|
 |
| SHORT COMMUNICATION |
|
|
|
| Year : 2015 | Volume
: 47
| Issue : 4 | Page : 444-446 |
| |
Dual inhibition of acetylcholinesterase and butyrylcholinesterase enzymes by allicin
Suresh Kumar
University School of Biotechnology, GGS Indraprastha University, Dwarka, New Delhi, India
| Date of Submission | 09-Jun-2014 |
| Date of Decision | 17-Jan-2015 |
| Date of Acceptance | 16-Jun-2015 |
| Date of Web Publication | 21-Jul-2015 |
Correspondence Address:
Dr. Suresh Kumar
University School of Biotechnology, GGS Indraprastha University, Dwarka, New Delhi
India
 Source of Support: None, Conflict of Interest: None  | Check |
DOI: 10.4103/0253-7613.161274
Objectives: The brain of mammals contains two major form of cholinesterase enzymes, acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). The dual inhibition of these enzymes is considered as a promising strategy for the treatment of neurological disorder such as Alzheimer's disease (AD), senile dementia, ataxia, and myasthenia gravis. The present study was undertaken to explore the anticholinesterase inhibition property of allicin.
Materials and Methods: An assessment of cholinesterase inhibition was carried out by Ellman's assay.
Results: The present study demonstrates allicin, a major ingredient of crushed garlic (Allium sativum L.) inhibited both AChE and BuChE enzymes in a concentration-dependent manner. For allicin, the IC 50 concentration was 0.01 mg/mL (61.62 μM) for AChE and 0.05 ± 0.018 mg/mL (308.12 μM) for BuChE enzymes.
Conclusions: Allicin shows a potential to ameliorate the decline of cognitive function and memory loss associated with AD by inhibiting cholinesterase enzymes and upregulate the levels of acetylcholine (ACh) in the brain. It can be used as a new lead to target AChE and BuChE to upregulate the level of ACh which will be useful in alleviating the symptoms associated with AD.
Keywords: Acetylcholinesterase, allicin, Alzheimer′s diseases, butyrylcholinesterase
How to cite this article:
Kumar S. Dual inhibition of acetylcholinesterase and butyrylcholinesterase enzymes by allicin. Indian J Pharmacol 2015;47:444-6 |
| » Introduction | |  |
Alzheimer's disease (AD) is the most common cause of dementia in the elderly people is characterized by disturbance of multiple cortical functions, including memory loss, judgement, orientation, comprehension, learning capacity, and language deficit. [1],[2] The major symptoms of all type of dementia are presumed to be related to progressive deterioration of widespread and dense cholinergic innervation of the human cerebral cortex, which contributes to the cognitive and behavioral disturbances in AD, and is also associated with the decreased levels of the neurotransmitter, acetylcholine (ACh), choline acetyltransferase, and acetylcholinesterase (AChE). [3] Apart from AChE, ACh is also hydrolytically destroyed in the brain by another enzyme, butyrylcholinesterase (BuChE). [4] In the healthy brain, AChE is the most important enzyme regulating the levels of ACh, while BuChE plays a minor role. In patient with AD, the levels of AChE activity declines and the activity of BuChE increases and the ratio between BuChE and AChE can change from 0.6 in the normal brain to as high as 11 in the cortical areas affected the disease. [5] These observations lead to dual inhibition strategy of these enzymes has been proposed to increase the efficacy of treatment and broaden the indications.
The medicinal use of garlic and several Indian medicinal plants were demonstrated by more than 1,300 research articles during the past 100 years. [6],[7] This background and the fact that garlic is also described as having immunomodulatory, anti-inflammatory, and antioxidant effects. This focused our interest on the question of whether the known constituents of processed garlic specifically allicin ([R, S]-diallyl disulfide-S-oxide) on inhibition of AChE and BuChE, two important enzymes involved in the breakdown of ACh in the human brain. [8] In the present study, we investigated the inhibitory effects of allicin on AChE and BuChE enzymes.
| » Materials and Methods | | |
Chemicals/Reagents
AChE (EC 3.1.1.7) and BuChE (EC 3.1.1.8) from bovine erythrocytes, acetylthiocholine iodide (ATChI), butyrylthiocholine iodide, 5:5-dithiobis-2-nitrobenzoic acid (DTNB), and sodium bicarbonate were purchased from Sigma, UK. Allicin (Triveni Ltd). The extraction procedure was carried out as described in a previously published study. [9]
Cholinesterase Assays
An assessment of cholinesterase inhibition was carried out in vitro in flat-bottom 96-well microtiter plates using a Ellman's colorimetric method. [10],[11] A typical run consisted of 5 μL of bovine AChE/BuChE solution, at final assay concentrations of 0.03 U/mL; 200 μL of 0.1 M phosphate buffer pH 8; 5 μL of DTNB at a final concentration of 0.3 mM prepared in 0.1 M phosphate buffer pH 7 with 0.12 M of sodium bicarbonate; and 5 μL of the test solution. The reactants were mixed and preincubated for 15 min at 30°C. The reaction was initiated by adding 5 μL of ATChI/BuChI at final concentrations of 0.5 mM. As a control, the inhibitor solution was replaced with buffer. The control and test samples were assayed in triplicate. To monitor any nonenzymatic hydrolysis in the reaction mixture, two blanks for each run were prepared in triplicate. One blank consisted of buffer replacing enzyme and a second blank had buffer replacing substrate. Change in absorbance at 405 nm was measured on a molecular device M2, 96-well plate reader for a period of 6 min at 30°C.
| » Results | | |
Estimation of IC 50 Value of Allicin
The result showed that allicin inhibited the AChE in a concentration-dependent manner at different concentrations tested (0.003 mg/mL to 0.1 mg/mL) [Figure 1], a concentration of 0.1 mg/mL showed maximum inhibition which was 95%. The IC 50 value was 0.01 ± 0.009 mg/mL (61.62 μM) derived from equation of [Figure 1]. This result demonstrated that allicin was a potent AChE inhibitor. In contrast to AChE inhibition, allicin also inhibited BuChE enzyme, but the inhibition was not as strong compared with AChE. The concentration of allicin tested in the range of 0.003-0.1mg/mL, showed percentage inhibition ranging from 16% to 56%, respectively. The maximum inhibition was observed at the concentration of 0.1 mg/mL (56%) [Figure 2]. The IC 50 concentration calculated was 0.05 ± 0.018 mg/mL (308.12 μM) from the equation of [Figure 2]. The results showed that allicin was a weak BuChE inhibitor. | Figure 1: Inhibition of bovine acetylcholinesterase by allicin. Values are expressed as ± standard error of the mean (n = 6). The equation for the line is y = 48.66x + 144.22; R2 = 0.9973
Click here to view |
 | Figure 2: Inhibition of bovine butyrylcholinesterase by different concentration of allicin. Values are expressed as ± standard error of the mean (n = 6). The equation of the line is y = 18.949x2 + 86.651x + 114.59; R2 = 0.9872
Click here to view |
Statistical Analysis
Data obtained were subjected to Windows 7 (Microsoft office) version 2007, mean, ± standard error of the mean, and IC 50 was calculated using Windows 7 (Microsoft Excel software).
| » Discussion | | |
Garlic (Allium sativum) has been used worldwide by Egyption, Chinese, and Indians as a spice, food, and folk medicine for centuries. It has been used to treat various ailments such as heart disease, arthritis, pulmonary complaints, cancer, diarrhea, and worm infestation. [12] Several investigators have reported different medicinal uses of garlic, one of these is its anti-oxidative properties, which not only protect against oxidative damage but also lower the risk of injury to vital molecules and to varying degree may help prevent the onset and progression of age-related neurodegenerative diseases. [13],[14] Studies of senile dementia model in mice showed that aged garlic extract prevented atrophic changes in the frontal brain, improved learning abilities and memory retention, and increased longevity in the senescence-accelerated mouse. [15],[16]
Allicin is reported to have most active lipid-soluble volatile organosulfur biological compound present in the freshly crushed garlic. [17] It was demonstrated earlier that Torpedo californiac AChE when incubated with allicin produced rapid inactivation which was time and concentration dependent. [18] Our results which showed concentration-dependent inhibition of bovine AChE by allicin complementing the previous finding. According to our knowledge, we are for the first time reporting the concentration-dependent inhibition of BuChE by allicin. Although several cholinesterase inhibitors such as tacrine, rivastigmine, and donepezil are being used for management of conditions associated with AD, their side effects have become increasingly noticeable. [19],[20] Therefore, the therapy with these newer derivatives obtained from the natural product that have dual function and lesser adverse effects could be beneficial for AD patients.
In summary, it's demonstrated that allicin strongly inhibit AChE but weakly inhibit BuChE in a concentration-dependent manner. These results may provide an interesting basic contribution with regard to the beneficial effects claimed for garlic and may be of therapeutic value in AD. However, further study on the pharmacological and in vivo behavioral test is needed. Thus, this active compound has a potential to ameliorate the decline of cognitive function and memory loss associated with AD.
Financial Support and Sponsorship
Nil.
Conflicts of Interest
There are no conflicts of interest.
| » References | | |
| 1. | Desgranges B, Baron JC, de la Sayette V, Petit-Taboué MC, Benali K, Landeau B, et al. The neural substrates of memory systems impairment in Alzheimer′s disease. A PET study of resting brain glucose utilization. Brain 1998;121:611-31.  |
| 2. | Grosse DA, Gilley DW, Wilson RS. Episodic and semantic memory in early versus late onset Alzheimer′s disease. Brain Lang 1991;41:531-7. |
| 3. | Lane RM, Potkin SG, Enz A. Targeting acetylcholinesterase and butyrylcholinesterase in dementia. Int J Neuropsychopharmacol 2006;9:101-24. |
| 4. | Giacobini E. Cholinesterase inhibitors: New roles and therapeutic alternatives. Pharmacol Res 2004;50:433-40. |
| 5. | Greig NH, Lahiri DK, Sambamurti K. Butyrylcholinesterase: An important new target in Alzheimer′s disease therapy. Int Psychogeriatr 2002;14:77-91. |
| 6. | Agarwal KC. Therapeutic actions of garlic constituents. Med Res Rev 1996;16:111-24. |
| 7. | Kumar S, Brijeshlata, Dixit S. Screening of Indian spices for inhibitory activity of acetylcholinesterase and butyrlcholinesterase enzymes. Int J Pharma Bio Sci 2012;3:59-65. |
| 8. | Reuter HD, Koch HP, Lawson LD. Therapeutic effects and applications of garlic and its preparations. In: Koch HP, Lawson LD, editors. Garlic: The Science and Therapeutic Application of Allium sativum L. and Related Species. Baltimore: Williams and Wilkins; 1996. p. 135-13. |
| 9. | Block E, Naganathan S, Putman D, Zhao SH. Allium chemistry: HPLC analysis of thiosulfinates from onion, garlic, wild garlic (ramsoms), leek, scallion, shallot, elephant (great-headed) garlic, chive, and chinese chive. Uniquely high allyl to methyl ratios in some garlic samples. J Agric Food Chem 1992;40:2418. |
| 10. | Ellman GL, Courtney KD, Andres V Jr, Feather-Stone RM. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 1961;7:88-95. |
| 11. | Kumar S, Seal CJ, Okello EJ. Kinetics of acetylcholinesterase inhibition by an aqueous extract of Withania somnifera roots. Int J Pharma Sci Res 2011;2:1188-92. |
| 12. | Rivlin RS. Historical perspective on the use of garlic. J Nutr 2001;131:951S-4. |
| 13. | Borek C. Antioxidant and cancer. Sci Med 1997;4:51-62. |
| 14. | Gutteridge JM. Free radicals in disease processes: A compilation of cause and consequence. Free Radic Res Commun 1993;19:141-58. |
| 15. | Nishiyama N, Moriguchi T, Saito H. Beneficial effects of aged garlic extract on learning and memory impairment in the senescence-accelerated mouse. Exp Gerontol 1997;32:149-60. |
| 16. | Moriguchi T, Saito H, Nishiyama N. Anti-ageing effect of aged garlic extract in the inbred brain atrophy mouse model. Clin Exp Pharmacol Physiol 1997;24:235-42. |
| 17. | Ali M, Thomson M, Afzal M. Garlic and onions: Their effect on eicosanoid metabolism and its clinical relevance. Prostaglandins Leukot Essent Fatty Acids 2000;62:55-73. |
| 18. | Millard CB, Shnyrov VL, Newstead S, Shin I, Roth E, Silman I, et al. Stabilization of a metastable state of Torpedo californica acetylcholinesterase by chemical chaperones. Protein Sci 2003;12:2337-47. |
| 19. | Inglis F. The tolerability and safety of cholinesterase inhibitors in the treatment of dementia. Int J Clin Pract Suppl 2002;127:45-63. |
| 20. | Jann MW, Shirley KL, Small GW. Clinical pharmacokinetics and pharmacodynamics of cholinesterase inhibitors. Clin Pharmacokinet 2002;41:719-39. |
[Figure 1], [Figure 2]
| This article has been cited by | | 1 |
Allicin: A review of its important pharmacological activities |
|
| Vivek D. Savairam, Neha A. Patil, Shrikant R. Borate, Mahesh M. Ghaisas, Rajkumar V. Shete | | Pharmacological Research - Modern Chinese Medicine. 2023; : 100283 | | [Pubmed] | [DOI] | | | 2 |
A Review on Garlic as a Supplement for Alzheimer’s Disease: A Mechanistic
Insight into its Direct and Indirect Effects |
|
| Mohammad Mahdi Ghazimoradi, Mozhgan Ghobadi Pour, Ehsan Ghoushi, Hadise Karimi Ahmadabadi, Mahmoud Rafieian-Kopaei | | Current Pharmaceutical Design. 2023; 29(7): 519 | | [Pubmed] | [DOI] | | | 3 |
Prospective Epigenetic Actions of Organo-Sulfur Compounds against Cancer: Perspectives and Molecular Mechanisms |
|
| Shoaib Shoaib, Mohammad Azam Ansari, Mohammed Ghazwani, Umme Hani, Yahya F. Jamous, Zahraa Alali, Shadma Wahab, Wasim Ahmad, Sydney A. Weir, Mohammad N. Alomary, Nabiha Yusuf, Najmul Islam | | Cancers. 2023; 15(3): 697 | | [Pubmed] | [DOI] | | | 4 |
Therapeutic Potential of Allicin and Aged Garlic Extract in Alzheimer’s Disease |
|
| Paola Tedeschi, Manuela Nigro, Alessia Travagli, Martina Catani, Alberto Cavazzini, Stefania Merighi, Stefania Gessi | | International Journal of Molecular Sciences. 2022; 23(13): 6950 | | [Pubmed] | [DOI] | | | 5 |
Phytochemical Compounds Loaded to Nanocarriers as Potential Therapeutic
Substances for Alzheimer’s Disease-Could They be Effective? |
|
| Derya Çiçek Polat, Ayse Esra Karadag, Rabia Edibe Parlar Köprülü, Ioannis D. Karantas, Gökçe Mutlu, Emre Sefik Çaglar, Mehmet Evren Okur, Neslihan Üstündag Okur, Panoraia I. Siafaka | | Current Pharmaceutical Design. 2022; 28(30): 2437 | | [Pubmed] | [DOI] | | | 6 |
Nutrition, Physical Activity, and Other Lifestyle Factors in the Prevention of Cognitive Decline and Dementia |
|
| Ligia J. Dominguez, Nicola Veronese, Laura Vernuccio, Giuseppina Catanese, Flora Inzerillo, Giuseppe Salemi, Mario Barbagallo | | Nutrients. 2021; 13(11): 4080 | | [Pubmed] | [DOI] | | | 7 |
Chemical Composition, Antioxidant, Alpha-glucosidase Inhibitory, Anticholinesterase and Photoprotective Activities of the Aerial Parts of Schinus molle L. |
|
| Mustapha M. Bouhenna, Chawki Bensouici, Latifa Khattabi, Farid Chebrouk, Nabil Mameri | | Current Bioactive Compounds. 2021; 17(6) | | [Pubmed] | [DOI] | | | 8 |
Effects of Black Garlic Extract and Nanoemulsion on the Deoxy Corticosterone Acetate-Salt Induced Hypertension and Its Associated Mild Cognitive Impairment in Rats |
|
| Chun-Yu Chen, Tsung-Yu Tsai, Bing-Huei Chen | | Antioxidants. 2021; 10(10): 1611 | | [Pubmed] | [DOI] | | | 9 |
Allicin, an Antioxidant and Neuroprotective Agent, Ameliorates Cognitive Impairment |
|
| Muhammad Shahid Nadeem, Imran Kazmi, Inam Ullah, Khushi Muhammad, Firoz Anwar | | Antioxidants. 2021; 11(1): 87 | | [Pubmed] | [DOI] | | | 10 |
Pharmacophore-based virtual screening and molecular docking to identify promising dual inhibitors of human acetylcholinesterase and butyrylcholinesterase |
|
| Ana Mércia Silva Mascarenhas, Raquel Bianca Marchesine de Almeida, Moysés Fagundes de Araujo Neto, Géssica Oliveira Mendes, Jorddy Neves da Cruz, Cleydson Breno Rodrigues dos Santos, Mariana Borges Botura, Franco Henrique Andrade Leite | | Journal of Biomolecular Structure and Dynamics. 2021; 39(16): 6021 | | [Pubmed] | [DOI] | | | 11 |
Cholinesterase inhibitory activity, antioxidant properties, and phytochemical composition of
Chlorococcum
sp. extracts
|
|
| Tosin A. Olasehinde, Ademola O. Olaniran, Anthony I. Okoh | | Journal of Food Biochemistry. 2021; 45(3) | | [Pubmed] | [DOI] | | | 12 |
ABCpred: a webserver for the discovery of acetyl- and butyryl-cholinesterase inhibitors |
|
| Aijaz Ahmad Malik, Suvash Chandra Ojha, Nalini Schaduangrat, Chanin Nantasenamat | | Molecular Diversity. 2021; | | [Pubmed] | [DOI] | | | 13 |
Leveraging hallmark Alzheimer’s molecular targets using phytoconstituents: Current perspective and emerging trends |
|
| Prajakta A. Dhage, Archana A. Sharbidre, Sarada P. Dakua, Shidin Balakrishnan | | Biomedicine & Pharmacotherapy. 2021; 139: 111634 | | [Pubmed] | [DOI] | | | 14 |
In silico modeling for dual inhibition of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) enzymes in Alzheimer’s disease |
|
| Vinay Kumar, Achintya Saha, Kunal Roy | | Computational Biology and Chemistry. 2020; 88: 107355 | | [Pubmed] | [DOI] | | | 15 |
Hairy Garlic (Allium subhirsutum) from Sicily (Italy): LC-DAD-MSn Analysis of Secondary Metabolites and In Vitro Biological Properties |
|
| Stefania Sut, Filippo Maggi, Sara Bruno, Natale Badalamenti, Luana Quassinti, Massimo Bramucci, Daniela Beghelli, Giulio Lupidi, Stefano Dall’Acqua | | Molecules. 2020; 25(12): 2837 | | [Pubmed] | [DOI] | | | 16 |
Chemical Constituents and Pharmacological Activities of Garlic (Allium sativum L.): A Review |
|
| Gaber El-Saber Batiha, Amany Magdy Beshbishy, Lamiaa G. Wasef, Yaser H. A. Elewa, Ahmed A. Al-Sagan, Mohamed E. Abd El-Hack, Ayman E. Taha, Yasmina M. Abd-Elhakim, Hari Prasad Devkota | | Nutrients. 2020; 12(3): 872 | | [Pubmed] | [DOI] | | | 17 |
Allicin pharmacology: Common molecular mechanisms against neuroinflammation and cardiovascular diseases |
|
| Feres José Mocayar Marón, Alejandra Beatriz Camargo, Walter Manucha | | Life Sciences. 2020; 249: 117513 | | [Pubmed] | [DOI] | | | 18 |
An In Vitro and In Vivo Cholinesterase Inhibitory Activity of Pistacia khinjuk and Allium sativum Essential Oils |
|
| Peyman Ghajarbeygi, Ashraf Hajhoseini, Motahare-Sadat Hosseini, Anoosheh Sharifan | | Journal of Pharmacopuncture. 2019; 22(4): 231 | | [Pubmed] | [DOI] | | | 19 |
Lichens of Parmelioid Clade as Promising Multitarget Neuroprotective Agents |
|
| Víctor Sieteiglesias, Elena González-Burgos, Paloma Bermejo-Bescós, Pradeep K. Divakar, María Pilar Gómez-Serranillos | | Chemical Research in Toxicology. 2019; 32(6): 1165 | | [Pubmed] | [DOI] | | | 20 |
Valorization of onion solid waste and their flavonols for assessment of cytotoxicity, enzyme inhibitory and antioxidant activities |
|
| Arti Nile, Shivraj Hariram Nile, Doo Hwan Kim, Young Soo Keum, Park Gyun Seok, Kavita Sharma | | Food and Chemical Toxicology. 2018; 119: 281 | | [Pubmed] | [DOI] | | | 21 |
Inhibitory effect of thiacremonone on MPTP-induced dopaminergic neurodegeneration through inhibition of p38 activation |
|
| Chul Ju Hwang, Hee Pom Lee, Dong-Young Choi, Heon Sang Jeong, Tae Hoon Kim, Tae Hyung Lee, Young Min Kim, Dae Bong Moon, Sung Sik Park, Sun Young Kim, Ki-Wan Oh, Dae Yeon Hwang, Sang-Bae Han, Hwa-Jeong Lee, Jin Tae Hong | | Oncotarget. 2016; 7(30): 46943 | | [Pubmed] | [DOI] | | | 22 |
Multitarget strategies in Alzheimer's disease: benefits and challenges on the road to therapeutics |
|
| Michela Rosini, Elena Simoni, Roberta Caporaso, Anna Minarini | | Future Medicinal Chemistry. 2016; 8(6): 697 | | [Pubmed] | [DOI] | |
|
|
|
|
|
|
|
|
