Cannabis Health Science Studies Index

A B C D E F G H I J K L M N O P Q R S T U V W X Y Z
 
To utilize this website to your benefit, use any cannabis related key words or phrases including any symptoms, illnesses, or disease. You may also scroll from here.
Loading

ANTIOXIDANT PROPERTIES  & Cannabis studies completed

Overview

An antioxidant is a molecule that inhibits the oxidation of other molecules. Oxidation is a chemical reaction involving the loss of electrons or an increase in oxidation state. Oxidation reactions can produce free radicals. In turn, these radicals can start chain reactions. Antioxidants are found in many foods, including fruits and vegetables, including hemp seed.

See nutrition of hemp seeds
 

Science & Research

1997 - Study- Biological screening of 100 plant extracts for cosmetic use (II): anti-oxidative activity and free radical scavenging activity.

1998 - Study - Cannabidiol and (−)Δ9-tetrahydrocannabinol are neuroprotective antioxidants.

1998 - News ~ Cannabinoid Antioxidant Protects Brain Cells -- Without the High.

2000 - Study- Cannabinoids protect cells from oxidative cell death: a receptor-independent mechanism.

2003 - Patent - US Patent 6630507 - Cannabinoids as antioxidants and neuroprotectants.

2005 - Study ~ Synergistic Interactions between Cannabinoids and Environmental Stress in the Activation of the Central Amygdala.

2005 - Study ~ Protective effects of Δ9-tetrahydrocannabinol against N-methyl-D-aspartate-induced AF5 cell death.

2005 - Study ~ Comparison of Cannabidiol, Antioxidants, and Diuretics in Reversing Binge Ethanol-Induced Neurotoxicity.

2006 - Study ~ In vivo effects of CB1 receptor ligands on lipid peroxidation and antioxidant defense systems in the rat brain of healthy and ethanol-treated rats.

2007- Study ~ Evaluation of the neuroprotective effect of cannabinoids in a rat model of Parkinson's disease: importance of antioxidant and cannabinoid receptor-independent properties.

2007 - Study ~ Repeated Treatment with Cannabidiol but Not Delta9-tetrahydrocannabinol Has a Neuroprotective Effect Without the Development of Tolerance.

2007 - Study ~ Cannabinoids and neuroprotection in motor-related disorders.

2008- Study - Cannabidiol in medicine: a review of its therapeutic potential in CNS disorders.

2008 - Study ~ The role of the endocannabinoid system in Alzheimer's disease: facts and hypotheses.

2009 - Study ~ Cannabidiol Attenuates Cisplatin-Induced Nephrotoxicity by Decreasing Oxidative/Nitrosative Stress, Inflammation, and Cell Death.

2010 - Study ~ Effect of (-)-Delta(9)-tetrahydrocannabinoid on the hepatic redox state of mice.

2010 - Study ~ Acute administration of cannabidiol in vivo suppresses ischaemia-induced cardiac arrhythmias and reduces infarct size when given at reperfusion.

2011 - Study ~ Cannabidiol reduces lipopolysaccharide-induced vascular changes and inflammation in the mouse brain: an intravital microscopy study.

201 2- Study ~ Antioxidant Activities and Oxidative Stabilities of Some Unconventional Oilseeds.

2012 - Study ~ Δ(8) -Tetrahydrocannabivarin prevents hepatic ischaemia/reperfusion injury by decreasing oxidative stress and inflammatory responses through cannabinoid CB(2) receptors.

2012 - Study ~ Cannabidiol treatment ameliorates ischemia/reperfusion renal injury in rats.

2012 - Study ~ Effect of extraction conditions on total polyphenol contents, antioxidant and antimicrobial activities of Cannabis sativa L

2012 - Study ~ The isolation and identification of two compounds with predominant radical scavenging activity in hempseed (seed of Cannabis sativa L.).

2013 - Study ~ Neuroprotective effects of Cannabis sativa leaves extracts on α-Motoneurons density after sciatic nerve injury in rats

2013 - Study ~ Understanding the Molecular Aspects of Tetrahydrocannabinol and Cannabidiol as Antioxidants

2013 - News ~ Marijuana may improve stamina, rejuvenate brain

2013 - News ~ New Study Shows Cannabinoids Improve Efficiency Of Mitochondria And Remove Damaged Brain Cells

2013 - News ~ Marijuana's Memory Paradox

2014 - Study ~ Cannabidiol protects liver from binge alcohol-induced steatosis by mechanisms including inhibition of oxidative stress and increase in autophagy

Cannabidiol and (−)Δ9-tetrahydrocannabinol are neuroprotective antioxidants

The neuroprotective actions of cannabidiol and other cannabinoids were examined in ratcortical neuron cultures exposed to toxic levels of the excitatory neurotransmitter glutamate. Glutamate toxicity was reduced by both cannabidiol, a nonpsychoactive constituent of marijuana, and the psychotropic cannabinoid (−)Δ9-tetrahydrocannabinol (THC).

Cannabinoids protected equally well against neurotoxicity mediated by N-methyl-d-aspartate receptors, 2-amino-3-(4-butyl-3-hydroxyisoxazol-5-yl)propionic acid receptors, or kainate receptors. N-methyl-d-aspartate receptor-induced toxicity has been shown to be calcium dependent; this study demonstrates that 2-amino-3-(4-butyl-3-hydroxyisoxazol-5-yl)propionic acid/kainate receptor-type neurotoxicity is also calcium-dependent, partly mediated by voltage sensitive calcium channels.

The neuroprotection observed with cannabidiol and THC was unaffected by cannabinoid receptor antagonist, indicating it to be cannabinoid receptor independent. Previous studies have shown that glutamate toxicity may be prevented by antioxidants. Cannabidiol, THC and several synthetic cannabinoids all were demonstrated to be antioxidants by cyclic voltametry. Cannabidiol and THC also were shown to prevent

hydroperoxide-induced oxidative damage as well as or better than other antioxidants in a chemical (Fenton reaction) system and neuronal cultures. Cannabidiol was more protective against glutamate neurotoxicity than either ascorbate or α-tocopherol, indicating it to be a potent antioxidant. These data also suggest that the naturally occurring, nonpsychotropic cannabinoid, cannabidiol, may be a potentially useful therapeutic agent for the treatment of oxidative neurological disorders such as cerebral ischemia.

Cannabinoid components of marijuana are known to exert behavioral and psychotropic effects but also to possess therapeutic properties including analgesia, ocular hypotension, and antiemesis. This report examines another potential therapeutic role for cannabinoids as neuroprotectants and describes their mechanism of action in rat cortical neuronal cultures.
 
During an ischemic episode, large quantities of the excitatory neurotransmitter glutamate are released. This event causes neuronal death by over-stimulating N-methyl-d-aspartate receptors (NMDAr) and 2-amino-3-(4-butyl-3-hydroxyisoxazol-5-yl)propionic acid (AMPA) and kainate-type receptors and results in metabolic stress and accumulation of toxic levels of intracellular calcium.
In vitro and in vivo studies  have demonstrated that such neurotoxicity can be reduced by antioxidants or antagonists to NMDAr and AMPA/kainate receptors.
 
Antioxidants such as α-tocopherol are effective neuroprotectants because of their ability to reduce the toxic reactive oxygen species (ROS) formed during ischemic metabolism. Cannabinoids like (−)Δ9-tetrahydrocannabinol (THC) and its psychoactive analogues also have been reported to be neuroprotective against glutamate toxicity in vitro.
 
Cannabinoids have been suggested to prevent glutamate neurotoxicity by activating cannabinoid receptors, which can reduce calcium influx through voltage sensitive calcium channels. A synthetic cannabinoid (HU-211) also has been demonstrated to be neuroprotective even though it does not activate cannabinoid receptors. This compound is an atypical cannabinoid, however, in that it, unlike other cannabinoids, directly antagonizes NMDAr and possesses some antioxidant properties.
 
The present study examines classical cannabinoids as neuroprotectants in vitro but focuses on the nonpsychoactive cannabinoid cannabidiol. Like THC, cannabidiol is a natural component of the marijuana plant, Cannabis sativa, although unlike THC, cannabidiol does not activate cannabinoid receptors and so is devoid of psychoactive effects.
 
This study reports that cannabidiol and other cannabinoids such as THC are potent antioxidants that protect neurons from glutamate-induced death without cannabinoid receptor activation.

MATERIALS AND METHODS
Materials.
Cannabidiol, THC, and reagents other than those specifically listed below were purchased from Sigma. Cyclothiazide, glutamatergic ligands, and MK-801 were obtained from Tocris Cookson (Bristol, U.K.). Dihydrorhodamine was supplied by Molecular Probes. Tert-butyl hydroperoxide, tetraethylammonium chloride, ferric citrate, and sodium dithionite were all purchased from Aldrich. Agatoxin and conotoxin were obtained through Biomol (Plymouth Meeting, PA). All culture media were GIBCO/BRL products.
 
Solution Preparation.
Solutions of cannabinoids, cyclothiazide, and other lipophiles were prepared by evaporating a 10 mM ethanolic solution (under a stream of nitrogen) in a siliconized microcentrifuge tube. Dimethyl sulfoxide (<0.05% of final volume) was added to ethanol to prevent the lipophile from completely drying onto the tube wall.
 
After evaporation, 1 ml of culture media was added, and the drug was dispersed by using a high power sonic probe. Special attention was used to ensure the solution did not overheat or generate foam. After dispersal, all solutions were made to their final volume in siliconized glass tubes by mixing with an appropriate quantity of culture media.
 
Neuronal Cultures.
Primary cortical neuron cultures were prepared according to the method of Ventra et al. In brief, fetuses were extracted by C-section from a 17-day pregnant Wistar rat, and the fetal brains were placed into phosphate buffered saline. The cortices then were dissected out, cut into small pieces, and incubated with papain for 9 min at 37°.
 
After this time, the tissue was dissociated by passage through a fire-polished Pasteur pipette, and the resultant cell suspension was separated by centrifugation over a gradient consisting of 10 mg/ml BSA and 10 mg/ml ovomucoid (a trypsin inhibitor) in Earle’s balanced salt solution.
 
The pellet then was resuspended in high glucose, phenol red-free DMEM containing 10% fetal bovine serum, 2 mM glutamine, 100 units of penicillin, and 100 μg/ml streptomycin (DMEM). Cells were counted, were tested for vitality by using the trypan blue exclusion test, and were seeded onto poly-d-lysine coated 24 multiwell plates. After 96 hr, 10 μM fluorodeoxyuridine and 10 μM uridine were added to block glial cell growth. This protocol results in a highly neuron-enriched culture.
 
Preparation of (Type I) Astrocytes and Conditioned Media.
Astrocyte-conditioned DMEM (phenol red-free) was used throughout the AMPA/kainate toxicity procedure and after glutamate exposure in the NMDAr-mediated toxicity protocol. Media were conditioned by 24 hr of treatment over a confluent layer of type I astrocytes prepared from 2-day-old Wistar rat pups.
 
In brief, cortices were dissected, were cut into small pieces, were digested enzymatically with 0.25% trypsin, and then were dissociated mechanically by passage through a plastic pipette. The cell suspension then was plated into untreated 75-cm2 T-flasks, and, after 24 hr, the media were replaced and unattached cells were removed. Once astrocytes achieved confluency, cells were divided into four flasks. Media for experiments were conditioned by a 24-hr exposure to these astrocytes, after which time they were frozen at −20°C until use. Astrocyte cultures were used to condition DMEM for no longer than 2 months.
 
NMDAr-Mediated Toxicity Procedure.
NMDAr-mediated glutamate toxicity was examined by exposing neurons (cultured for 14–18 days) to 250 μM glutamate for 10 min in a phenol red-free and magnesium-free saline. The saline was composed of 125 mM NaCl, 25 mM Glucose, 10 mM Hepes (pH 7.4), 5 mM KCl, 1.8 mM calcium chloride, and 5% BSA. After exposure, cells were washed twice with saline and were incubated for 18 hr in conditioned DMEM. Toxicity was prevented completely by addition of the NMDAr antagonist MK-801 (500 nM) (data not shown).
 
AMPA and Kainate Receptor-Mediated Toxicity Procedures.
Unlike NMDAr, which are regulated by magnesium ions, AMPA/kainate receptors rapidly desensitize after ligand binding. To examine AMPA and kainate receptor-mediated toxicity, neurons were cultured for 7–13 days and then were exposed to 100 μM glutamate and 50 μM cyclothiazide (used to prevent AMPA receptor desensitization).
 
Cells were incubated with glutamate in the presence of 500 nM MK-801 for 18–20 hr before analysis. Specific AMPA and kainate receptor ligands also were used to separately examine the effects of cannabinoids on AMPA and kainate receptor-mediated events. Fluorowillardiine (1.5 μM) and 4-methyl glutamate (10 μM) were used to investigate AMPA and kainate receptor-mediated toxicity, respectively. When specifically examining kainate receptor activity, cyclothiazide was replaced with 0.15 mg/ml Concanavalin-A.
 
Although the neuron preparation technique described above results in a largely neuronal culture, a minority of astrocytic cells remain. Astrocytes are highly resistant to glutamate toxicity because of their lack of functional NMDAr, although glutamate toxicity in astrocytes has been observed to prevent AMPA receptor desensitization if cyclothiazide is present.
 
To examine whether AMPA/kainate-type toxicity affects astrocytes in our cultures, astrocytes (as prepared above) were exposed to glutamate under the same conditions used on neuron-enriched cultures. Under these conditions, astrocytes were resistant to glutamate toxicity, with 20 hr of exposure resulting in a lactate dehydrogenase release of only 5% above background, compared with 100–200% of the background observed in neuron-enriched cultures (data not shown). It was concluded, therefore, that astrocyte contamination does not contribute substantially to the effects of glutamate in our neuronal cultures.
 
ROS Toxicity Assay.
To examine the effects of cannabinoids on ROS toxicity, 7- to 13-day-old cultured neurons were incubated with 300 μM tert-butyl hydroperoxide (an oxidant) in conditioned DMEM. Tert-butyl hydroperoxide was used because its miscibility with both water and lipids allows oxidation to occur in both cytosolic and membrane-delimited cellular compartments.
 
Toxicity Assay.
Cell toxicity was assessed 18–20 hrs after insult by measuring lactate dehydrogenase release into the (phenol red-free) culture media. Experiments were conducted with triple or quadruple values at each point, and all plates contained positive (glutamate alone) and baseline controls. The assay was validated by comparison with a tetrazolium-based viability assay (XTT). Results were similar with either system, although lactate dehydrogenase release was used in this study because it provided a greater signal to noise ratio than the XTT assay.
 
Cyclic Voltametry.
Cyclic voltametry was performed with an EG & G Princeton Applied Research potentiostat/galvanostat (model 273/par 270 software). The working electrode was a glassy carbon disk with a platinum counter electrode and silver/silver chloride reference. Tetraethylammonium chloride in acetonitrile (0.1 M) was used as an electrolyte. Cyclic voltametry scans were done from 0 to +1.8 V at scan rate of 100 mV per second.
 
Iron-Catalysed Dihydrorhodamine Oxidation (Fenton Reaction).
The antioxidant activities of each of the compounds were evaluated by their ability to prevent oxidation of dihydrorhodamine to the fluorescent compound rhodamine.
 
Oxidant was generated by ferrous catalysis (diothionite-reduced ferric citrate) of tert-butyl hydroperoxide in a 50:50 water-to-acetonitrile (vol/vol) solution. Dihydrorhodamine (50 μM) was incubated with 300 μM tert-butyl hydroperoxide and 0.5 μM iron for 5 min. After this time, oxidation was assessed by spectrofluorimetry (Excitation = 500 nm, Emission = 570 nm). Various concentrations of cannabinoids and butylhydroxytoluene (BHT) were included to examine their ability to prevent dihydrorhodamine oxidation.
 
Data Analysis.
Data are reported as mean values plus and minus standard error. Significance was examined by using a Student’s t test, (P ≤ 0.05). Kinetic data was analyzed by using GraphPad’s prism software package (GraphPad, San Diego) for PC.

RESULTS
Cannabidiol Blocks NMDAr and AMPA and Kainate Receptor-Mediated Neurotoxicity.
Glutamate neurotoxicity can be mediated by NMDAr, AMPA receptors, or kainate receptors. To examine NMDAr-mediated toxicity, rat cortical neurons were exposed to glutamate for 10 min in a magnesium-free medium, and the level of lactate dehydrogenase released was used as an index of cell injury. To examine AMPA/kainate receptor-mediated toxicity, neurons were incubated for 20 hr with glutamate or a specific AMPA or kainate receptor ligand (fluorowillardiine or 4-methyl-glutamate, respectively). An NMDAr antagonist (MK-801) and an agent to prevent receptor desensitization also were included. Cannabidiol prevented cell death equally well (EC50 of 2–4 μM) in both NMDAr and AMPA/kainate toxicity models (Fig. (Fig.11 A and B).
 
Similar data also was observed when glutamate was replaced with either AMPA-specific or kainate receptor-specific ligands (data not shown). These results suggest that cannabidiol protects similarly, regardless of whether toxicity is mediated by NMDA, AMPA, or kainate receptors.
 
Figure 1
Figure 1
Effect of cannabidiol on NMDAr- (A) and AMPA/kainate receptor- (B) mediated neurotoxicity. Data shown represents mean values ± SEM from a single experiment with four replicates. Each experiment was repeated on at least four occasions with (more ...)
 
AMPA/Kainate Toxicity Is Calcium Dependent.
Increased calcium influx is known to be a key factor in NMDAr-induced cell death, but its role in AMPA and kainate toxicity is less clear. It has been suggested that AMPA/kainate receptors may not directly allow entry of sufficient calcium to kill cells. However, AMPA/kainate receptors flux large amounts of sodium, which can depolarize cell membranes. Such depolarization may activate both voltage-sensitive calcium channels and facilitate NMDAr activation .
 
In this way, AMPA/kainate receptor stimulation may lead indirectly to accumulation of toxic intracellular calcium levels. Addition of the calcium chelator EDTA reduced toxicity in a concentration-dependent manner, demonstrating the involvement of calcium in AMPA/kainate-type toxicity (data not shown). EDTA (2 mM) eliminated ≈70% of glutamate toxicity (Fig. (Fig.2)2) even though the presence of MK-801 prevented NMDAr activation.
 
Toxicity also was reduced by inhibitors to L-, N-, and P/Q- type calcium channels (nifedipine, agatoxin IVa, and conotoxin GVIa, respectively; Fig. Fig.2),2), indicating that voltage-sensitive calcium channels also are involved in AMPA/kainate-type toxicity. However, a combination of these calcium channel inhibitors did not completely block EDTA-preventable (calcium-dependant) cell death.
 
Figure 2
Figure 2
The involvement of calcium and calcium channels in AMPA/kainate-mediated toxicity. The effects of 2 mM EDTA and various combinations of the voltage-sensitive calcium channel inhibitors ω-Agatoxin IVa (Ag) (250 nM), ω-Conotoxin (more ...)
 
Neuroprotection by Tetrahydrocannabinol.
Unlike cannabidiol, THC is a ligand for the brain cannabinoid receptor, and this action has been proposed to explain the ability of THC to protect neurons from NMDAr toxicity in vitro. However, in AMPA/kainate receptor toxicity assays, THC and cannabidiol were similarly protective, suggesting that cannabinoid neuroprotection may be independent of cannabinoid receptor activation. This was confirmed by inclusion of a cannabinoid receptor antagonist, SR-141716A (Fig. (Fig.3).3). Neither THC or cannabidiol neuroprotection was affected by cannabinoid receptor antagonist.
 
Figure 3
Figure 3
Effect of THC, cannabidiol, and cannabinoid receptor antagonist on glutamate induced neurotoxicity. Neurons exposed to glutamate in an AMPA/kainate receptor toxicity model were incubated with 10 μM cannabidiol or THC in the presence or (more ...)
 
Cannabinoids as Antioxidants.
Cells use easily oxidizable compounds such as glutathione, ascorbate, and α-tocopherol as antioxidants that protect important cellular structures (e.g., DNA, proteins, and membranes) from ROS damage. Studies have suggested that ROS damage may be involved in glutamate neurotoxicity.
 
To investigate whether cannabinoids could protect neurons against glutamate by reacting with ROS, the antioxidant properties of cannabidiol and other cannabinoids were assessed. Cyclic voltametry, a procedure that measures the ability of a compound to accept or donate electrons under a variable voltage potential, was used to measure the oxidation potentials of several natural and synthetic cannabinoids. Cannabidiol, THC, and the synthetic cannabinoid HU-211 all donated electrons at a similar potential as the antioxidant BHT. Anandamide (arachidonyl-ethanolamide), which is not a cannabinoid in structure but is an endogenous ligand for the cannabinoid receptor, did not undergo oxidation in this assay (Fig. (Fig.44A).
 
Three other cannabinoids, cannabinol, nabilone, and levanantrodol, also were tested, and they, too, exhibited oxidation profiles similar to cannabidiol and THC (data not shown).
 
Figure 4
Figure 4
(A) A comparison of the oxidation potentials of cannabinoids and the antioxidant BHT. The oxidation profiles of (750 μM) BHT, cannabinoids, and anandamide were compared by cyclic voltametry. Anandamide, a cannabinoid receptor ligand with a noncannabinoid (more ...)
 
The ability of cannabinoids to be oxidized readily suggests that they may possess antioxidant properties comparable to BHT. These properties were examined further in a Fenton reaction (iron-catalyzed ROS generation). Tert-butyl hydroperoxide was used to generate ROS and oxidize dihydrorhodamine into the fluorescent compound rhodamine. Cannabidiol, THC, and BHT all prevented dihydrorhodamine oxidation in a similar, concentration-dependent manner (Fig. (Fig.44B), indicating cannabinoids to be comparable to BHT in antioxidant potency.
 
To confirm that cannabinoids act as antioxidants in the intact cell, neurons were incubated with tert-butyl hydroperoxide and varying concentrations of cannabidiol (Fig. (Fig.55A). The oxidant was chosen for its solubility in both aqueous and organic solvents, thereby facilitating oxidation in both cytosolic and membrane cell compartments. As observed in studies with glutamate, cannabidiol protected neurons against ROS toxicity in a concentration-related manner. Cannabidiol also was compared with antioxidants in an AMPA/kainate toxicity protocol. Neurons were exposed to glutamate and equimolar cannabidiol, α-tocopherol, BHT, or ascorbate (Fig. (Fig.55B). Although all of the antioxidants attenuated glutamate toxicity, cannabidiol was significantly more protective than either α-tocopherol or ascorbate.
 
Figure 5
Figure 5
(A) The effect of cannabidiol on oxidative toxicity in neuronal cultures. Tert-butyl hydroperoxide-induced toxicity was examined in the presence or absence of cannabidiol. (B) Comparison of antioxidants and cannabidiol for their ability to prevent glutamate (more ...)

DISCUSSION
The nonpsychoactive marijuana constituent cannabidiol was found to prevent both glutamate neurotoxicity and ROS-induced cell death. The psychoactive principle of Cannabis, THC, also blocked glutamate neurotoxicity with a similar potency to cannabidiol. In both cases, neuroprotection was unaffected by cannabinoid receptor antagonist. This suggests that cannabinoids may have potentially useful therapeutic effects that are independent of psychoactivity-inducing cannabinoid receptors and so are not necessarily accompanied by psychotropic side effects.
 
Cannabidiol blocked glutamate toxicity in cortical neurons with equal potency regardless of whether the insult was mediated by NMDAr, AMPA receptors, or kainate receptors. This suggests that either cannabinoids antagonize all three glutamate receptors with the same affinity, or, more likely, their site of action is downstream of initial receptor activation events.
 
Neurotoxic concentrations of glutamate induce massive calcium influx through NMDAr that ultimately kills the cell. This study has demonstrated that the toxic effects of glutamate are also calcium-dependent when mediated by AMPA/kainate receptors. Both EDTA (a calcium chelator) and voltage-sensitive calcium channel inhibitors reduced AMPA-/kainate-type neurotoxicity, indicating that a portion of calcium influx-associated AMPA-/kainate-receptor activation is mediated by secondary activation of calcium channels.
 
However, the mixture of calcium channel inhibitors and NMDAr antagonist did not eliminate completely glutamate toxicity or reduce cell death as efficiently as EDTA. This suggests that although toxicity resulting from AMPA/kainate receptor stimulation may be caused by calcium entering the cell by several routes, it is not caused exclusively by calcium channel activity. These studies also demonstrate that NMDAr activation is not required for AMPA-/kainate-type toxicity [as suggested.
 
Accumulation of ROS has been shown to be involved in NMDAr-mediated cell death. The current study has similarly demonstrated that AMPA/kainate receptor-induced toxicity also involves ROS formation and may be prevented with antioxidant treatment. Cannabidiol and THC were found to be comparable with BHT (antioxidant) in both their ability to prevent dihydrorhodamine oxidation (Fenton reaction) and their cyclic voltametric profiles.
 
Synthetic cannabinoids such as HU-211, nabilone, and levanantradol also exhibited similar profiles. Anandamide, which is a natural cannabinoid receptor ligand but is not structurally related to cannabinoids, did not give an antioxidant-like profile by cyclic voltametry, which indicates that cannabinoids can act as reducing agents (in a chemical system). To confirm that cannabinoids also can function as antioxidants in living cells, a lipid hydroperoxide was used to generate ROS toxicity in neuronal cultures. As observed in the Fenton reaction system, cannabidiol attenuated this ROS-induced neurotoxicity. These observations indicate that many cannabinoids exert a considerable protective antioxidant effect in neuronal cultures.
 
The similarity of the voltamagrams observed with cannabidiol, HU-211, and several other cannabinoids also suggests that the reported antioxidant effect of HU-211 is not a feature unique to this atypical cannabinoid, (as previously implied; e.g., ref. 11) but, rather, a common property of classical cannabinoid structures.
 
The potency of cannabidiol as an antioxidant was examined by comparing it on an equimolar basis with other commonly used antioxidants. Cannabidiol protected neurons to a greater degree than either of the dietary antioxidants, α-tocopherol or ascorbate. As in the Fenton reaction system, cannabidiol protected neurons with comparable efficacy to the potent antioxidant BHT. The similar antioxidant abilities of cannabidiol and BHT in this chemical system and their comparable protection in neuronal cultures implies that cannabidiol neuroprotection is caused by an antioxidant effect.
 
The antioxidative properties of cannabinoids suggest a therapeutic use as neuroprotective agents, and the particular properties of cannabidiol make it a good candidate for such development. Although cannabidiol was similar in neuroprotective capacity to BHT, cannabidiol has no known tumor-promoting effects [unlike BHT ].
 
The lack of psychoactivity associated with cannabidiol allows it to be administered in higher doses than would be possible with psychotropic cannabinoids such as THC. Furthermore, the ability of cannabidiol to protect against neuronal injury without inhibiting NMDAr may reduce the occurrence of toxicity or side effects associated with NMDAr antagonists. Previous studies have indicated that cannabidiol is not toxic, even when chronically administered to humans or given in large acute doses [700 mg/day.
 
In vivo studies to examine the efficacy of cannabidiol as a treatment for experimentally induced ischemic stroke are currently in progress.
ABBREVIATIONS
 
AMPA2-amino-3-(4-butyl-3-hydroxyisoxazol-5-yl)propionic acid
 
BHTbutylhydroxytoluene
 
NMDArN-methyl-d-aspartate receptors
 
ROSreactive oxygen species
 
THC(−)Δ9-tetrahydrocannabinol


References
1. Welch S P, Stevens D L. J Pharmacol ExpTher. 1992;262:8–10.
2. Merritt J C, Crawford W J, Alexander P C, Anduze A L, Gelbart S S. Ophthalmology. 1980;87:222–228. [PubMed]
3. Abrahamov A, Mechoulam R. Life Sci. 1995;56:2097–2102. [PubMed]
4. Choi D W, Koh J Y, Peters S. J Neurosci. 1988;8:185–196. [PubMed]
5. Ciani E, Groneng L, Voltattorni M, Rolseth V, Contestabile A, Paulsen R E. Brain Res. 1996;728:1–6. [PubMed]
6. MacGregor D G, Higgins M J, Jones P A, Maxwell W L, Watson M W, Graham D I, Stone T W. Brain Res. 1996;727:133–144. [PubMed]
7. Skaper S D, Buriani A, Dal Toso R, Petrell I L, Romanello L, Facci L, Leon A. Proc Natl Acad Sci USA. 1996;93:3984–3989. [PMC free article] [PubMed]
8. Hampson A J, Bornheim L M, Scanziani M, Yost C S, Gray A T, Hansen B M, Leonoudakis D J, Bickler P E. J Neurochem. 1998;70:671–676. [PubMed]
9. Twitchell W, Brown S, Mackie K. J Neurophysiol. 1997;78:43–50. [PubMed]
10. Biegon A. Ann NY Acad Sci. 1995;765:314. [PubMed]
11. Eshhar N, Striem S, Kohen R, Tirosh O, Biegon A. Eur J Pharmacol. 1995;283(1–3):19–29. [PubMed]
12. Mansbach R S, Rovetti C C, Winston E N, Lowe J A., III Psychopharmacol. 1996;124:315–22. [PubMed]
13. Ventra C, Porcellini A, Feliciello A R, Gallo A, Paolillo M, Mele E, Avedimento V E, Schettini G. J Neurochem. 1996;66:1752–1761. [PubMed]
14. Grimaldi M, Pozzoli G, Navarra P, Preziosi P, Schettini G. J Neurochem. 1994;63:344–350. [PubMed]
15. Zhou L M, Gu Z Q, Costa A M, Yamada K A, Mansson P E, Giordano T, Skolnick P, Jones K A. J Pharmacol Exp Ther. 1997;280:422–427. [PubMed]
16. Amin N, Pearce B. Neurochem Int. 1997;30:271–276. [PubMed]
17. Holopainen I, Saransaari P, Oja S S. Neurochem Res. 1994;19:111–115. [PubMed]
18. Van Bockstaele E J, Colago E E J. J Comp Neurol. 1996;369:483–496. [PubMed]
19. David J C, Yamada K A, Bagwe M R, Goldberg M P. J Neurosci. 1996;16:200–209. [PubMed]
20. Roehm N W, Rodgers G H, Hatfield S M, Glasebrook A L. J Immunol Methods. 1991;142:257–265. [PubMed]
21. Hack N, Balazs R. J Neurochem. 1995;65:1077–1084. [PubMed]
22. Berman F W, Murray T F J. J Biochem Toxicol. 1996;11:111–119. [PubMed]
23. Deupree D L, Tang X W, Yarom M, Dickman E, Kirch R D, Schloss J V, Wu J Y. Neurochem Int. 1996;29:255–261. [PubMed]
24. Devane W A, Dysarz F A, Johnson M R, Melvin L S, Howlett A C. Mol Pharmacol. 1988;34:605–613. [PubMed]
25. Thompson J A, Bolton J L, Malkinson A M. Exp Lung Res. 1991;17:439–453. [PubMed]
26. Lindenschmidt R C, Tryka A F, Goad M E, Witschi H P. Toxicology. 1986;38:151–160. [PubMed]
27. Auer R N. Psychopharmacol Bull. 1994;30:585–591. [PubMed]
28. Cunha J M, Carlini E A, Pereira A E, Ramos O L, Pimentel C, Gagliardi R, Sanvito W L, Lander N, Mechoulam R. Pharmacology. 1980;21:175–185. [PubMed]
29. Consroe P, Laguna J, Allender J, Snider S, Stern L, Sandyk R, Kennedy K, Schram K. Pharmacol Biochem Behav. 1991;40:701–708. [PubMed]

Biological screening of 100 plant extracts for cosmetic use (II): anti-oxidative activity and free radical scavenging activity

Kim BJ, Kim JH, Kim HP, Heo MY 
Biological screening of 100 plant extracts for cosmetic use (II): anti-oxidative activity and free radical scavenging activity. [Journal Article]
Int J Cosmet Sci 1997 Dec; 19(6):299-307.

Methanol aqueous extracts of 100 plants were screened for anti-oxidative activity using Fenton's reagent/ethyl linoleate system and for free radical scavenging activity using the 1,1-diphenyl-2-picryl hydrazyl free radical generating system.

The results suggest that 14 plants - Alpinia officinarum, Areca catechu, Brassica alba, Cannabis sativa, Curcuma longa, Curcuma aromatica, Eugenia caryophyllata, Evodia officinalis, Paeonia suffruticosa, Rhaphanus sativus, Rheum palmatum, Rhus verniciflua, Trapa bispinosa, Zanthoxylum piperitum - may be potential sources of anti-oxidants. Eight plants - Citrus aurantium, Cornus officinalis, Gleditsia japonica, Lindera strychnifolia, Phragmites communis, Prunus mume, Schizandra chinensis, Terminalia chebula - may be the potential source of free radical scavengers from natural plant.

 

 

Top    Home


 

Cannabinoids protect cells from oxidative cell death: a receptor-independent mechanism

J Pharmacol Exp Ther. 2000 Jun;293(3):807-12.

Chen Y, Buck J.

Department of Pharmacology, Joan & Sanford I. Weill Medical College of Cornell University, New York, NY 10021, USA.

Abstract

Serum is required for the survival and growth of most animal cells. In serum-free medium, B lymphoblastoid cells and fibroblasts die after 2 days.

 We report that submicromolar concentrations of Delta(9)-tetrahydrocannabinol (THC), Delta(8)-THC, cannabinol, or cannabidiol, but not WIN 55,212-2, prevented serum-deprived cell death. Delta(9)-THC also synergized with platelet-derived growth factor in activating resting NIH 3T3 fibroblasts.

The cannabinoids' growth supportive effect did not correlate with their ability to bind to known cannabinoid receptors and showed no stereoselectivity, suggesting a nonreceptor-mediated pathway. Direct measurement of oxidative stress revealed that cannabinoids prevented serum-deprived cell death by antioxidation.

The antioxidative property of cannabinoids was confirmed by their ability to antagonize oxidative stress and consequent cell death induced by the retinoid anhydroretinol. Therefore, cannabinoids act as antioxidants to modulate cell survival and

 

 

Top    Home


 

Cannabidiol in medicine: a review of its therapeutic potential in CNS disorders

Scuderi C, Filippis DD, Iuvone T, Blasio A, Steardo A, Esposito G 
Cannabidiol in medicine: a review of its therapeutic potential in CNS disorders. [JOURNAL ARTICLE]
Phytother Res 2008 Oct 9.


Cannabidiol (CBD) is the main non-psychotropic component of the glandular hairs of Cannabis sativa. It displays a plethora of actions including anticonvulsive, sedative, hypnotic, antipsychotic, antiinflammatory and neuroprotective properties.

However, it is well established that CBD produces its biological effects without exerting significant intrinsic activity upon cannabinoid receptors. For this reason, CBD lacks the unwanted psychotropic effects characteristic of marijuana derivatives, so representing one of the bioactive constituents of Cannabis sativa with the highest potential for therapeutic use.

The present review reports the pharmacological profile of CBD and summarizes results from preclinical and clinical studies utilizing CBD, alone or in combination with other phytocannabinoids, for the treatment of a number of CNS disorders. Copyright (c) 2008 John Wiley & Sons, Ltd.

 

 

 

Top    Home


 

US Patent 6630507 - Cannabinoids as antioxidants and neuroprotectants

Abstract

Cannabinoids have been found to have antioxidant properties, unrelated to NMDA receptor antagonism. This new found property makes cannabinoids useful in the treatment and prophylaxis of wide variety of oxidation associated diseases, such as ischemic, age-related, inflammatory and autoimmune diseases.

The cannabinoids are found to have particular application as neuroprotectants, for example in limiting neurological damage following ischemic insults, such as stroke and trauma, or in the treatment of neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease and HIV dementia.

Nonpsychoactive cannabinoids, such as cannabidoil, are particularly advantageous to use because they avoid toxicity that is encountered with psychoactive cannabinoids at high doses useful in the method of the present invention. A particular disclosed class of cannabinoids useful as neuroprotective antioxidants is formula (I) wherein the R group is independently selected from the group consisting of H, CH3, and COCH3. ##STR1##

Patent References

2304669

(3S-4S)-7-hydroxy-Ɗ6 -tetrahydrocannabinols

Method for the production of 6,12-dihydro-6-hydroxy-cannabidiol and the use thereof for the production of trans-delta-9-tetrahydrocannabinol

NMDA-blocking pharmaceutical compositions

Neuroprotective pharmaceutical compositions of 4-phenylpinene derivatives and certain novel 4-phenylpinene compounds
Patent #: 5434295
Issued on: 07/18/1995
Inventor: Mechoulam, et al.

Nitroxides as protectors against oxidative stress

Method of inhibiting oxidants using alkylaryl polyether alcohol polymers

NMDA-blocking pharmaceuticals

Certain tetrahydrocannabinol-7-oic acid derivatives

(3S,4S)-delta-6-tetrahydrocannabinol-7-oic acids and derivatives thereof, processors for their preparation and pharmaceutical compositions containing them

Other References

  • Windholz et al., The Merck Index, Tenth Edition (1983) p. 241, abstract No. 1723.
  • Mechoulam et al., "A Total Synthesis of d1-Ɗ1 -Tetrahydrocannabinol, the Active Constituent of Hashish1," Journal of the American Chemical Society, 87:14:3273-3275 (1965)
  • Mechoulam et al., "Chemical Basis of Hashish Activity," Science, 18:611-612 (1970)
  • Ottersen et al., "The Crystal and Molecular Structure of Cannabidiol," Acta Chem. Scand. B 31, 9:807-812 (1977)
  • Cunha et al., "Chronic Administration of Cannabidiol to Healthy Volunteers and Epileptic Patients1," Pharmacology, 21:175-185 (1980)
  • Consroe et al., "Acute and Chronic Antiepileptic Drug Effects in Audiogenic Seizure-Susceptible Rats," Experimental Neurology, Academic Press Inc., 70:626-637 (1980)
  • Turkanis et al., "Electrophysiologic Properties of the Cannabinoids," J. Clin. Pharmacol., 21:449S-463S (1981)
  • Carlini et al., "Hypnotic and Antielpileptic Effects of Cannabidiol," J. Clin. Pharmacol., 21:417S-427S (1981)
  • Karler et al., "The Cannabinoids as Potential Antiepileptics," J. Clin. Pharmacol., 21:437S-448S (1981)
  • Consroe et al., "Antiepileptic Potential of Cannabidiol Analgos," J. Clin. Pharmacol., 21:428S-436S (1981)
  • Colasanti et al., "Ocular Hypotension, Ocular Toxicity,a nd Neurotoxicity in Response to Marihuana Extract and Cannabidiol," Gen Pharm., Pergamon Press Ltd., 15(6):479-484 (1984)
  • Colasanti et al., "Intraocular Pressure, Ocular Toxicity and Neurotoxicity after Administration of Cannabinol or Cannabigerol," Exp. Eye Res., Academic Press Inc., 39:251-259 (1984)
  • Volfe et al., "Cannabinoids Block Release of Serotonin from Platelets Induced by Plasma frm Migraine Patients," Int. J. Clin. Pharm. Res., Bioscience Ediprint Inc., 4:243-246 (1985)
  • Agurell et al., "Pharmacokinetics and Metabolism of Ɗ1 -Tetrahydrocannabinol and Other Cannabinoids with Emphasis on Man*," Pharmacological Reviews, 38(1):21-43 (1986)
  • Karler et al., "Different Cannabinoids Exhibit Different Pharmacological and Toxicological Properties,"NIDA Res. Monogr., 79:96-107 (1987)
  • Samara et al., "Pharmacokinetics of Cannabidiol in Dogs," Drug Metabolism and Disposition, 16(3):469-472 (1988)
  • Choi, "Glutamate Neurotoxicity and Diseases of the Nervous System," Neuron, Cell Press, 1:623-634 (1988)
  • Eshhar et al., "Neuroprotective and Antioxidant Activities of HU-211, A Novel NMDA Receptor Antagonist," European Journal of Pharmacology, 283:19-29 (1995)
  • Skaper et al., "The ALIAmide Palmitoylethanolamide and Cannabinoids, but not Anandamide, are Protective in a Delayed Postglutamate Paradigm of Excitotoxic Death in Cerebellar Granule Neurons," Neurobiology, Proc. Natl. Acad. Sci. USA, 93:3984-3989 (1996)
  • Alonso et al., "Simple Synthesis of 5-Substituted Resorcinols: A Revisited Family of Interesting Bioactive Molecules," J. Org. Chem., American Chemical Society, 62(2):417-421 (1997)
  • Combes et al. "A Simple Synthesis of the Natural 2,5-Dialkylresorcinol Free Radical Scavenger Antioxidant: Resorstation," Synthetic Communications, Marcel Dekker, Inc., 27(21):3769-3778 (1997)
  • Shohami et al., "Oxidative Stress in Closed-Head Injury: Brain Antioxidant Capacity as an Indicator of Functional Outcome," Journal of Cerebral Blood Flow and Metabolism, Lippincott-Raven Publishers, 17(10):1007-1019 (1997)
  • Zurier et al., "Dimethylheptyl-THC-11 OIC Acid," Arthritis & Rheumatism, 41(1):163-170 (1998)
  • Hampson et al., "Dual Effects of Anandamide on NMDA Receptor-Mediated Responses and Neurotransmission," Journal of Neurochemistry, Lippincott-Raven Publishers, 70(2):671-676 (1998)
  • Hampson et al., "Cannabidiol and (-)Ɗ9 -tetrahydrocannabiono are Neuroprotective Antioxidants," Medical Sciences, Proc. Natl. Acad. Sci. USA, 8268-8273 (1998)


Top    Home