.    Ljungberg MC, Sunnen CN, Lugo JN, Anderson AE, D’Arcangelo G. Rapamycin suppresses seizures and neuronal hypertrophy in a mouse model of cortical dysplasia. Disease Model and Mechanism. 2009; 2(7-8): 389-398.
41.    Buckmaster PS, Ingram EA, Wen X. Inhibition of the mammalian target of rapamycin signaling pathway suppresses dentate granule cell axon sprouting in a rodent model of temporal lobe epilepsy. Journal of Neuroscience. 2009; 29: 8259–8269.

Recomonded Articles:

Research Journal of Pharmacy and Technology (RJPT) is an international, peer-reviewed, multidisciplinary journal.... Read more >>>

RNI: CHHENG00387/33/1/2008-TC                     
DOI: 10.5958/0974-360X 

56th percentile
Powered by  Scopus

SCImago Journal & Country Rank

Recent Articles


Not Available