Abstract
Nicotiana tabacum, Datura stramonium, and Carica papaya are plants that are on the high trend as substitutes to conventional psychoactive substances due to their legality and the difference in the experiences they offer. The present study was aimed at comparing the neuromodulatory and toxicological potentials of the afore-named plants to an illicit psychoactive plant, Cannabis sativa. Consequently, the effects of the alkaloid extract of the plants were evaluated on critical neuronal enzymes of the monoaminergic, cholinergic, and purinergic (sodium/potassium adenosine triphosphatase (Na+/K+-ATPase), ecto-5’-nucleotidase [eNTDase], and ecto-nucleoside triphosphate diphosphohydrolase [E-NTPDase]) systems of neurotransmission, reactive oxygen species (ROS) production, and lipid peroxidation in rat brain tissue homogenate ex vivo. Plants’ alkaloids were prepared by solvent extraction method. Results revealed that the extracts inhibited the enzymes in a concentration-dependent manner. However, C. sativa had the highest inhibition of monoamine oxidase (MAO), eNTDase, and E-NTPDase activities, while D. stramonium had the highest cholinesterase and Na+/K+-ATPase activity inhibition, and ROS production and lipid peroxidation. In conclusion, D. stramonium altered critical neuronal enzymes significantly more than the illicit plant of abuse, while Nicotiana tabacum showed no significant difference in comparison to C. sativa. Therefore, the use of these plants as drugs should be discouraged.
Similar content being viewed by others
Explore related subjects
Discover the latest articles, news and stories from top researchers in related subjects.Avoid common mistakes on your manuscript.
Introduction
As old as human history, plant-based drugs have their usage in religious rites and medicines by various ancient traditions. These plants are commonly referred to as psychoactive plants due to their ability to affect the mind or alter the state of consciousness, by directly or indirectly modulating neurotransmitter systems, such as the dopaminergic, cholinergic, purinergic, serotonergic, noradrenalinegic, and Ƴ-butyric; pathways; and their receptors (Cunha-Oliveira et al. 2008). Toxicity and dependency on these plants have been associated with alkaloids which have the potentials of inducing hallucinogenic, stimulatory, or euphoric effects (Graziano et al. 2017). However, recorded abuse was dated to the 19th century during the Ching Dynasty of China resulting in the enactment of laws during the 1961 Single Convention on Narcotic Drugs with further reviews during the Conventions on Psychotropic substances in 1971 and Illicit Traffic in Narcotic Drugs and Psychotropic Substances in 1988 (Feng et al. 2017). Recent data showed that abuse of psychoactive plants is not limited to Cannabis sativa (marijuana), Papayer somniferum (opium poppy), Erythroxylon coca (cocaine), magic mushrooms, etc., but that many other plant-based drugs which pose threat to public health are elusive. They were thus defined by UNODC (United Nation Office on Drugs and Crime) as new psychoactive substances (NPS) of plant origin or new psychoactive plants (NPP), of which about 20 were reported among the total 230 psychoactive substances reported in-between January 2005 and December 2012 by European Monitoring Centre for Drugs and Drug Addiction (EMCDDA) (Ujváry 2014). These new psychoactive plants have thus served as alternatives to the illicit plant-based drugs due to the false perception of them being legal and organic, thereby safe. As much as the surge prevalence and abuse of these plants, data on toxicological, pharmacological, and clinical effects are limited. Putting into consideration a large number of plants of abuse, there is, therefore, a need for techniques that capture multiple mechanisms of action of the plants in comparison with the conventional psychoactive substances while screening ex vivo. Commonly abused of these new psychoactive plants in Nigeria are N. tabacum, D. stramonium, and C. papaya (male and female). Most plant extract IC50 values usually occur close to the estimated brain concentrations following recreational abuse of these plants, indicating their efficiency in ex vivo approach for toxicological and classification screening. This study aims to compare the neuromodulatory and toxicological potentials of these plants of abuse to C. sativa L, an illicit psychoactive plant.
Materials and methods
Materials
Drugs and chemicals
ATPase, ADPase, AMPase, adenosine, acetic acid, methanol, acetylthiocholine, and butyrylthiocholine iodide were purchased from Sigma (St Louis, MO, USA) and Merck (Darmstadt, Germany). All other chemicals used in the present study were of analytical grade.
Methods
Sample collection and preparation
Fresh leaves of C. sativa, N. tabacum, and C. papaya (male and female), and seeds of Datura stramonium were obtained in Akure South Local Government of Ondo State, Nigeria. The plants were identified and authenticated at the Centre for Research and Development, Federal University of Technology, Akure, Nigeria. The leaves and seeds were carefully cleansed of stalks, debris, and other unwanted materials, sundry, and blend into powder using an electric blender.
Preparation of alkaloid extracts
Alkaloid extracts of C. sativa, N. tabacum, D. stramonium, and C. papaya (male and female) were prepared using the method of Harborne (1998), as modified by Ademiluyi et al. (2016b). Typically, 100 g of powdered samples were defatted for 24 h with n-hexane. A total of 200 mL of 10% acetic acid in ethanol were thereafter added to the defatted samples and shaken vigorously; the mounting pressure was vented and allowed to stand for 24 h to permit sufficient extraction. The mixtures were filtered with a muslin cloth, filtered with Whatman No.1 filter paper, and concentrated using a rotary evaporator (Laborota 4000 Efficient, Heidolph, Germany) at 45 °C. Concentrated ammonium hydroxide dropwise was added to the concentrated filtrate to achieve a good precipitate. The precipitate was harvested after allowing the whole solution to settle down to obtain the alkaloid extracts, which were stored frozen for further analysis at 4 °C.
Animal handling
Twenty (20) male albino Wistar rats weighing between 210 and 212 g were obtained from the Animal House of the Department of Biochemistry, Federal University of Technology, Akure, Ondo State, Nigeria. Their handling and use were approved by the Animal ethical committee, Centre for Research and Development (CERAD) of the Federal University of Technology, Akure with the ethical number FUTA/ETH/2020/016. The animals were kept in plastic cages and housed at room temperature (25–27 °C), relative humidity (60–70%), and controlled light cycle (12-h dark/12-h light) for 2 weeks with free access to commercial rat chow and water ad libitum before use.
Preparation of tissue homogenate
The rats were anesthetized with isofluorane before rapid decapitation, after which the whole brain tissue was rapidly removed, rinsed with cold saline to remove blood stains. Thereafter, the brain was weighed and then homogenized with Tris-HCl buffer (1/5 w/v) (pH 7.4) with 10-up-and-down strokes at about rev/min in a Teflon glass (Mexxcare, mc14 362, Aayu-shi Design Pvt. Ltd. India) homogenizer (Adefegha et al. 2015). Afterward, the homogenate was centrifuged at 3000 rpm for 10 min at 4 °C using a refrigerated centrifuge (KX3400C, KENXIN Intl. Co., Hong Kong). The supernatant was obtained and used for biochemical assays, while the pallet was discarded.
Monoamine oxidase activity assay
The inhibitory potentials of C. sativa, N. tabacum, D. stramonium, and C. papaya (male and female) alkaloid extracts on MAO activity were assayed according to the method of Turski et al. (1973) with slight modification. To the varying concentrations of the alkaloid extracts (0–25 µg/mL) were added 150 µL of 0.025 M phosphate buffer (pH 7.0), 12.5 mM semicarbazide, 10 mM benzylamine, and 100 µL of brain homogenate, glacial acetic acid was then added to the mixture after 30 min and incubated in boiling water for 3 min and then centrifuged. One (1) ml of the supernatant was mixed with 1.25 ml of benzene and 1 ml of 2, 4-DNPH, and incubated at room temperature for 10 min. One (1) ml of the benzene layer was then mixed with 1 ml of 0.1 N NaOH. The alkaline layer was decanted and incubated for 10 min at 80 °C, the absorbance read at 450 nm. The MAO inhibitory potential of the samples was calculated as stated below;
where:Absref = absorbance of reference
Abssam = absorbance of sample
Cholinesterase activity assay
The inhibitory potentials of alkaloid extract from C. sativa, N. tabacum, D. stramonium, and C. papaya (male and female) on cholinesterase (acetylcholinesterase [AChE] and butyrylcholinesterase [BChE]) activities were assayed according to the method of Perry et al. (2000). To the varying concentrations of the alkaloid extracts (0–15 µg/mL) were added 30 µL of 10 mM 5, 5′-dithio-bis (2-nitrobenzoic) acid (DTNB), 30 µL of brain homogenate in 0.1 M phosphate buffer (pH 8.0), and 0.1 M phosphate buffer (pH 8.0). The reaction mixture was incubated at 25 °C for 20 min, after which acetylthiocholine iodide (for AChE) or butyrylthiocholine iodide (for BChE) was added as substrate, the absorbance read at 450 nm and the AChE/BChE inhibitory potential of the samples was calculated as stated below:
Where:Absref = absorbance of reference
Abssam = Absorbance of Sample
Ecto-nucleoside triphosphate diphosphohydrolase activity assay
The inhibitory potentials of C. sativa, N. tabacum, D. stramonium, and C. papaya (male and female) alkaloid extracts on ecto-nucleoside triphosphate diphosphohydrolase (E-NTPDase) (using ATP and ADP as substrate) activity were assayed (Schetinger et al. 2007). A total of 150 µL of the substrate (ATP or ADP) was added to the varying concentrations of the alkaloid extracts (0–25 µg/mL), and 100 µL of CaCl2 and Tris-HCl buffer (pH 7.4) to make a total volume of 350 µL. The reaction mixture was incubated for 10 min at 37 °C, after which 150 µL of brain homogenate, incubated for a further 20 min, and 500 µL of 10% of trichloroacetic acid was then added. The reaction mixture was allowed to chill on ice for 10 min, and the supernatant was used to assay for released inorganic phosphate (Pi) according to the method of Fiske and Subbarow (1925). The ATPase/ADPase activity was calculated as stated below and expressed as percentage ATP/ADP inhibition:
Where:Absref = absorbance of reference
Abssam = absorbance of sample
Ecto-5′ nucleotidase assay
The inhibitory potentials of C. sativa, N. tabacum, D. stramonium, and C papaya (male and female) alkaloid extracts on ecto-5-nucleotidase (eNTDase) activity were assayed according to the method of Heymann et al. (1984). To the varying concentrations of the alkaloid extracts (0–25 µg/mL) were added 150 µL AMP, 100 µL of MgCl2, and Tris-HCl buffer (pH 7.4) to make a total volume of 350 µL. The reaction mixture was incubated for 10 min at 37 °C, after which 150 µL of brain homogenate, incubated for a further 20 min, and 500 µL of 10% of trichloroacetic acid was then added. The reaction mixture was allowed to chill on ice for 10 min, and the supernatant was used to assay for released inorganic phosphate (Pi) according to the method of Fiske and Subbarow (1925). The inhibition of ecto-5-nucleotidase was calculated as stated below:
Where:Absref = absorbance of reference
Abssam = absorbance of sample
Na+/K+-ATPase activity assay
The inhibitory potentials of C. sativa, N. tabacum, D. stramonium, and C. papaya (male and female) alkaloid extracts on Na+/K+-ATPase activities were assayed according to the method of Wyse et al. (2000). A total of 100 µL Na+/K+-ATPase substrate (Ouabain), buffer (pH 7.4) (containing 120 mM Tris-HCl, 0.4 mM EDTA, 200 mM NaCl, 20 mM KCl, and 24 mM MgCl2), 50 µL of homogenate, and 50 µL of 1 MM ouabain were added to the varying concentrations of the alkaloid extracts (0–15 µg/mL) to make a total volume of 200 µL. The reaction was initiated by adding 50 µL of adenosine triphosphate (ATP), incubated at 37 °C for 30 min, and terminated by adding 50 µL of 10% trichloroacetic acid (TCA). The reaction mixture was allowed to chill on ice for 10 min, and the supernatant was used to assay for released inorganic phosphate (Pi) according to the method of Fiske and Subbarow (1925). The percentage Na+/K+-ATPase inhibition was calculated as stated below:
Where:Absref = absorbance of reference
Abssam = absorbance of sample
Reactive oxygen species assay
The formation of reactive oxygen species potentials of C. sativa, N. tabacum, D. stramonium, and C. papaya (male and female) were estimated as H2O2 equivalent according to the method of Oboh et al. (2018) using the n-n-diethyl-para-phenylenediamine (DEPPD) reagent. -n-Diethyl-para-phenylenediamine (DEPPD) reagent was added to the varying concentrations of the alkaloid extracts (0–15 µg/mL), and the mixture was incubated for 5 min at 37 °C. Absorbance was measured using a spectrophotometer at 505 nm, while the standard calibration curve of H2O2 was used to quantify reactive oxygen species levels and expressed in unit/mg protein, wherein 1 unit equals 1 mg H202/L.
Thiobarbituric acid-reactive substance formation assay
The formation of thiobarbituric acid-reactive substances (TBARS) by C. sativa, N. tabacum, D. stramonium, and C papaya (male and female) were assayed according to the method of Ohkawa et al. (1979) with slight modification. A total of 50 µL of brain homogenate, 300 µL of 8.1% sodium dodecyl sulfate (SDS), 500 µL HCl/acetic acid (pH 3.4), and 500 µL of thiobarbituric acid (TBA) were added to the varying concentrations of the alkaloid extracts (0–25 µg/mL), and the mixture was incubated for 1 h at 100 °C. The formed TBARS was quantified using a spectrophotometer at 532 nm and calculated as TBARS produced.
Data analysis
All data were analyzed and expressed as values representing triplicate readings (mean ± standard deviation), while the level of significant difference (P ≤ 0.05) was by one-way analysis of variance (ANOVA) using Duncan multiple test. IC50 values (effective concentration that caused 50% inhibition) were calculated with non-linear regression analysis, while the statistical analyses were carried out via version 5 of the GraphPad Prism.
Results
Typically, Fig. 1 shows the effects of the alkaloid extracts of C. sativa, N. tabacum, D. stramonium, and C. papaya (male and female) on the activity of MAO. The alkaloid extracts inhibited MAO activity in a concentration dependent manner. As revealed by IC50 value, C. sativa had the highest (12.34 ± 0.27 μg/ml) value closely followed by that of Nicotiana tabacum (12.99 ± 0.31 μg/ml), D. stramonium (17.47 ± 0.0.35 μg/ml), C. papaya (20.76 ± 0.42 μg/ml), while C. papaya female (28.96 ± 0.58 μg/ml) has the least when the effects of alkaloid extracts from C. sativa, N. tabacum, D. stramonium, and C. papaya (male and female) were compared on ex vivo activity of MAO. More so, as shown in Figs. 2 and 3, it was observed that alkaloid extract from C. sativa, N. tabacum, D. stramonium, and C. papaya (male and female) inhibited the cholinesterase (AChE and BChE) activity in a concentration dependent manner. Finding from this study revealed that AChE and BChE activity was significantly (P < 0.05) inhibited by alkaloid extracts from C. sativa, N. tabacum, D. stramonium, and C. papaya (male and female). Nevertheless, as revealed by IC50 value, D. stramonium had highest ((AChE = 5.48±0.19 μg/ml; BChE = 12.10 ± 0.25 μg/ml)) inhibitory effect, while C. sativa ((AChE = 6.60 ± 0.20 μg/ml; BChE = 14.48 ± 0.29 μg/ml)), N. tabacum ((AChE = 11.30 ± 0.29 μg/ml; BChE = 21.89 ± 0.42 μg/ml)), C. papaya male ((AChE = 14.28 ± 0.33 μg/ml; BChE = 34.40 ± 0.67 μg/ml)), while C. papaya female ((AChE = 16.19 ± 0.37 μg/ml; BChE = 40.93 ± 0.80 μg/ml)) had the least inhibitory effect on ex vivo activity of AChE and BChE. Figures 4 and 5, respectively, depict the effect of alkaloid extract from C. sativa, N. tabacum, D. stramonium, and C. papaya (male and female) on ex vivo activity of ecto-nucleoside triphosphate diphosphohydrolase (E-NTPDase and ADPase). The results revealed that alkaloid extracts exhibited dose response inhibitory effect on ex vivo activity ecto-nucleoside triphosphate diphosphohydrolase with C. sativa ((ATPase = 13.51 ± 0.30 μg/ml; ADPase = 18.19 ± 0.36 μg/ml)) had highest inhibitory effect followed by D. stramonium ((ATPase = 14.23 ± 0.32 μg/ml; ADPase = 18.73 ± 0.37 μg/ml)), N. tabacum ((ATPase = 17.12 ± 0.35 μg/ml; ADPase = 24.29 ± 0.48 μg/ml)), C. papaya male ((ATPase = 22.64 ± 0.43 μg/ml; ADPase = 50.49 ± 1.02 μg/ml)), while C. papaya female ((ATPase = 25.27 ± 0.47 μg/ml; and ADPase = 53.79 ± 1.08 μg/ml)) had the least when their effects were compared. The present study also observed the same trend for ecto-5-nucleotidase (eNTDase) activity inhibition by the plant extracts (Fig. 6). C. sativa had the highest modulatory effect (AMPase = 14.91 ± 0.34 μg/ml), followed by D. stramonium (AMPase = 15.11 ± 0.34 μg/ml), then N. tabacum (AMPase = 18.39 ± 0.43 μg/ml) and C. papaya male (AMPase = 20.45 ± 0.44 μg/ml) and finally C. papaya female (AMPase = 22.77 ± 0.50 μg/ml). Also, Na+/K+-ATPase activity inhibition was also observed to be affected by the alkaloid extracts in a concentration-dependent manner (Fig. 7). Considering the effects of the extracts using 50% modulatory inhibitory effect on Na+/K+-ATPase activity, D. stramonium (Na+/K+-ATPase = 8.36 ± 0.19 μg/ml) had higher inhibitory effect than even C. sativa (Na+/K+-ATPase = 9.44 ± 0.20 μg/ml), an illicit psychoactive substance. Meanwhile, N. tabacum Na+/K+-ATPase = 11.21 ± 0.23 μg/ml), Carica papaya male (Na+/K+-ATPase = 19.89 ± 0.38 μg/ml), and C. papaya female (Na+/K+-ATPase = 21.40 ± 0.42 μg/ml) values were also observed. Finally, the present study investigated the effect of the alkaloid extracts on H2O2 production and lipid peroxidation (Figs. 8 and 9). H2O2 production and lipid peroxidation levels were significantly (P < 0.05) elevated in the present study with D. stramonium, having the highest values of 8.18 ± 0.15 μg/ml and 13.77 ± 0.25 μg/ml, respectively. Our control, C. sativa also caused H2O2 production and lipid peroxidation levels 8.94 ± 0.16 μg/ml and 14.97 ± 0.28 μg/ml, respectively, which is not significantly different from that observed of D. stramonium. Furthermore, ROS production and lipid peroxidation levels of N. tabacum (9.71 ± 0.18 μg/ml, and 18.57 ± 0.34 μg/ml), C. papaya male (29.48 ± 0.58 μg/ml, and 39.98 ± 0.73 μg/ml) and C. papaya female (34.55 ± 0.67 μg/ml, and 48.95 ± 0.91 μg/ml) were also observed.
Discussion
A plant-based psychoactive substance is a substance, which has its primary effect directed at the state of consciousness through the direct or indirect modulation of neurotransmitter systems, such as the dopaminergic, cholinergic, and purinergic systems and their receptors (Cunha-Oliveira et al. 2008). The present study observed that C. sativa had the highest MAO inhibition of which can be due to the presence of delta-9-tetrahydrocannabinol (THC), its active psychoactive constituent as observed by (Baggio et al. 2014), though less potent compared to that observed with iproniazid, a standard non-selective monoamine oxidase inhibitor (IC50 = 0.72 μg/mL) by Zhi et al. (2016). Close to the illicit substance is N. tabacum which can be relatively due to the presence of its active constituents—nicotine and beta-carboline alkaloids (harman and nor-harman), which are present in tobacco smoke (Herraiz and Chaparro 2005). Typically, these two plant-based psychoactive substances have always been on the top chart of the most abused substances (World Drug Report 2015) with their high level of addiction and high range of acceptance by users cutting across the global world (Hasin et al. 2013; World Drug Report 2015). More so, a considerable inhibition of MAO was observed in D. stramonium, though its high level of toxicity and lethal effects as observed by Ademiluyi et al. (2016a, b) and Ogunmoyole et al. (2019) can be the underline factor marring its addictiveness and dependence. Meanwhile, the level of amino acids tyrosine, and phenylalanine, and tryptophan, the precursors of norepinephrine and serotonin, respectively, in C. papaya (male and female) might be the underline factor of their MAO inhibition as observed by Parle (2011). Furthermore, the higher IC50 value observed in the male C. papaya compares to that of the female Carica papaya might be the underline factor for its preference over the female counterpart by abusers as observed in folklore.
As much as the neurological importance of the activities of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) are, their high inhibition as observed in the present study can ultimately result in excess accumulation of the choline at the synaptic cleft in living organisms when unchecked, causing overstimulation of the cholinergic and postsynaptic neurons and resultantly leading to neuronal death and/or even the organism’s death (Ademiluyi et al. 2016b; Rodrigues et al. 2011). Notably, the present study observed that D. stramonium had higher AChE and BChE inhibitory values compared to C. sativa, an illicit psychoactive substance. The high level of cholinesterase activity inhibition by D. stramonium correlates with previous studies that associated the high inhibition with the presence of alkaloids such as atropine, hyoscyamine, and scopolamine. Furthermore, N. tabacum and C. papaya (male and female) were also observed to pose a significant inhibitory effect, with C. papaya female exhibiting a lesser potency compared to the male species.
The inhibition of E-NTPDase activities as observed in the present study could lead to impairments of the hydrolysis of neuronal ATP resulting in excessive stimulation of the P2 purinergic receptors and thereby a potential aberration of the purinergic neurotransmission system. This result is in line with previous studies that suggested that alkaloid’s E-NTPDase inhibitory abilities are associated with their neurotoxicity potentials (Colović et al. 2015; Senger et al. 2005). In addition, the present study also observed inhibition of ecto 5-nucleotidase by C. sativa, D. stramonium, Nicotiana tabacum, and Carica papaya alkaloid extracts which coupled with that observed on the ATPase and ADPase activities could result in excess depletion of adenosine levels extracellularly. Meanwhile, excess depletion of adenosine, the neuromodulator implicated in memory and synaptic plasticity formation, has been implicated with adrenergic neurotransmission impairment (Ademiluyi et al. 2016a). Furthermore, scopolamine (i.p.) administration at 1 mg/kg has been observed to reduce ecto 5-nucleotidase activity accompanied by a memory impairment (Marisco et al. 2013). It is therefore hypothesized that the inhibition of the ecto 5-nucleotidase activity observed in the alkaloid extracts in the present study can be compared to that observed in scopolamine administered by Marisco et al. (2013), suggesting that the alkaloid extracts mediated a similar pathway similar to that of scopolamine. This is even consistent with the fact that D. stramonium contains a tangible scopolamine content. Meanwhile, the ability of C. papaya (male and female) to alter the purinergic system activity can be associated with its metal chelating abilities as observed in the present study. This finding is consistent with that observed by Da Silva et al. 2006, stating that ecto-5-nucleotidase/CD73 and ecto-nucleoside triphosphate diphosphohydrolase (E-NTPDase) enzymes depend on divalent metal ions to achieve maximal catalytic activities. Na+/K+-ATPase is the enzyme implicated in the maintenance of electrochemical gradients across cell membranes. Aberration of this equilibrium was observed by de Lores Arnaiz and Ordieres (2014) to cause depolarization of nerve endings, accompanied by the influx of calcium ions into the cells and thereby, the exit of neurotransmitters with resultant neuronal swelling. The present study observed a significant inhibition of the enzyme, Na+/K+-ATPase, and results were of the pattern D. stramonium ˃ C. sativa ˃ N. tabacum ˃ C. papaya (male) ˃ C. papaya (female) in descending order of Na+/K+-ATPase activity inhibition.
Furthermore, the present study observed that incubation of rat brain homogenate with the alkaloid extracts of Datura stramonium caused the highest significant (p < 0.05) elevation of TBARS production ex vivo followed by Cannabis sativa, then Nicotiana tabacum, Carica papaya male, and Carica papaya female respectively. TBARS production has been implicated as a major diagnostic index of lipid peroxidation in neurodegeneration. Lipid peroxidation is the free radical-arbitrated oxidation of polyunsaturated fatty acids that involves chain reactions that ultimately result in deleterious effects on essential biological macromolecules. Studies have it that this oxidative reaction results in lipid hydroperoxide formation which is characterized by further degradation into several aldehydic compounds such as hydroxyalkenals (Panigrahi et al. 2018).
Conclusion
The present study was able to show that alkaloid extracts of Cannabis sativa, Nicotiana tabacum Datura stramonium, and Carica papaya (male and female) were able to modulate the enzyme activities of the monoaminergic, cholinergic, and purinergic systems, and the oxidative stress levels of the rat brain ex vivo in a concentration-dependent manner. The inhibitory effects and elevated lipid peroxidation observed in the present studies by the alkaloid extracts of Datura stramonium, Cannabis sativa, Nicotiana tabacum, and Carica papaya (male and female) might be the underline mechanism by which they elicit psychoactivity as observed in folklore and the neurotoxicity observed in abusers during clinical observations. However, in vivo and clinical studies are recommended to ascertain the neurological and toxicological effects of the alkaloid extracts at the cellular and molecular levels.
References
Adefegha SA, Omojokun OS, Oboh G, Fasakin O, Ogunsuyi O (2015) Modulatory effects of ferulic acid on cadmium-induced brain damage. J Evid Based Complement Altern Med. https://doi.org/10.1177/2156587215621726
Ademiluyi AO, Ogunsuyi OB, Oboh G (2016a) Alkaloid extracts from Jimson weed (Datura stramonium L.) modulate purinergic enzymes in rat brain. Neuro Toxicol 56:107–117. https://doi.org/10.1016/j.neuro.2016.06.012
Ademiluyi AO, Ogunsuyi OB, Oboh G, Agbebi OJ (2016b) Jimson weed (Datura stramonium L.) alkaloid extracts modulate cholinesterase and monoamine oxidase activities in vitro: possible modulatory effect on neuronal function. Comp Clin Pathol 25:733–741. https://doi.org/10.1007/s00580-016-2257-6
Baggio S, Deline S, Studer J, Mohler-Kuo M, Daeppen JB, Gmel G (2014) Routes of administration of cannabis used for nonmedical purposes and associations with patterns of drug use. J Adolesc Health 54(2):235–240. https://doi.org/10.1016/j.jadohealth.2013.08.013
Colović MB, Vasic VM, Avramovic NS, Gajic MM, Djuric DM, Krstic DZ (2015) In vitro evaluation of neurotoxicity potential and oxidative stress responses of diazinon and its degradation products in rat brain synaptosomes. Toxicol Lett 233(1):29–37. https://doi.org/10.1016/j.toxlet.2015.01.003
Cunha-Oliveira T, Rego AC, Oliveira CR (2008) Cellular and molecular mechanisms involved in the neurotoxicity of opioid and psychostimulant drugs. Brain Res Rev 58 (1):192–208
da Silva AC, Balz D, de Souza JDA, Morsch VM, Corrêa MC, Zanetti GD (2006) Inhibition of NTPDase: 50-nucleotidase, Na+/K+-ATPase and acetylcholinesterase activities by subchronic treatment with Casearia sylvestris. Phytomed 13(7):509–514
de LoresArnaiz GR, Ordieres MGL (2014) Brain Na+, K+-ATPase activity in aging and disease. Int J Biomed Sci 10(2):85
Feng L-Y, Battulga A, Han E, Chung H, Li J-H (2017) New psychoactive substances of natural origin: a brief review. J Food Drug Anal 25(3):461–471. https://doi.org/10.1016/j.jfda.2017.04.001
Fiske CH, Subbarow Y (1925) The colorimetric determination of phosphorus. J Biol Chem 66(2):375–400
Panigrahi GK, Verma N, Singh N, Asthana S, Gupta SK, Tripathi A, Das M (2018) Interaction of anthraquinones of Cassia occidentalis seeds with DNA and Glutathione. Toxicol Rep 5:164–172. https://doi.org/10.1016/j.toxrep.2017.12.024
Graziano S, Orsolini L, Rotolo MC, Tittarelli R, Schifano F, Pichini S (2017) Herbal highs: review on psychoactive effects and neuropharmacology. Curr Neuropharmacol 15:750–761. https://doi.org/10.2174/1570159X14666161031144427
Harborne JB (1998) Phytochemical methods: a guide to modern techniques of plant analysis, 3rd edn. Chapman and Hall, London, p 302
Hasin DS, O’Brien CP, Auriacombe M, Borges G, Bucholz K, Budney A, Compton WM, Crowley T, Ling W, Petry NM, Schuckit M, Grant BF (2013) DSM-5 criteria for substance use disorders: recommendations and rationale. Am J Psychiatry 170(8):834–851
Herraiz T, Chaparro C (2005) Human monoamine oxidase is inhibited by tobacco smoke: β-carboline alkaloids act as potent and reversible inhibitors. Biochem Biophys Res Comm 326(2):378–386. https://doi.org/10.1016/j.bbrc.2004.11.033
Heymann D, Reddington M, Kreutzberg GW (1984) Subcellular localization of 5-nucleotidase in rat brain. J Neurochem 43:971–978. https://doi.org/10.1111/j.1471-4159.1984.tb12832.x
Marisco PC, Carvalho FB, Rosa MM, Girardi BA, Gutierres JM, Jaques JA, Salla APS, Pimentel VC, Schetinger MRC, Leal DBR, Mello CF, Rubin MA (2013) Piracetam prevents scopolamine-induced memory impairment and decrease of NTPDase, 5′-nucleotidase and adenosine deaminase activities. Neurochem Res 38:1704–1714. https://doi.org/10.1007/s11064-013-1072-6
Oboh G, Ademosun AO, Ogunsuyi OB, Oyedola ET, Olasehinde TA, Oyeleye SI (2018) In vitro anticholinesterase, antimonoamine oxidase and antioxidant properties of alkaloid extracts from kola nuts (Cola acuminata and Cola nitida). J Complement Integr Med 16:1–12. https://doi.org/10.1515/jcim-2016-0155
Ogunmoyole T, Adeyeye RI, Olatilu BO, Akande OA, Agunbiade OJ (2019) Multiple organ toxicity of Datura stramonium seed extracts. Toxicol Rep 6:983–989. https://doi.org/10.1016/j.toxrep.2019.09.011
Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358
Parle M (2011) Papita fruit: a delicious remedy for depression. Int J Res Ayurveda Pharm 2(4):1358–1364
Perry NS, Houghton PJ, Theobald A, Jenner P, Perry EK (2000) In-vitro inhibition of human erythrocyte acetylcholinesterase by Salvia lavandulaefolia essential oil and constituent terpenes. J Pharm Pharmacol 52(7):895–902. https://doi.org/10.1211/0022357001774598
Rodrigues SR, Caldeira C, Castro BB, Gonçalves F, Nunes B, Antunes SC (2011) Cholinesterase (ChE) inhibition in pumpkinseed (Lepomis gibbosus) as environmental biomarker: ChE characterization and potential neurotoxic effects of xenobiotics. Pestic Biochem Physiol 99(2):181–188. https://doi.org/10.1016/j.pestbp.2010.12.002
Schetinger MRC, Morsch VM, Bonan CD, Wyse AT (2007) NTPDase and 5’- nucleotidase activities in physiological and disease conditions: new perspectives for human health. Biofactors 31(2):77–98. https://doi.org/10.1002/biof.5520310205
Senger MR, Rico EP, de Bem Arizi M, Rosemberg DB, Dias RD, Bogo MR (2005) Carbofuran and Malathion inhibit nucleotide hydrolysis in zebrafish (Danio rerio) brain membranes. Toxicology 212(2):107–115. https://doi.org/10.1016/j.tox.2005.04.007
Turski W, Turska E, Grossbell M (1973) Modification of the spectrophotometric method of the determination of monoamine oxidase. Enzyme 14:211–220
Ujváry I (2014) Psychoactive natural products: overview of recent developments. Ann 1st Super Sanità 50(1):12–27
World Drug Report (2015) Vienna, UN Office on Drugs and Crime, 2015 (https://www.unodc.org/unodc/en/frontpage/2015/June/2015-world-drug-report-finds-drug-use-stable--access-to-drug-and-hiv-treatment-stilllow.html. Accessed 11 Dec 2015)
Wyse AT, Streck EL, Barros SV, Brusque AM, Zugno AL, Wajner M (2000) Methylmalonate administration decreases Na+, K+-ATPase activity in cerebral cortex of rats. Neuroreport 11:2331–2334
Zhi K, Yang Z, Sheng J, Shu Z, Shi Y (2016) A peroxidase-linked spectrophotometric assay for the detection of monoamine oxidase inhibitors. Iran J Pharm Res: IJPR 15(1):131–139
Acknowledgements
All authors of the present study acknowledged the Functional Food and Nutraceutical Unit of the Department of Biochemistry, Federal University of Technology, Akure, for their support in terms of free laboratory, reagent, and equipment usage.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Statement of animal welfare
The care and use of Laboratory Animals were approved by the Federal University of Technology Akure ethical committee, which was followed strictly and in compliance with the National Institute of Health guidelines. Ethical approval was obtained from the Centre for Research and Development (CERAD), Federal University of Technology, Akure, with the number FUTA/ETH/2020/016.
Conflict of interest
The authors declare no competing interests.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Fasakin, O.W., Oboh, G. & Ademosun, A.O. Neuromodulatory evaluation of commonly abused plants ex vivo: a comparative study. Comp Clin Pathol 30, 671–680 (2021). https://doi.org/10.1007/s00580-021-03259-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00580-021-03259-4