 |
 |
 |
 |
 |
 |
Sains Malaysiana 52(3)(2023): 837-850
http://doi.org/10.17576/jsm-2023-5203-12
Neuroprotective
Effects of Ocimum basilicum L. var. thyrsiflora on Scopolamine-Induced Non-Spatial Memory Deficits in Rats
(Kesan Neuropelindung Ocimum basilicum L. var. thyrsiflora pada Kemerosotan Ingatan Bukan Reruang Aruhan Skopolamina Tikus)
NOR
SYAFINAZ YAAKOB1, MOHAMED ALI SEYED2, MANALI HANITI
MOHD-ZAHID3, HENG-HUI TEE1, SHU-YUN YAP1,
SEK-WEN HO1, YUSOF KAMISAH4 & NORAZRINA AZMI1,*
1Centre for Drug and Herbal Development,
Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif, Bandar
Tun Razak, 56000 Kuala Lumpur, Federal Territory, Malaysia
2Department of Biochemistry, Faculty
of Science, University of Tabuk, Tabuk 71491, Kingdom of Saudi Arabia
3Department of Chemical Pathology,
School of Medical Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian,
Kelantan Darul Naim, Malaysia
4Department of Pharmacology, Faculty
of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif, Bandar Tun
Razak, 56000 Kuala Lumpur, Federal Territory, Malaysia
Diserahkan:
13 Oktober 2022/Diterima: 10 Februari 2023
Abstract
Pharmacological studies indicated that Ocimum
basilicum L. var. thyrsiflora has numerous therapeutic potentials.
The aim of this study was to investigate the neuroprotective action of O.
basilicum leaf extract against scopolamine-induced non-spatial memory
deficits in rats and to determine the changes in mRNA expressions of genes
implicated in cognition and neuroprotection. O. basilicum leaves were
extracted with 80% ethanol and verified for the presence of rosmarinic acid
using high performance liquid chromatography method. Male Wistar rats were
treated orally with either O. basilicum or the positive control piracetam for
14 days prior to the injection of 0.5 mg/kg scopolamine on the day of the novel
object discrimination (NOD) test. Hippocampi were collected at the end of the
test. mRNA expression of nicotinic
acetylcholine α7 subunit (NA7), muscarinic M1 receptor (M1), neuronal nitric oxide synthase
(nNOS), and 5-hydroxytryptamine receptor 3A (HTR3A) genes in the hippocampi
were analyzed using qPCR method. The presence of rosmarinic acid in the plant
extract was detected at chromatogram peak of Rt=16.891. NOD test results
indicated that the lower dose of O. basilicum (200 mg/kg) significantly
(p<0.05) reversed scopolamine-induced memory deficits in rats similar
to the effects of piracetam. In addition, O. basilicum at the same dose
alleviated the increase in mRNA expressions of the NA7, M1, nNOS, and HTR3A
genes induced by scopolamine. The present findings suggest that O.
basilicum is potentially neuroprotective in preventing
memory impairment through alleviation of scopolamine-induced changes in
hippocampal mRNA expression implicated in cognition and neuroprotection.
Keywords: Cognition; gene expression; memory;
neuroprotection; Ocimum basilicum
Abstrak
Kajian farmakologi
menunjukkan bahawa Ocimum basilicum L. var. thyrsiflora mempunyai banyak potensi
terapeutik. Matlamat kajian ini adalah untuk mengkaji kesan neuropelindung
ekstrak daun O. basilicum terhadap kemerosotan ingatan bukan reruang
aruhan skopolamina pada tikus dan untuk menentukan perubahan dalam ekspresi
mRNA gen yang terlibat dalam kognisi dan perlindungan saraf. Daun O.
basilicum telah diekstrak dengan 80% etanol dan kehadiran asid rosmarinik
disahkan menggunakan kaedah kromatografi cecair berprestasi tinggi. Tikus
Wistar jantan diberi O. basilicum atau piracetam sebagai kawalan positif
secara oral selama 14 hari sebelum suntikan skopolamina 0.5 mg/kg diberikan
pada hari ujian diskriminasi objek novel (NOD). Hipokampus dikumpul pada akhir
ujian tersebut. Ekspresi mRNA subunit α7 nikotinik asetilkolin (NA7), reseptor
M1 muskarinik (M1), nitrik oksida sintase neuron (nNOS), dan gen reseptor
5-hidroksitriptamina 3A (HTR3A) dalam hippokampus dianalisis menggunakan kaedah
qPCR. Kehadiran asid rosmarinik dalam ekstrak tumbuhan dikesan pada puncak
kromatogram Rt=16.891. Keputusan ujian NOD menunjukkan bahawa dos O.
basilicum yang lebih rendah (200 mg/kg) memulihkan kemerosotan ingatan yang
disebabkan oleh skopolamina dengan ketara (p<0.05) pada tikus serupa dengan
kesan piracetam. Di samping itu, O. basilicum pada dos yang sama
mengurangkan peningkatan dalam ekspresi mRNA gen NA7, M1, nNOS dan HTR3A yang
diaruh oleh skopolamina. Hasil kajian ini menunjukkan bahawa O. basilicum mempunyai potensi neuropelindung dalam mencegah kemerosotan ingatan melalui
pengurangan kesan yang disebabkan oleh skopolamina dalam ekspresi mRNA
hipokampus yang terlibat dalam kognisi dan perlindungan saraf.
Kata kunci: Ekspresi gen; ingatan; kognisi; Ocimum
basilicum; perlindungan saraf
RUJUKAN
Al-Snafi, A.E. 2021a. Chemical constituents
and pharmacological effects of Ocimum basilicum - A review. International
Journal of Pharmaceutical Research 13(2): 2997-3013.
Al-Snafi,
A.E. 2021b. Medicinal plants possessed beneficial therapeutic effects in
Alzheimer’s disease and memory deficits. GSC Biological and Pharmaceutical
Sciences 17(02): 008-033.
Azmi, N., Norman, C., Spicer, C.H.
& Bennett, G.W. 2006. Effects of a neurotensin analogue (PD149163) and
antagonist (SR142948A) on the scopolamine-induced deficits in a novel object
discrimination task. Behavioural Pharmacology 17(4): 357-362.
Badiana, S.L.N., Ngah, E.,
Touo’yem, W.S.N., Sidiki, N., Bum, E.N. 2021. Ocimum basilicum L.
(Lamiaceae) leaves aqueous extract improve learning and memory in the
monosodium glutamate-induced neurotoxicity model of Alzheimer's disease through
attenuating brain oxidative damage in experimental mice. GSC Biological and
Pharmaceutical Sciences 17: 204-212.
Barnes,
J.M., Costall, B., Coughlan, J., Domeney, A.M., Gerrard, P.A., Kelly, M.E.,
Naylor, R.J., Onaivi, E.S., Tomkins, D.M. & Tyers, M.B. 1990. The effects
of ondansetron, a 5-HT3 receptor antagonist, on cognition in rodents and
primates. Pharmacology Biochemistry
and Behavior 35(4): 955-962.
Bartolini, L., Casamenti, F. & Pepeu, G.
1995. Aniracetam restores object recognition impaired by age, scopolamine, and
nucleus basalis lesions. Pharmacology Biochemistry Behavior 53: 277-283.
Berić, T., Nikolić, B.,
Stanojević, J., Vuković-Gačić, B. &
Knežević-Vukčević, J. 2008. Protective effect of basil (Ocimum basilicum L.) against oxidative
DNA damage and mutagenesis. Food and Chemical Toxicology 46(2): 724-732.
Bertaina-Anglade, V.B., Enjaunes, E., Morillon, D.
& Rochele, C.D. 2006. The object recognition test in rats and mice: A
simple and rapid model in safety pharmacology to detect amnesic properties of a
new chemical entity. Journal of Pharmacology and Toxicology Methods 54(2): 99-105.
Besheer, J., Short, K.R. & Bevins,
R.A. 2001. Dopaminergic and cholinergic antagonism in a novel-object detection
task with rats. Behavior Brain Research 126: 211-217.
Bora, K.S., Arora, S. & Shri, R. 2011. Role of Ocimum
basilicum L. in prevention of ischemia and reperfusion-induced cerebral
damage, and motor dysfunctions in mice brain. Journal of Ethnopharmacology 137(3): 1360-1365.
Bradley, S.J., Bourgognon, J.M., Sanger, H.E.,
Verity, N., Mogg, A.J., White, D.J., Butcher, A.J., Moreno, J.A., Molloy, C.,
Macedo-Hatch, T., Edwards, J.M., Wess, J., Pawlak, R., Read, D.J., Sexton,
P.M., Broad, L.M., Steinert, J.R., Mallucci, G.R., Christopoulos, A., Felder,
C.C. & Tobin, A.B. 2017. M1 muscarinic allosteric modulators slow prion
neurodegeneration and restore memory loss. Journal of Clinical Investigation 127(2): 487-499.
Buhot, M.C., Martin, S. & Segu, L. 2000. Role
of serotonin in memory impairment. Annals of Medicine 32(3):
210-221.
Carli,
M., Luschi, R. & Samanin, R. 1997. Dose-related impairment of spatial
learning by intrahippocampal scopolamine: Antagonism by ondansetron, a 5-HT3 receptor antagonist. Behavioural
Brain Research 82(2): 185-194.
Cheng, Q.
& Yakel, J.L. 2015. Activation of α7 nicotinic acetylcholine receptors
increases intracellular cAMP levels via activation of AC1 in hippocampal
neurons. Neuropharmacology 95: 405-414.
Cheng, Q.
& Yakel, J.L. 2014. Presynaptic α7 nicotinic acetylcholine receptors
enhance hippocampal mossy fiber glutamatergic transmission via PKA activation. Journal
of Neuroscience 34(1): 124-133.
Chidamabaram,
S.B., Pandian, A., Sekar, S., Haridass, S., Vijayan, R., Thiyagarajan, L.K.,
Ravindran, J., Rao, H., Raghavendran, B. & Kamarul, T. 2015. Sesame indicum, a nutritional
supplement, elicits antiamnesic effect via cholinergic pathway in scopolamine
intoxicated mice. Environmental Toxicology 31(12): 1955-1963.
Deutsch, J.A. 1971. The cholinergic
synapse and the site of memory. Science 174: 788-794.
Dodart, J.C., Mathis, C. & Ungerer, A. 1997.
Scopolamine-induced deficits in a two-trial object recognition task in mice. Neuroreports 8(5):
1173-1178.
Ennanceur, A. &
Meliani, K. 1992a. Effects of physostigmine and scopolamine on rats’
performances in object-recognition and radial maze tests. Psychopharmacology (Berl). 109: 321-330.
Ennaceur, A. & Meliani, K. 1992b. A new
one-trial test for neurobiological studies of memory in rats. III. Spatial vs
non-spatial working memory. Behavior Brain Research 51: 83-92.
Ennanceur, A. & Delacour, J. 1988. A new
one-trial test for neurobiological studies of memory in rats. Behavioral
data. Behavior Brain Research 31(1): 47-59.
Fakhfouri, G., Rahimian,
R., Dyhrfjeld-Johnsen, J., Zirak, M.R. & Beaulieu, J.M. 2019. 5-HT3 receptor
antagonists in neurologic and neuropsychiatric disorders: The iceberg still
lies beneath the surface. Pharmacology
Reviews 71(3):
383-412.
Falsafi,
S.K., Deli, A., Hoger, H., Pollak, A. & Lubec, G. 2012. Scopolamine
administration modulates muscarinic, nicotinic and NMDA receptor systems. PLoS
ONE 7(2): e32082.
Giridharan, V.V., Thandavarayan, R.A., Mani,
V., Dundapa, T.A., Watanabe, K. & Konishi, T. 2011. Ocimum sanctum Linn. leaf extracts inhibit acetylcholinesterase and
improve cognition in rats with experimentally induced dementia. Journal of
Medicinal Food 14(9): 912-919.
Grayer,
R.J., Kite, G.C., Goldstone, F.J., Bryan, S.E., Paton, A. & Putievsky, E.
1996. Infraspecific taxonomy and essential oil chemotypes in sweet basil, Ocimum
basilicum. Phytochemistry 143: 1033-1039.
Haam, J.
& Yakel, J.L. 2017. Cholinergic modulation of the hippocampal region and
memory function. Journal of Neurochemistry 142: 111-121.
Harrell, A.V. & Allan, A.M.
2003. Improvements in hippocampal-dependent learning and decremental attention
in 5-HT3 receptor overexpressing mice. Learning & Memory 10(5): 410-419.
Hasanein, P. & Mahtaj, A.K. 2015.
Ameliorative effect of rosmarinic acid on scopolamine-induced memory impairment
in rats. Neuroscience Letters 585: 23-27.
Jayasinghe, C.J., Gotoh, N., Aoki, T. &
Wada, S. 2003. Phenolics composition and antioxidant activity of sweet basil (Ocimum
basilicum L.). Journal of Agri and
Food Chemistry 51(15): 4442-4449.
Joshi, H.
& Parle, M. 2006. Evaluation of nootropic potential of Ocimum sanctum Linn. in mice. Indian Journal of Experimental
Biology 44(2): 133-136.
Konar,
A., Gupta, R., Shukla, R.K., Maloney, B., Khanna, V.K., Wadhwa, R., Lahiri,
D.K. & Thakur M.K. 2019. M1 muscarinic receptor is a key target of
neuroprotection, neuroregeneration and memory recovery by i-Extract from Withania
somnifera. Scientific Reports 9(1): 13990.
Koutroumanidou,
E., Kimbaris, A., Kortsaris, A., Bezirtzoglou, E., Polissiou, M.,
Charalabopoulos, K. & Pagonopoulou, O. 2013. Increased seizure latency and
decreased severity of pentylenetetrazol-induced seizures in mice after
essential oil administration. Epilepsy Research and Treatment 2013:
532657.
Kumagai,
Y., Kikushima, M., Nakai, Y., Shimojo, N. & Kunimoto, M. 2004. Neuronal
nitric oxide synthase (NNOS) catalyzes one-electron reduction of
2,4,6-trinitrotoluene, resulting in decreased nitric oxide production and
increased nNOS gene expression: Implication for oxidative stress. Free
Radical Biology and Medicine 37(3): 350-357.
Kumar, N.
& Kela, A.K. 2004. Ondansetron amelioration of scopolamine induced
cognitive deficits in three-panel runway apparatus in rats. Indian Journal
Experimental Biology 42(9): 919-921.
Kwon,
S.H., Lee, H.K., Kim, J.A., Hong, S.I., Kim, H.C., Jo, T.H., Young, I.P., Chong, K.L., Yong, B.K., Seok, Y.L. & Choon, G.J. 2010. Neuroprotective effects of chlorogenic acid on scopolamine-induced
amnesia via anti-acetylcholinesterase and anti-oxidative activities in mice. European
Journal of Pharmacology 649(1-3): 210-217.
Levey, A.I. 1996. Muscarinic acetylcholine receptor
expression in memory circuits: Implications for treatment of
Alzheimer disease. Proceedings of the National Academy
of Sciences 93(24): 13541-13546.
Lochner, M. & Thompson, A.J. 2016. The
muscarinic antagonists scopolamine and atropine are competitive antagonists at
5-HT3 receptors. Neuropharmacology 108:
220-228.
Lu, C., Wang, Y., Xu, T., Li, Q., Wang, D., Zhang,
L., Fan, B., Wang, F. & Liu, X. 2018. Genistein ameliorates scopolamine-induced
amnesia in mice through the regulation of the cholinergic neurotransmission,
antioxidant system and the ERK/CREB/BDNF signaling. Frontier Pharmacology 9:
1153.
Marisco, P.C., Carvalho, F.B., Rosa, M.M., Girardi, B.A.,
Gutierres, J.M., Jaques, J.A., Salla, A.P., Pimentel, V.C., Schetinger, M.R.,
Leal, D.B., Mello, C.F. & Rubin, M.A. 2013. Piracetam prevents
scopolamine-induced memory impairment and decrease of NTPDase, 5’-nucleotidase
and adenosine deaminase activities. Neurochemical Research 38(8):
1704-1714.
Mohamed,
R.A., Abdallah, D.M., El-Brairy, A.I., Ahmed, K.A. & El-Abhar, H.S. 2021.
Palonosetron/Methyllycaconitine deactivate hippocampal microglia 1,
inflammasome assembly and pyroptosis to enhance cognition in a novel model of
neuroinflammation. Molecules 26(16): 5068.
Mohd-Zahid,
M.H., Jalil, J., Chan, K.M. & Azmi, N. 2018. Neuroprotective effects of Ocimum
basilicum extract against hydrogen peroxide-induced oxidative stress in
SK-N-SH neuroblastoma cells. Sains Malaysiana 47(9): 2129-2139.
Moura,
A.C., Lazzari, V.M., Agnes, G., Almeida, S., Giovenardi, M. & Veiga, A.B.
2014. Transcriptional expression study in the central nervous system of rats:
What gene should be used as internal control? Einstein (São Paulo) 12(3): 336-341.
Müller, W.E., Eckert, G.P. & Eckert, A. 1999.
Piracetam: Novelty in a unique mode of action. Pharmacopsychology 32(1):
2-9.
Othman,
M.S., Khaled, A.M., Al-Bagawi, A.H., Fareid, M.A., Ghany, R.A., Habotta, O.A.
& Abdel Moneim, A.E. 2021. Hepatorenal protective efficacy of flavonoids
from Ocimum basilicum extract in
diabetic albino rats: A focus on hypoglycemic, antioxidant, anti-inflammatory
and anti-apoptotic activities. Biomedicine and Pharmacotherapy 144:
112287.
Paul, V. & Ekambaram, P. 2011. Involvement of
nitric oxide in learning and memory processes. Indian Journal of Medical
Research 133(5): 471-478.
Pfaffl,
M.W., Horgan, G.W. & Dempfle, L. 2002. Relative expression software tool
(REST) for group-wise comparison and statistical analysis of relative
expression results in real-time PCR. Nucleic Acids Research 30(9): e36.
Piller,
N., Decosterd, I. & Suter, M.R. 2013. Reverse transcription quantitative
real-time polymerase chain reaction reference genes in the spared nerve injury
model of neuropathic pain: Validation and literature search. BMC Research
Notes 6: 266.
Pitsikas, N., Rigamonti, A.E., Cella, S.G.
& Muller, E.E. 2003. The 5-HT1A receptor antagonist WAY 100635 improves
rats’ performance in different models of amnesia evaluated by the object
recognition task. Brain Research 983: 215-222.
Rahman,
W., Bauer, C.S., Bannister, K., Vonsy, J.L., Dolphin, A.C. & Dickenson,
A.H. 2009. Descending serotonergic facilitation and the antinociceptive effects
of pregabalin in a rat model of osteoarthritic pain. Molecular Pain 5:
45.
Ramakers,
C., Ruijter, J.M., Deprez, R.H. & Moorman, A.F. 2003. Assumption-free
analysis of quantitative real-time polymerase chain reaction (PCR) data. Neuroscience
Letters 339(1): 62-66.
Rameshrad,
M., Salehian, R., Fathiazad, F., Hamedeyazdan, S., Garjani, M. &
Maleki-Dizaji, N. 2015. The effects of Ocimum
basilicum ethanol extract on carrageenan induced paw inflammation in rats. Pharmaceutical
Sciences 20(4): 149-156.
Riedel,
W.J., Klaassen, T., Griez, E., Honig, A., Menheere, P. & van Praag, H.M.
2002. Dissociable hormonal, cognitive and mood responses to neuroendocrine
challenge: Evidence for receptor-specific serotonergic dysregulation in
depressed mood. Neuropsychopharmacology 26(3): 358-367.
Sarahroodi,
S., Esmaeili, S., Mikaili, P., Hemmati, Z. & Saberi, Y. 2012. The effects
of green Ocimum basilicum hydroalcoholic extract on retention and
retrieval of memory in mice. Ancient Science of Life 31(4): 185-189.
Seifhosseini, S., Mehrdad, J., Ali, M. &
Aazami, N.S. 2011. The effect of scopolamine on avoidance memory and
hippocampal neurons in male wistar rats. Basic and Clinical
Neuroscience 3: 9-15.
Seyed, M.A., Ayesha, S.,
Azmi, N., Al-Rabae, F.M.,
Al-Alawy, A.I., Al-Zahrani, O.R. & Hawsawi, Y. 2021. The neuroprotective
attribution of Ocimum basilicum: A review on the prevention and
management of neurodegenerative disorders. Future Journal of
Pharmaceutical Sciences 7: 139.
Sheffler, Z.M.,
Reddy, V. & Pillarisetty, L.S. 2021. Physiology, neurotransmitters. StatPearls.
Treasure Island (FL): https://www.ncbi.nlm.nih.gov/books/NBK539894/
Singh,
V., Krishan, P. & Shri, R. 2018. Improvement of memory and neurological deficit
with Ocimum basilicum L. extract after ischemia reperfusion induced
cerebral injury in mice. Metabolic Brain Disease 33(4): 1111-1120.
Singh,
V., Kahol, A., Singh, I.P., Saraf, I. & Shri, R. 2016. Evaluation of
antiamnesic effect of extracts of selected Ocimum species using in
vitro and in vivo models. Journal of Ethnopharmacology 193:
490-499.
Srivastava, S., Cahill, D.M., Conlan, X.A.
& Adholeya, A. 2014. A novel in vitro whole plant system for
analysis of polyphenolics and their antioxidant potential in cultivars of Ocimum
basilicum. Journal of Agri and Food
Chemistry 62(41):
10064-10075.
Sudweeks,
S.N., Hooft, J.A. & Yakel, J.L. 2002. Serotonin 5-HT(3) receptors in rat
CA1 hippocampal interneurons: Functional and molecular characterization. The
Journal of Physiology 544(Pt 3): 715-726.
Taglialatela, G., Hogan, D., Zhang, W.R.,
Dineley, K.T. 2009. Intermediate- and long-term recognition memory deficits
inTg2576 mice are reversed with acute calcineurin inhibition. Behavior Brain
Research 200: 95-99.
Vlase,
J.L., Benedec, D., Hanganu, D., Damian, G., Csillag, I., Sevastre, B., Mot,
A.C., Silaghi-Dumitrescu, R. & Tilea, I. 2014. Evaluation of antioxidant
and antimicrobial activities and phenolic profile for Hyssopus officinalis, Ocimum basilicum and Teucrium chamaedrys. Molecules 19(5): 5490-5507.
Wallace,
T.L., Ballard, T.M., Pouzet, B., Riedel, W.J. & Wettstein, J.G. 2011. Drug
targets for cognitive enhancement in neuropsychiatric disorders. Pharmacology
Biochemistry and Behavior 99(2): 130-145.
Warburton,
E.C., Koder, T., Cho, K., Massey, P.V., Duguid, G., Barker, G.R.I., Aggleton,
J.P., Bashir, Z.I. & Brown, M.W. 2003. Cholinergic neurotransmission is
essential for perirhinal cortical plasticity and recognition memory. Neuron 38: 987-996.
Winblad, B. 2005. Piracetam: A review of
pharmacological properties and clinical uses. CNS Drug Reviews 11(2):
169-182.
Xiao, J.,
Li, S., Sui, Y., Wu, Q., Li, X., Xie, B., Zhang, M. & Sun, Z. 2014. Lactobacillus
casei-01 facilitates the ameliorative effects of proanthocyanidins
extracted from lotus seedpod on learning and memory impairment in
scopolamine-induced amnesia mice. PLoS ONE 9(11): e112773.
Yamamoto,
N., Yoshida, A., Taki, Y., Onoue, S., Kagawa, Y. & Yamada, S. 2011.
Up-regulation of nicotinic and muscarinic receptor mRNA in rat bladder by
repeated administration of nicotine in relation to the pharmacokinetics. Life
Sciences 89(9-10): 343-348.
Yan, Z.,
Flores-Hernandez, J. & Surmeier, D.J. 2001. Coordinated expression of
muscarinic receptor messenger RNAs in striatal medium spiny neurons. Neuroscience 103(4): 1017-1024.
Yanpallewara, S.U., Raib, S., Kumarb, M. &
Acharya, S.B. 2004. Evaluation of antioxidant and neuroprotective effect of Ocimum sanctum on transient cerebral
ischemia and long-term cerebral hypoperfusion. Pharmacology Biochemistry and
Behavior 79(1): 155-164.
*Pengarang untuk surat-menyurat;
email: azrina.azmi@ukm.edu.my
|
|||
|
