ArticlePDF Available

Cannabidiol lowers incidence of diabetes in non-obese diabetic mice

Authors:

Abstract and Figures

Cannabidinoids are components of the Cannabis sativa (marijuana) plant that have been shown capable of suppressing inflammation and various aspects of cell-mediated immunity. Cannabidiol (CBD), a non-psychoactive cannabidinoid has been previously shown by us to suppress cell-mediated autoimmune joint destruction in an animal model of rheumatoid arthritis. We now report that CBD treatment significantly reduces the incidence of diabetes in NOD mice from an incidence of 86% in non-treated control mice to an incidence of 30% in CBD-treated mice. CBD treatment also resulted in the significant reduction of plasma levels of the pro-inflammatory cytokines, IFN-gamma and TNF-alpha. Th1-associated cytokine production of in vitro activated T-cells and peritoneal macrophages was also significantly reduced in CBD-treated mice, whereas production of the Th2-associated cytokines, IL-4 and IL-10, was increased when compared to untreated control mice. Histological examination of the pancreatic islets of CBD-treated mice revealed significantly reduced insulitis. Our results indicate that CBD can inhibit and delay destructive insulitis and inflammatory Th1-associated cytokine production in NOD mice resulting in a decreased incidence of diabetes possibly through an immunomodulatory mechanism shifting the immune response from Th1 to Th2 dominance.
Content may be subject to copyright.
Cannabidiol lowers incidence of diabetes in non-obese diabetic mice
L. WEISS
1
, M. ZEIRA
1
, S. REICH
1
, M. HAR-NOY
1
, R. MECHOULAM
2
, S. SLAVIN
1
,&
R. GALLILY
3
1
Department of Bone Marrow Transplantation & Cancer Immunotherapy, Hadassah University Hospital, POB 12000,
Jerusalem 91120, Israel,
2
Department of Medical Chemistry and Natural Products, Hebrew University Medical Faculty,
Jerusalem 91120, Israel, and
3
Department of Immunology, Hebrew University Medical Faculty, Jerusalem 91120, Israel
(Received 25 January 2005; in revised form 14 September 2005)
Abstract
Cannabidinoids are components of the Cannabis sativa (marijuana) plant that have been shown capable of suppressing
inflammation and various aspects of cell-mediated immunity. Cannabidiol (CBD), a non-psychoactive cannabidinoid has
been previously shown by us to suppress cell-mediated autoimmune joint destruction in an animal model of rheumatoid
arthritis. We now report that CBD treatment significantly reduces the incidence of diabetes in NOD mice from an incidence of
86% in non-treated control mice to an incidence of 30% in CBD-treated mice. CBD treatment also resulted in the significant
reduction of plasma levels of the pro-inflammatory cytokines, IFN-g and TNF-a. Th1-associated cytokine production of in
vitro activated T-cells and peritoneal macrophages was also significantly reduced in CBD-treated mice, whereas production of
the Th2-associated cytokines, IL-4 and IL-10, was increased when compared to untreated control mice. Histological
examination of the pancreatic islets of CBD-treated mice revealed significantly reduced insulitis. Our results indicate that
CBD can inhibit and delay destructive insulitis and inflammatory Th1-associated cytokine production in NOD mice resulting
in a decreased incidence of diabetes possibly through an immunomodulatory mechanism shifting the immune response from
Th1 to Th2 dominance.
Keywords: Type 1 diabetes, cannabidiol, Th1/Th2 biology, IFN-g
Introduction
The Cannabis sativa (marijuana) plant contains
components that are termed cannabinoids. Cannabi-
noids are known to have a variety of potential
therapeutic effects which include analgesic, anti-
inflammatory and immunosuppressive properties
[1,2]. The major psychoactive cannabinoid is Delta
9-tetrahydrocannabinol (THC) whose effects are
mediated through the CB1 and the CB2 subtypes of
cannabinoid receptors found in the brain and
lymphoid tissues [3]. THC has potent immunosup-
pressive properties and can modulate Th1/Th2
balance by enhancing Th2 and suppressing Th1
immune responses [47]. However, the psychoactive
effects of THC limit its consideration as a therapy for
Th1-mediated autoimmune diseases.
Cannabidiol (CBD), on the other hand, is a
cannabinoid that has potential for clinical research and
therapeutic use because it does not produce psychoac-
tive effects due to a low affinity for the CB1 and CB2
receptors [8,9] and is also well tolerated without side
effects when chronically administered to humans
[10,11]. However, the anti-inflammatory, immunosup-
pressive and Th1/Th2 altering effects of CBD have not
been as well characterized as those of THC.
CBD has previously been shown to be better than
THC in inhibiting pro-inflammatory IL-1, TNF-a,
and IFN-g cytokine release by peripheral blood
ISSN 0891-6934 print/ISSN 1607-842X online q 2006 Taylor & Francis
DOI: 10.1080/08916930500356674
Correspondence: S. Slavin, Department of Bone Marrow Transplantation & Cancer Immunotherapy, Hadassah University Hospital, POB
12000, Jerusalem 91120, Israel. Tel: 972 2 677 7270. Fax: 972 2 642 2731. E-mail: slavin@hadassah.org.il
Autoimmunity, March 2006; 39(2): 143–151
mononuclear cells [12]. We previously demonstrated
that CBD was effective in suppressing progression of
autoimmune joint destruction in the collagen-induced
arthritis animal model of rheumatoid arthritis, a
Th1-mediated disease [13]. The anti-autoimmune
effects of CBD were associated with reduction in
synovial cell TNF-a production, inhibition of reactive
oxygen release from zymosan-stimulated neutrophils,
suppression of macrophage nitric oxide (NO) pro-
duction and suppression of joint-specific T-cell
proliferation and IFN-g production.
CBD has also recently been shown to have potent
anti-inflammatory effects in rat paw carrageen-
induced inflammation [14] associated with reduction
in prostaglandin E2 (PGE2), tissue cyclooxygenase
(COX) activity, and production of oxygen-derived free
radicals and production of NO.
Since CBD has been shown to inhibit production of
IL-1b, TNF-a and IFN-g, as well as the inflammatory
mediators NO, COX and PGE2 and these factors are
known to be involved in the pathway of autoimmune
islet cell destruction leading to diabetes, we decided to
test the anti-autoimmune effects of CBD in the NOD
mouse model of type 1 diabetes.
The aim of this study was to determine if CBD
could prevent or delay diabetes occurrence in the
spontaneous NOD mouse model of type 1 diabetes.
We examined the effect of CBD on the development of
diabetes in untreated and treated female NOD mice
by assaying glucose levels in urine and analyzing
insulitis and beta cell integrity in histopathological
sections of the pancreases.
We report that CBD significantly inhibited insulitis,
beta cell destruction and the occurrence of overt
diabetes in the NOD mice. In addition, we found that
CBD treatment resulted in suppressed production of
the Th1-associated cytokines, IL-12, IFN-g and
TNF-a, and enhanced production of the Th2-
associated cytokines, IL-4 and IL-10, suggesting a
possible deviation from destructive Th1 immunity to
protective Th2 immunity.
Material and methods
Mice
Female non-obese diabetic NOD/LtJ mice, 6 12-
week-old, were obtained from Harlan, Israel. Mice
were fed standard laboratory animal chow ad libit um
and were kept in a specific pathogen free (SPF)
animal housing facility. The study was conducted in
compliance with international laws on animal
experimentation and “Principles of Laboratory
Animal Care” (NIH publication No. 86 23, revised
1985) were followed. The study was approved by the
Ethical Committee of the Hebrew University Medical
School.
CBD
CBD was extracted from cannabis resin (hashish) as
previously reported [15]. For in vivo injection, CBD
was first dissolved in ethanol and then Cremophor EL
(Sigma) was added at a 1:1 ratio. This solution was
then further diluted in saline so that the final solution
was ethanol/Cremophor/saline (1:1:18).
Experimental design
CBD was administered i.p. at a dose of 5 mg/kg/day.
Ten to twenty injections (5 times a week) were
administered to 612-week-old female NOD mice.
Two groups of mice served as controls, one group
received the Cremophor/ethanol/saline vehicle only
and the other group remained untreated. The level of
urine glucose was assayed once or twice a week by test
strips (Medi-Test Combi 9 Macherey-Nagel, Duren)
and was considered positive after the appearance of
glucosuria in at least two consecutive determinations.
Peritoneal macrophage culture
Peritoneal lavages from each untreated and CBD-
treated mouse were centrifuged and the peritoneal
macrophage pellets were resuspended and plated in 24-
well plates for 2 h. The wells were washed to remove
non-adherent cells and fresh DMEM with 10% FCS
supplemented with 1 mg/ml LPS (W.E.Coli type 055:B5
Difco-Bacto Labs, Detroit, MI, USA) was added to
wells for further over night incubation. Harvested
supernatants were centrifuged to remove any cells and
stored at 2 808C until assayed for cytokines.
Splenic lymphocyte culture
Spleen cells from each untreated and CBD-treated
mouse were washed in PBS, resuspended in RPMI
medium supplemented with 10% Fetal Bovine Serum,
100 U/ml penicillin, 100 mg/ml streptomycin and
2 mM L-glutamine and dispensed into 24-well plates
(Greiner Labs, Germany) at 2 £ 10
6
cells/ml medium
per well. Cells were stimulated with 2.5 mg/ml ConA
(Sigma, St. Louis MI, USA) and cultured at 378Cina
5% CO
2
humidified incubator. Supernatants were
harvested after 48 h incubations, cleared by centrifu-
gation and stored at 2 808C until assayed for cytokines.
Cytokine assay
Levels of cytokines in the plasma and in the
supernatants of splenic lymphocytes and peritoneal
macrophages of the mice were assayed by “sandwich”
ELISA technique. ELISA reagents were purchased as
Opt. EIA Cytokine ELISA sets from BD Biosciences
(San Diego, CA, USA) and were used according to the
manufacturer’s protocol as previously described [16].
L. Weiss et al.144
Histopathology
Pancreatic tissue was fixed in 4% buffered formalin
and was embedded in paraffin, and 5-mm sections
were stained with hematoxylin and eosin. Sections
containing a total of more than 17 islets from each
pancreas were screened and scored by two indepen-
dent observers and were graded according to the
following scores: intact islets; partially infiltrated islets
and peri-infiltrated islets; totally infiltrated islets; and
destroyed islets.
Results
CBD reduces diabetes disease incidence in NOD mice
To determine the ability of CBD to suppress the
cumulative incidence or delay the onset of diabetes in
NOD mice, we chose to treat female animals that
already had insulitis, but did not have overt disease.
This status was chosen to approximate the immuno-
logical status of a pre-type 1 diabetic human patient.
In NOD mice, insulitis begins at 45 weeks of age and
is present in 100% of females [17,18]. Therefore, we
used only female mice that were a minimum of 6
weeks old.
Disease onset in female NOD mice had been
reported to begin at 8 10 weeks of age with a
cumulative frequency of 70 90% [17,18]. However,
many differences exist between NOD mouse colonies
[19]. In our colony, the female NOD mice developed
overt diabetes at a median of 14 weeks, therefore,
CBD treatments were administered to mice that were
up to 12 weeks of age. All mice were normoglycemic at
the time of initiation of experiments.
Female NOD mice 612 weeks of age were treated
with 10 20 i.p. injections of 5 mg/kg CBD. There was
no difference in the treatment outcome as a function
of the number of CBD injections, therefore, the results
of all experiments were pooled for statistical analysis.
These results are summarized in Figure 1 and Table I.
We show that 86% of the untreated control mice
developed diabetes in a median of 14 weeks (range of
1020 weeks). The CBD-treated mice demonstrated
a significant reduction in disease incidence compared
to the untreated mice with only 30% of CBD-treated
mice developing disease ( p ¼ 0.00003). Further, the
fraction of mice that developed disease in the CBD-
treated group (6 of 20 mice) had a delayed onset of
disease and this delay was significant when compared
to the untreated control group (20 week median onset
in the CBD-treated group compared to 14 week
median onset in the control group, p ¼ 0.0001).
In another experiment, CBD-treated mice were
observed for diabetes incidence at 26 weeks. The
control mice (5 mice) all developed diabetes at a
median of 17 weeks (range 1520 weeks) while 3 of 5
(60%) CBD-treated mice remained diabetes-free at
26 weeks.
CBD is a hydrophobic molecule and poorly soluble
in water, therefore, it was solubilized in Cremophor
EL (a polyethoxylated castor oil) and ethanol for
injection (vehicle). To determine if the vehicle would
have an effect independent of CBD, we evaluated a
group of mice treated with vehicle alone. Vehicle-
treated mice had a disease incidence of 69%, which
was lower, but not significantly different than, the
disease incidence in the untreated mice. However, the
time to disease onset in the fraction of vehicle-treated
mice that developed disease was significantly extended
compared to the time of onset in untreated mice
(median of 17.5 weeks in vehicle-treated mice
compared to a median of 14 weeks in control
mice, p ¼ 0.05).
The incidence of disease in the CBD-treated group
(30%) was significantly lower than both the untreated
control (86%, p ¼ 0.00003) and the vehicle-treated
control (69%, p ¼ 0.009). Time to the onset of disease
in the fraction of mice that developed diabetes in the
CBD-treated mice (median 20 weeks) was also
significantly extended when compared to both the
time to onset in the untreated control group (median
14 weeks) and the vehicle-treated group (17.5 weeks).
These data demonstrate the ability of CBD
treatment to suppress the cumulative incidence and
delay the onset of diabetes in NOD mice.
Plasma levels of IFN-
g
and TNF-
a
are reduced in CBD-
treated mice
NOD mice have high levels of IFN-g and TNF-a
secretion [20] and increased IFN-g levels are
correlated with islet destruction and development of
diabetes [21]. We, therefore, determined whether CBD
had an effect on plasma levels of these inflammatory
Figure 1. Incidence of diabetes in NOD mice treated with 5 mg/kg
CBD i.p., vehicle control and untreated mice. The incidence of
diabetes in the CBD treated group is significantly less when
compared to both the control and vehicle groups.
Cannabidiol lowers incidence of diabetes 145
cytokines. Both cytokine levels were significantly
reduced in the CBD-treated mice (Table II).
We found that IFN-g levels in plasma of treated
mice were 72% lower than the levels in the vehicle
treated mice ( p , 0.01) and 79% lower than the levels
of untreated diabetic mice ( p , 0.01). The levels of
TNF-a in the plasma of treated mice were 77% lower
than the vehicle treated and untreated control groups
( p , 0.05). These results demonstrate a suppression
of inflammatory cytokine levels in the plasma of CBD-
treated mice compared to untreated and vehicle-
treated control groups.
Shift in Th1/Th2 cytokine balance in activated splenocytes
and peritoneal macrophages
Diabetes is correlated with an altered balance of
Th1/Th2 cytokines with an excess of Th1 cytokines
and lack of Th2 cytokines [2224]. We, therefore,
investigated the effect of CBD on Th1 (IFN-g, IL-12
and TNF-a) and Th2 (IL-4 and IL-10) cytokine
production in in vitro activated splenocytes and
peritoneal macrophages. Analysis of these responses
revealed a general shift towards production of Th2
cytokines in CBD-treated mice.
Splenocytes from each of the CBD-treated and
vehicle-treated (control) NOD mice were polyclonally
activated by cultivation for 48 h with ConA. Levels of
IL-4 and IFN-g produced by the splenocytes were
assayed. The results are shown in Table III. We found
a profound (99%) suppression of IFN-g in CBD-
treated mice, while the amount of IL-4 production
was increased by 41%. These effects were statistically
significant ( p , 0.05).
The levels of IL-10, IL-12 and TNF-a cytokines
produced by LPS-activated peritoneal macrophages
were also modulated by CBD treatment of NOD mice.
The decrease in the CBD treated mice of the
pro-inflammatory cytokines IL-12 (by 74%) and
TNF-a (by 77%) was significant compared to the
control ( p , 0.05), whereas IL-10, which inhibits the
expressionoftheabovepro-inflammatorycytokineswas
remarkably increased by greater than 5-fold ( p , 0.05)
in macrophages from CBD treated mice. The levels of
IL-10 were also elevated in unstimulated spleen cell
supernatants from CBD-treated mice (10 pg/ml IL-10
in the CBD-treated group compared to 0.5 pg/ml in the
vehicle control).
Overall, these results indicate that CBD therapy
induced a Th2-enriched environment in the periphery
of NOD-mice.
CBD therapy prevents insulitis development
To test the therapeutic effect of CBD on the
progression of insulitis, tissue sections of CBD-treated
mice and control groups (3 mice from each group)
were examined for the presence of infiltrating cells
Table I. Occurrence of diabetes in CBD-treated NOD mice.
Treatment
No. of mice with
overt diabetes
Median time to diabetes
incidence (range) in weeks p value*
Total no. of mice
in group % Diabetic mice p value
Untreated 19 14 (1020) 0.05 22 86
Vehicle 9 17.5 (1420) 0.0001‡ 13 69 N.S.
CBD 6 20 (1520) 0.008{ 20 30 P , 0.05
§
Cumulative appearance of diabetes in female NOD mice treated with 5 mg/kg CBD.
* Significance of delay in disease onset evaluated by Kaplan-Meier log bank Statistics.
CBD-treated vs. untreated.
{
CBD-treated vs vehicle.
N.S. ¼ Non-significant.
Significance of disease incidence evaluated by Student’s t-test.
§
Significant when CBD treated group compared to both untreated and vehicle-treated.
Table II. Suppression of IFN-g and TNF-a levels in plasma of CBD-treated NOD mice.
IFN-g (pg/ml) TNF-a
Average ^SE Average ^ SE
Treatment of NOD mice 13 weeks 17 weeks 20 weeks
CBD 43 ^ 10* 10 ^ 4* 6.7 ^ 0.88†
Vehicle 94 ^ 21 68 ^ 12 29 ^ 14.1
Untreated diabetic 100 ^ 22 118 ^ 42 N.D.
IFN-g levels in plasma of female NOD mice untreated and treated with CBD at 13 (n ¼ 5) and 17 (n ¼ 10) weeks of age correspondingly.
TNF-a levels in plasma of CBD treated and vehicle treated mice at 20 weeks of age (n ¼ 3). N.D. ¼ not done.
* Significant difference p , 0.01 of CBD vs vehicle and vs untreated control by the Kaplan-Meier statistics at both 13 weeks and 17 weeks.
Differences between experimental and control values and between experimental and vehicle were analyzed for significance ( p # 0.05) by
one-tail distribution-free Mann Whitney U-test.
L. Weiss et al.146
at 20 weeks of age. Histological examination revealed
a reduction in the degree and severity of insulitis in
CBD-treated mice compared to controls (Table IV).
Representative islets from each group are shown in
Figure 2.
Intact and partially infiltrated islets were observed
in 87% of the CBD-treated mice, whereas only 4%
and 15% were detected in the untreated and vehicle-
treated control mice, respectively. Only one islet out
of 54 analyzed was fully destroyed in CBD-treated
mice, compared to 21% and 51% of the islets in the
vehicle-treated and untreated control mice,
respectively.
These results suggest that CBD-treatment inhibited
the progression to destructive insulitis. This data
together with the cytokine data is indicative of a Th2
response that suppressed the beta cell destruction in
the pancreas, since peri-insulitis has previously been
associated with Th2 responses [25,26].
Discussion
CBD is a non-psychoactive component of marijuana
that possesses anti-inflammatory and immunosup-
pressive properties. Here we demonstrate the ability of
CBD to significantly reduce the occurrence and delay
the onset of overt diabetes in female NOD mice. NOD
mice present with a clinical and immunological course
very similar to human type 1 diabetes [27,28].
Therefore, these results raise the question of whether
CBD could have a role in prevention of human type 1
diabetes.
The answer to this question will likely depend upon
the further elucidation of the mechanism(s) of the
anti-autoimmune properties of CBD. Specifically, the
determination of whether CBD is acting as a non-
specific immunosuppressive agent or, alternatively (or
in addition) acting through a mechanism of Th1/Th2
immune deviation. The present study suggests that
CBD treatment may function by stimulating a
protective Th2 immune response in NOD mice.
This is demonstrated by a significant reduction in
diabetes incidence in CBD-treated mice with a shift in
regulatory cytokine production in the periphery from
Th1-to Th2-biased.
Non-specific immunosuppressive activity would not
be as desirable a mechanism for diabetes prevention as
would be an immunomodulatory mechanism. Many
non-specific immunosuppressive drugs have been
shown to prevent diabetes in NOD mice [29], but
such therapies have not been as successful in the clinic.
The immunosuppressive drug cyclosporine, for
example, has been administered to children with
new-onset diabetes [30]. While cyclosporine-treated
patients were shown to be more likely to be in
remission after one year than non-treated controls,
this difference was no longer significant at 2 years
[31]. The limited efficacy of this approach combined
Table III. In vivo treatment with CBD modulates cytokine secretion by lymphocytes and peritoneal macrophages.
In vivo treatment of NOD mice Number of mice Cell type+in vitro activation method Cytokine evaluated pg/ml ^ SE Percent change (CBD vs. control)
CBD 7 Sp þ Con A IL-4 30.8 ^ 2.12 þ 41
control 5 Sp þ Con A IL-4 21.8 ^ 2.61
CBD 4 Sp þ Con A IFN-g , 10 2 99
control 4 Sp þ Con A IFN-g 1009 ^ 572
CBD 5 PMB þ LPS IL-10 340 ^ 157.8 þ 530
control 3 PMB þ LPS IL-10 54 ^ 19.98
CBD 5 PMB þ LPS IL-12 25.6 ^ 7.42 2 74
control 3 PMB þ LPS IL-12 98 ^ 45
CBD 10 PMB þ LPS TNF-a 32.7 ^ 2.34 2 78
control 12 PMB þ LPS TNF-a 145 ^ 24.9
Cytokine levels in the supernatants of activated spleen cells (Sp) and peritoneal macrophages (PMB) taken from CBD treated and untreated mice at 20 weeks of age. Differences between CBD and
control values were analyzed for significance ( p # 0.05) by one-tail distribution-free MannWhitney U-test. Differences in all groups were significant.
Cannabidiol lowers incidence of diabetes 147
with safety concerns have limited the acceptance of
cyclosporine therapy as a preventive treatment for
diabetes.
A mechanism of non-specific immunosuppression
requires lifelong treatment which poses substantial
potential risks, including possible enhanced suscepti-
bility to lethal opportunistic infection and increased
risk of malignancy. While the long-term consequences
of diabetes include blindness, renal failure and
increased susceptibility to heart disease, these
complications can be reduced with intensive insulin
therapy aimed at normalizing glucose levels [32].
Therefore, the long-term safety of CBD as a non-
specific immunosuppressive therapy for diabetes
would have to be carefully evaluated prior to use in
humans and before such a therapy could be accepted
as a preventive therapy for diabetes.
On the other hand, if CBD is acting as an
immunomodulating therapy causing the deviation of
the islet-specific immune response from Th1 to Th2,
this might enable CBD to be used to prevent diabetes
in early on-set patients prior to complete beta cell
destruction or even possibly in high risk individuals.
Once the autoimmune response is deviated from a
destructive Th1 response to a protective Th2
response, presumably no additional CBD therapy
would be necessary to maintain tolerance, therefore,
reducing the concern regarding long-term safety
issues.
The evidence presented herein is suggestive of an
immunomodulatory effect of CBD on the clinical
course of diabetes induction in NOD mice, possibly
through an alteration of macrophage function.
Cytokines from macrophages and other innate
Table IV. Inhibition of insulitis by CBD.
Treatment No. of scored islets Intact islets Partially infiltrated islets/peri-insulitis Totally infiltrated islets Fully destroyed islets
CBD 54 26 21 6 1
Vehicle 52 0 2 29 21
Untreated 73 4 7 11 51
Histological analysis of pancreas tissue (fixed in 4% buffered formalin) from mice treated with CBD, vehicle control and untreated control.
Islets were scored according to the degree of mononuclear cell infiltration and islet integrity.
Figure 2. Representative histology of Hematoxylin-Eosin stained pancreases from (A) untreated control mice, mag. x 800, (B) vehicle-
treated mice, mag. x 800, and (C) CBD-treated mice, mag. x 200.
L. Weiss et al.148
immune cells have a critical immunomodulatory role
in skewing the adaptive immune response to either
Th1 or Th2 [33]. In NOD mice, macrophages are
critical for establishing Th1 autoimmunity, as
depletion of macrophages leads to a shift from a Th1
to a Th2 immune response and a reduction in islet cell
destruction [34]. Administration of IL-12, a macro-
phage derived cytokine, to macrophage-depleted
NOD mice, on the other hand, initiates disease [34].
Therefore, CBD may have an immunomodulating
effect through the suppression of macrophage inflam-
matory cytokine production.
Here we demonstrate that CBD significantly
reduced macrophage IL-12 production and that
plasma IFN-g levels were also significantly reduced.
IL-12 is a key cytokine for skewing the development
of adaptive immune responses. IL-12 induces the
differentiation of Th1 cells and the production of
IFN-g [35]. IFN-g in turn potentates the production
of IL-12 [36]. Th1 cytokines, such as IFN-g, also
activate cytotoxic T-cells and stimulate macrophages
to produce pro-inflammatory cytokines (i.e. TNF-a,
IL-1, and IL-6) and nitrogen free radicals (e.g. NO)
which all correlate with islet beta cell destruction
[37]. Therefore, CBD suppression of macrophage
IL-12 production could inhibit Th1-mediated auto-
immunity.
We previously showed that CBD could block
progressive autoimmune arthritis and that treated
mice had significantly suppressed TNF-a production
[13]. Here we again show that CBD caused a
significant reduction in TNF-a production. Anti-
TNF therapy is widely used for treatment of
autoimmune diseases such as rheumatoid arthritis,
Crohn’s disease, ankylosing spondylitis, and psoriatic
arthritis. TNF-a levels are elevated in the serum of
diabetic patients [38] and anti-TNF therapy can
prevent diabetes in NOD mice [39]. Therefore, this is
another potential therapeutic mechanism for CBD in
the clinic.
The alterations in regulatory cytokines in CBD-
treated mice also had an effect on regulatory cytokine
production from T-cells. Activated splenocytes from
the CBD-treated NOD mice were found to produce
dramatically reduced levels of IFN-g and increased
levels of IL-4. These results indicate a shift in the
regulatory cytokine environment of the treated mice
from an environment supportive of development of
Th1 immunity to an environment supportive of Th2
immunity. However, it is not clear whether the
alterations in cytokine production we demonstrate is
the mechanism of disease suppression or is manifested
as a consequence of the suppression.
The increased IL-4 levels and reduced IFN-g levels
in CBD-treated mice would enable the steering of an
islet-specific immune response to Th2. It has been
proposed that type 1 diabetes is the consequence of a
progressive destruction of pancreatic beta cells
mediated by an imbalance between effector Th1 and
regulatory Th2 cell function [40], where Th1
immunity plays a critical role in the development of
diabetes and where Th2 immunity may be protective
[41,42].
Here we show that cells from CBD-treated mice
produced in vitro enhanced IL-4 and IL-10 cytokine
production and such cytokine alterations correlated
with disease suppression. Several reports support that
immunological interventions, which enhance Th2
immune responses in NOD mice are protective
against disease. For example, treatment of NOD
mice with IL-4, a Th2 cytokine, prevents overt
diabetes [4345] and local expression of IL-4 in
pancreatic beta cells in transgenic mice inhibits
insulitis [46]. IL-10, a Th2 associated cytokine, is
correlated with protection [46] and IL-4 and IL-10,
together inhibit diabetes by suppressing Th1 cytokine
production in islet grafts [47].
While our data shows that the regulatory cytokine
environment was altered in CBD-treated mice, this
study did not address the question of whether a
Th1/Th2 shift occurred in the islet antigen-specific
adaptive immune response, and if such a shift
occurred, whether this shift provides long-term
protection from disease without further intervention.
This additional information will be critical in
determining the immunomodulatory role of CBD
and will be the subject of further investigation.
It is possible that the inevitable autoimmune
response to islet antigens in NOD mice could be
skewed to Th2 bias under the influence of a local
cytokine milieu enhanced in IL-4 and reduced in IFN-
g and IL-12 as we demonstrate occurs after treatment
with CBD. It is also possible that an islet-specific
immune response developing under Th2-steering
conditions could lead to a protective Th2 insulitis,
rather than a destructive Th1 insulitis leading to
diabetes [48]. In support of this possible mechanism,
previous studies have demonstrated that an established
insulin-specific autoimmune response in NOD mice
that is highly Th1-biased [49] can be diverted with
immunotherapy to a dominant Th2 immune response
[50]. This suggests that an immunomodulatory
therapy might be able to prevent diabetes through a
mechanism of Th1/Th2 deviation even after the
establishment of disease.
Most patients at onset of diabetes have approxi-
mately 10% of beta cells remaining and these islets
remain functional as evidenced by residual c-peptide
secretion. Approximately 50% of patients diagnosed
with type 1 diabetes will enter a “honeymoon”
remission phase of diabetes and remain insulin
independent within the first year of diagnosis.
Assuming these patients have sufficient residual beta
cells to maintain normoglycemia and insulin indepen-
dence, they will be candidates for immunomodulation
therapy.
Cannabidiol lowers incidence of diabetes 149
CBD is non-psychoactive and has anti-inflamma-
tory and anti-autoimmune properties. The results of
the present study provide further evidence of the
anti-autoimmune effects of CBD. The data presented
herein suggests that CBD-treatment inhibits diabetes
by induction of regulatory Th2 responses. This
possible immunomodulatory effect of CBD is an
interesting focus for further investigation. Confir-
mation of such a mechanism may lead to the clinical
application of this agent in the prevention of type 1
diabetes, as well as possibly in other Th1-mediated
autoimmune diseases.
Acknowledgements
We wish to thank the Danny Cunniff Leukemia
Research Laboratory; the Gabrielle Rich Leukemia
Research Foundation; the Cancer Treatment Research
Foundation; the Novotny Trust; the Szydlowsky
Foundation; the Figure Tree Foundation; Ronne &
Donald Hess; and the Silverstein Family for their
continuous support of our ongoing basic and clinical
research.
References
[1] Mbvundula EC, Rainsford KD, Bunning RA. Cannabinoids in
pain and inflammation. Inflammopharmacology 2004;12:
99114.
[2] Klein TW, Newton C, Friedman H. Cannabinoid receptors
and immunity. Immunol Today 1998;19:373381.
[3] Howlett AC. The cannabinoid receptors. Prostaglandins Other
Lipid Mediat 2002;68-69:619631.
[4] Newton CA, Klein TW, Friedman H. Secondary immunity to
Legionella pneumophila and Th1 activity are suppressed by
delta-9-tetrahydrocannabinol injection. Infect Immun
1994;62:4015 4020.
[5] Klein TW, Newton CA, Nakachi N, Friedman H. Delta 9-
tetrahydrocannabinol treatment suppresses immunity and
early IFN-g, IL-12, and IL-12 receptor beta 2 responses to
Legionella pneumophila infection. J Immunol 2000;
164:6461 6466.
[6] Zhu LX, Sharma S, Stolina M, et al. Delta-9-tetrahydrocan-
nabinol inhibits antitumor immunity by a CB2 receptor-
mediated, cytokine-dependent pathway. J Immunol 2000;
165:373380.
[7] Yuan M, Kiertscher SM, Cheng Q, Zoumalan R, Tashkin DP,
Roth MD. Delta 9-Tetrahydrocannabinol regulates Th1/Th2
cytokine balance in activated human T-cells. J Neuroimmunol
2002;133:124 131.
[8] Munro S, Thomas KL, Abu-Shaar M. Molecular character-
ization of a peripheral receptor for cannabinoids. Nature
1993;365:61 65.
[9] Thomas BF, Gilliam AF, Burch DF, Roche MJ, Seltzman HH.
Comparative receptor binding analyses of cannabinoid
agonists and antagonists. J Pharmacol Exp Ther 1998;
285:285292.
[10] Cunha JM, Carlini EA, Pereira AE, et al. Chronic
administration of cannabidiol to healthy volunteers and
epileptic patients. Pharmacology 1980;21:175185.
[11] Consroe P, Laguna J, Allender J, et al. Controlled clinical trial
of cannabidiol in Huntington’s disease. Pharmacol Biochem
Behav 1991;40:701708.
[12] Watzl B, Scuderi P, Watson RR. Influence of marijuana
components (THC and CBD) on human mononuclear
cell cytokine secretion in vitro.AdvExpMedBiol
1991;288:6370.
[13] Malfait AM, Gallily R, Sumariwalla PF, et al. The
nonpsychoactive cannabis constituent cannabidiol is an oral
anti-arthritic therapeutic in murine collagen-induced arthritis.
Proc Natl Acad Sci USA 2000;97:95619566.
[14] Costa B, Colleoni M, Conti S, et al. Oral anti-inflammatory
activity of cannabidiol, a non-psychoactive constituent of
cannabis, in acute carrageenan-induced inflammation in the
rat paw. Naunyn Schmiedebergs Arch Pharmacol
2004;369:294299.
[15] Gaoni Y, Mechoulam R. The isolation and structure of delta-
1-tetrahydrocannabinol and other neutral cannabinoids from
hashish. J Am Chem Soc 1971;93:217 224.
[16] Ji YH, Weiss L, Zeira M, et al. Allogeneic cell-mediated
immunotherapy of leukemia with immune donor lymphocytes
to upregulate antitumor effects and downregulate antihost
responses. Bone Marrow Transplant 2003;32:495504.
[17] Kikutani H, Makino S. The murine autoimmune diabetes
model: NOD and related strains. Adv Immunol 1992;
51:285322.
[18] Wicker LS, Todd JA, Peterson LB. Genetic control of
autoimmune diabetes in the NOD mouse. Annu Rev Immunol
1995;13:179200.
[19] Pozzilli P, Signore A, Williams AJ, Beales PE. NOD mouse
colonies around the world-recent facts and figures. Immunol
Today 1993;14:193196.
[20] Yaacob NS, Kaderi MA, Norazmi MN. The expression of
cytokine genes in the peritoneal macrophages and splenic
CD4- and CD8-positive lymphocytes of the nonobese diabetic
mice. J Clin Immunol 2004;24:177184.
[21] Rabinovitch A, Suarez-Pinzon WL, Sorensen O, Bleackley
RC, Power RF. IFN-g gene expression in pancreatic islet-
infiltrating mononuclear cells correlates with autoimmune
diabetes in nonobese diabetic mice. J Immunol 1995;154:
48744882.
[22] Almawi WY, Tamim H, Azar ST. Clinical review 103: T helper
type 1 and 2 cytokines mediate the onset and progression of
type I (insulin-dependent) diabetes. J Clin Endocrinol Metab
1999;84:14971502.
[23] Satoh J, Takahashi K, Nakazawa T, Sakata Y, Toyota T.
[Current concept of the pathogenesis of autoimmune type 1
diabetes mellitus]. Nippon Rinsho 1999;57:528533.
[24] Chang JC, Linarelli LG, Laxer JA, et al. Insulin-secretory-
granule specific Tcell clones in human IDDM. J Autoimmun
1995;8:221234.
[25] Gallichan WS, Balasa B, Davies JD, Sarvetnick N. Pancreatic
IL-4 expression results in islet-reactive Th2 cells that inhibit
diabetogenic lymphocytes in the nonobese diabetic mouse.
J Immunol 1999;163:16961703.
[26] Healey D, Ozegbe P, Arden S, Chandler P, Hutton J, Cooke A.
In vivo activity and in vitro specificity of CD4+Th1 and Th2
cells derived from the spleens of diabetic NOD mice. J Clin
Invest 1995;95:2979 2985.
[27] Bach JF, Boitard C. Experimental models of type-I diabetes.
Pathol Immunopathol Res 1986;5:384415.
[28] Bach JF. The Non Obese Diabetic (NOD) mouse, as a model
of Tcell mediated autoimmune disease. C R Acad Sci III
1992;314:4546.
[29] Greiner DL, Rossini AA, Mordes JP. Translating data from
animal models into methods for preventing human auto-
immune diabetes mellitus: Caveat emptor and primum non
nocere. Clin Immunol 2001;100:134 143.
[30] Stiller CR, Dupre J, Gent M, et al. Effects of cyclosporine
immunosuppression in insulin-dependent diabetes mellitus of
recent onset. Science 1984;223:1362 1367.
L. Weiss et al.150
[31] Bougneres PF, Landais P, Boisson C, et al. Limited duration of
remission of insulin dependency in children with recent overt
type I diabetes treated with low-dose cyclosporin. Diabetes
1990;39:12641272.
[32] The effect of intensive treatment of diabetes on the
development and progression of long-term complications in
insulin-dependent diabetes mellitus. The Diabetes Control
and Complications Trial Research Group. N Engl J Med
1993;329:977986.
[33] Swain SL. T-cell subsets. Who does the polarizing? Curr Biol
1995;5:849851.
[34] Jun HS, Santamaria P, Lim HW, Zhang ML, Yoon JW.
Absolute requirement of macrophages for the development
and activation of beta-cell cytotoxic CD8+T-cells in T-cell
receptor transgenic NOD mice. Diabetes 1999;48:3442.
[35] Trinchieri G. Interleukin-12 and the regulation of innate
resistance and adaptive immunity. Nat Rev Immunol
2003;3:133146.
[36] Skeen MJ, Miller MA, Shinnick TM, Ziegler HK. Regulation
of murine macrophage IL-12 production. Activation of
macrophages in vivo, restimulation in vitro, and modulation
by other cytokines. J Immunol 1996;156:11961206.
[37] Ohno Y, Aoki N, Nishimura A. In vitro production of
interleukin-1, interleukin-6, and tumor necrosis factor-alpha
in insulin-dependent diabetes mellitus. J Clin Endocrinol
Metab 1993;77:10721077.
[38] Abdel Aziz MT, Fouad HH, Mohsen GA, Mansour M, Abdel
Ghaffar S. TNF-a and homocysteine levels in type 1 diabetes
mellitus. East Mediterr Health J 2001;7:679688.
[39] Hunger RE, Carnaud C, Garcia I, Vassalli P, Mueller C.
Prevention of autoimmune diabetes mellitus in NOD mice by
transgenic expression of soluble tumor necrosis factor receptor
p55. Eur J Immunol 1997;27:255261.
[40] Rabinovitch A, Suarez-Pinzon WL. Cytokines and their roles
in pancreatic islet beta-cell destruction and insulin-dependent
diabetes mellitus. Biochem Pharmacol 1998;55:11391149.
[41] Adorini L, Trembleau S. Immune deviation towards Th2
inhibits Th-1-mediated autoimmune diabetes. Biochem Soc
Trans 1997;25:625629.
[42] Liblau RS, Singer SM, McDevitt HO. Th1 and Th2
CD4+Tcells in the pathogenesis of organ-specific auto-
immune diseases. Immunol Today 1995;16:34 38.
[43] Rapoport MJ, Jaramillo A, Zipris D, et al. Interleukin 4
reverses T-cell proliferative unresponsiveness and prevents the
onset of diabetes in nonobese diabetic mice. J Exp Med
1993;178:8799.
[44] Cameron MJ, Arreaza GA, Zucker P, et al. IL-4 prevents
insulitis and insulin-dependent diabetes mellitus in nonobese
diabetic mice by potentiation of regulatory T helper-2 cell
function. J Immunol 1997;159:4686 4692.
[45] Tominaga Y, Nagata M, Yasuda H, et al. Administration of
IL-4 prevents autoimmune diabetes but enhances pancreatic
insulitis in NOD mice. Clin Immunol Immunopathol
1998;86:209218.
[46] Mueller R, Krahl T, Sarvetnick N. Pancreatic expression of
interleukin-4 abrogates insulitis and autoimmune diabetes in
nonobese diabetic (NOD) mice. J Exp Med
1996;184:1093 1099.
[47] Rabinovitch A, Suarez-Pinzon WL, Sorensen O, Bleackley
RC, Power RF, Rajotte RV. Combined therapy with
interleukin-4 and interleukin-10 inhibits autoimmune diabetes
recurrence in syngeneic islet-transplanted nonobese diabetic
mice. Analysis of cytokine mRNA expression in the graft.
Transplantation 1995;60:368374.
[48] Kolb H. Benign versus destructive insulitis. Diabetes Metab
Rev 1997;13:139146.
[49] Kaufman DL, Clare-Salzler M, Tian J, et al. Spontaneous loss
of T-cell tolerance to glutamic acid decarboxylase in murine
insulin-dependent diabetes. Nature 1993;366:69 72.
[50] Tian J, Chau C, Kaufman DL. Insulin selectively primes Th2
responses and induces regulatory tolerance to insulin in
pre-diabetic mice. Diabetologia 1998;41:237 240.
Cannabidiol lowers incidence of diabetes 151
... <20 µM [79][80][81] IL-4 and IL-10 Increases levels Anti-inflammatory cytokines. 5 mg/Kg (in vivo) [82,83] ROS Inhibitor CBD inhibits a mechanism related to NADPH oxidase-mediated ROS production and NF-κB-dependent signaling events <10 µM [84,85] iNOS and COX2 Inhibition of expression ...
Article
Full-text available
The phytocannabinoid cannabidiol (CBD) is receiving increasing attention due to its pharmacological properties. Although CBD is extracted from Cannabis sativa, it lacks the psychoactive effects of Δ9-tetrahydrocannabinol (THC) and has become an attractive compound for pharmacological uses due to its anti-inflammatory, antioxidant, anticonvulsant, and anxiolytic potential. The molecular mechanisms involved in CBD’s biological effects are not limited to its interaction with classical cannabinoid receptors, exerting anti-inflammatory or pain-relief effects. Several pieces of evidence demonstrate that CBD interacts with other receptors and cellular signaling cascades, which further support CBD’s therapeutic potential beyond pain management. In this review, we take a closer look at the molecular mechanisms of CBD and its potential therapeutic application in the context of cancer, neurodegeneration, and autoimmune diseases.
... And oral routes were found to result in similar concentrations in the rat (both in the plasma and brain) (Deiana et al., 2012). In diabetic rodent models, the duration of CBD treatment varied between 1 and 11 weeks(s), but the most frequent length was 4 weeks (El-Remessy et al., 2006;Weiss et al., 2006Weiss et al., , 2008Rajesh et al., 2010;Santiago et al., 2019). Taking all together, we decided to use 60 mg/kg/day CBD, per os, for 4 weeks, considering both the dose and duration to be sufficient to act in a well-detectable but safe (Dziwenka et al., 2020) manner. ...
Article
Full-text available
Cannabidiol (CBD), the most extensively studied non-intoxicating phytocannabinoid, has been attracting a lot of interest worldwide owing to its numerous beneficial effects. The aim of this study was to explore the effect that CBD exerts on the adenosinergic system of paced left atria isolated from obese type Zucker Diabetic Fatty (ZDF) rats, maintained on diabetogenic rat chow, received 60 mg/kg/day CBD or vehicle via gavage for 4 weeks. We found that N6-cyclopentyladenosine (CPA), a relatively stable and poorly transported A1 adenosine receptor agonist, elicited a significantly weaker response in the CBD-treated group than in the vehicle-treated one. In contrast, adenosine, a quickly metabolized and transported adenosine receptor agonist, evoked a significantly stronger response in the CBD-treated group than in the vehicle-treated counterpart (excepting its highest concentrations). These results can be explained only with the adenosine transport inhibitory property of CBD (and not with its adenosine receptor agonist activity). If all the effects of CBD are attributed to the interstitial adenosine accumulation caused by CBD in the myocardium, then a significantly increased adenosinergic activation can be assumed during the long-term oral CBD treatment, suggesting a considerably enhanced adenosinergic protection in the heart. Considering that our results may have been influenced by A1 adenosine receptor downregulation due to the chronic interstitial adenosine accumulation, an adenosinergic activation smaller than it seemed cannot be excluded, but it was above the CBD-naïve level in every case. Additionally, this is the first study offering functional evidence about the adenosine transport inhibitory action of CBD in the myocardium.
... Cannabidiol (CBD) is a nonintoxicating constituent of the cannabis plant and gained a lot of attention in clinical populations because of its analgesic, 1 neuroprotective, 2,3 anti-inflammatory, [4][5][6][7][8] and antioxidative 9,10 effects with a safe adverse event profile. 11,12 This has contributed to the recent approval of CBD to treat epilepsy and tuberous sclerosis. ...
Article
Background: Cannabidiol (CBD), a nonintoxicating constituent of the cannabis plant, recently gained a lot of interest among athletes, since it is no longer considered as a prohibited substance by the World Anti-Doping Agency. The increasing prevalence of CBD use among athletes is driven by a perceived improvement in muscle recovery and a reduction in pain. However, compelling evidence from intervention studies is lacking and the precise mechanisms through which CBD may improve muscle recovery remain unknown. This highlights the need for more scientific studies and an evidence-based background. In the current review, the state-of-the-art knowledge on the effects of CBD on skeletal muscle tissue is summarized with special emphasis on the underlying mechanisms and molecular targets. More specifically, the large variety of receptor families that are believed to be involved in CBD's physiological effects are discussed. Furthermore, in vivo and in vitro studies that investigated the actual effects of CBD on skeletal muscle metabolism, inflammation, tissue regeneration, and anabolism are summarized, together with the functional effects of CBD supplementation on muscle recovery in human intervention trials. Overall, CBD was effective to increase the expression of metabolic regulators in muscle of obese mice (e.g., Akt, glycogen synthase kinase-3). CBD treatment in rodents reduced muscle inflammation following eccentric exercise (i.e., nuclear factor kappa B [NF-κB]), in a model of muscle dystrophy (e.g., interleukin-6, tumor necrosis factor alpha) and of obesity (e.g., COX-2, NF-κB). In addition, CBD did not affect in vitro or in vivo muscle anabolism, but improved satellite cell differentiation in dystrophic muscle. In humans, there are some indications that CBD supplementation improved muscle recovery (e.g., creatine kinase) and performance (e.g., squat performance). However, CBD doses were highly variable (between 16.7 and 150 mg) and there are some methodological concerns that should be considered. Conclusion: CBD has the prospective to become an adequate supplement that may improve muscle recovery. However, this research domain is still in its infancy and future studies addressing the molecular and functional effects of CBD in response to exercise are required to further elucidate the ergogenic potential of CBD.
... Comparison of FBS values across groups strongly suggests that oral CBD on its own can stabilize blood glucose. Most studies on the hypoglycaemic effects of CBD have reported similar findings, though experimental designs and doses varied [77][78][79] . Mechanisms of action may involve stimulation of insulin production by the beta-pancreatic cells and protective and regenerative effects on the beta cells 80,81 . ...
Article
Full-text available
Atypical antipsychotics, despite their rapid dissociation from dopamine receptors and reduced tendency to induce oxidative stress, have been associated with difficult-to-manage movement disorders, including tardive dyskinesia (TD). The study set out to investigate the effects of cannabidiol (CBD), a potent antioxidant, on risperidone-induced behavioural and motor disturbances; namely vacuous chewing movements (VCM), and oxidative stress markers (e.g. superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), malondialdehyde (MDA), Nitric oxide (NO), and DPPH (2,2-diphenyl-1-picrylhydrazyl)). Oral risperidone (10 mg/kg) or oral CBD (5 mg/kg) were administered to six experimental groups. While risperidone alone was administered for 28 days, CBD concomitantly or in sequential order with risperidone, was administered for 28 days; and CBD alone was administered for 21 days. Behavioural, motor, and specific biochemical parameters, which included VCM, muscle tone, fasting blood sugar (FBS), and oxidative stress markers were assessed at different time points after the last dose of medication. Oral CBD (5 mg/kg) significantly reduced risperidone-induced elevated FBS when given after the administration of risperidone. Oral CBD also had effects on VCM when administered before risperidone and similarly, attenuated risperidone-induced increased muscle tone. It was also established that concomitant or sequential administration of CBD and risperidone did not have any adverse effects on cognition or locomotion. Both CBD and risperidone increased the activity of antioxidant enzymes and decreased the activity of pro-oxidant enzymes. This study suggests CBD could mitigate metabolic dysregulation and extrapyramidal side effects associated with risperidone without producing cognitive impairments.
... It acts as antagonist for CB1 and CB2 receptors, causing anxiolytic, anti-sedative, and anti-psychotic effects [14]. In addition, it has the potential to cause many other effects such as anti-inflammatory effects [15] and prevents cancer cell growth [16] and the neuroprotection of neuro-degenerative diseases such as postischemia and Parkinson's syndrome [17]. In addition, the use of cannabis has some adverse effects, which preclude its widespread use as a medicinal agent. ...
Article
Full-text available
The development of new antibiotics is urgently needed to combat the threat of bacterial resistance. New classes of compounds that have novel properties are urgently needed for the development of effective antimicrobial agents. The extract of Cannabis sativa L. has been used to treat multiple ailments since ancient times. Its bioactivity is largely attributed to the cannabinoids found in its plant. Researchers are currently searching for new anti-infective agents that can treat various infections. Although its phytocannabinoid ingredients have a wide range of medical benefits beyond the treatment of infections, they are primarily associated to psychotropic effects. Different cannabinoids have been demonstrated to be helpful against harmful bacteria, including Gram-positive bacteria. Moreover, combination therapy involving the use of different antibiotics has shown synergism and broad-spectrum activity. The purpose of this review is to gather current data on the actions of Cannabis sativa (C. sativa) extracts and its primary constituents such as terpenes and cannabinoids towards pathogens in order to determine their antimicrobial properties and cytotoxic effects together with current challenges and future perspectives in biomedical application.
Article
Full-text available
The use of marijuana in medication has been criminalized in Indonesia since 1997. Twenty years after, Fidelis Arie Sudewarto violated the rule by administering marijuana as an alternative means of medication for his spouse, Yeni Riawati. Various public opinions emerged, indicating a shift of paradigm on the use of marijuana for medication, and this affects the fulfillment of "unlawful nature" which is expressly stated as one of the elements in the formulation of offense as stipulated in Article 116 paragraph (2) Law Number 35 of 2009, especially in assessing the material unlawfulness nature (materieele wederrechtelijkheid) of the act. This study discusses the case of Fidelis from the perspective of criminal law, namely how the fulfillment of the element of unlawful nature and whether there is a basis which then abolish the unlawful nature in the acts committed by Fidelis Arie Sudewarto.
Article
Background: With changes in state laws, cannabis is now more accessible in many parts of the United States. The drug has previously been associated with a higher incidence of acute ischemic stroke (AIS). Our study analyzed the effect of cannabis use on AIS outcomes. Methods: The 2019 National Inpatient Sample from Healthcare Cost and Utilization Project and Agency for Healthcare Research and Quality was used to evaluate the AIS patients ages 25 and more. Cannabis users were compared with nonusers. Their mortality risks were also assessed. Results: Five hundred thirty-nine thousand eight hundred eighty patients with AIS were in our study and 8850 used cannabis. Among the cannabis users with AIS, 69.8% were males, 50.6% were white, 61.8% were aged 46 to 65, 32.1% used Medicaid, 86.1% were nonobese, 72.0% were nondiabetics, 81.7% had hypertension, 80.7% did not have a history of alcohol abuse, and 77.5% were smokers (nicotine). Only 12.8% of cannabis users reported any cardiac dysrhythmia, whereas 5.4% required mechanical thrombectomy, and 8.0% received intravenous tissue plasminogen activator (tPA).After adjusting the variables, cannabis users covered by Medicare, with diabetes, a history of alcohol abuse, cardiac dysrhythmias, and who underwent mechanical thrombectomy had a higher risk of dying. The overall adjusted odds ratio of dying among cannabis users with AIS was 0.603 (95% CI, 0.504-0.722, P<0.01). Conclusions: Although the overall mortality risk seems to be lower among cannabis users, those with certain risk factors still present higher mortality risks. Patients with cannabis use should therefore be monitored closely for possible complications and mortality after AIS.
Article
The prevalence of obesity and insulin‐resistance is on the rise, globally. Cannabis have been shown to have anti‐diabetic/obesity properties, however, the effect mediated at various fat depots remains to be clarified. The aim of this study was to (1) investigate the anti‐diabetic property of an oral cannabis administration in an obese and streptozotocin‐induced diabetic rat model and (2) to determine and compare the effect mediated at the peritoneal and intramuscular fat level. Cannabis concentration of 1.25 mg/kg body weight (relative to THC content) was effective in reversing insulin‐resistance in the rat model, unlike the other higher cannabinoid concentrations. At the peritoneal fat level, gene expression of fat beigeing markers, namely Cidea and UCP1, were significantly increased compared to the untreated control. At the intramuscular fat level, on the other hand, CE1.25 treatment did not promote fat beigeing but instead significantly increased mitochondrial activity, relative to the untreated control. Therefore, these findings indicate that the mechanism of action of oral cannabis administration, where glucose and lipid homeostasis is restored, is not only dependent on the dosage but also on the type of fat depot investigated.
Article
The number of people diagnosed with diabetes mellitus and its complications is markedly increasing worldwide, leading to a worldwide epidemic across all age groups, from children to older adults. Diabetes is associated with premature aging. In recent years, it has been found that peripheral overactivation of the endocannabinoid system (ECS), and in particular cannabinoid receptor 1 (CB1R) signaling, plays a crucial role in the progression of insulin resistance, diabetes (especially type 2), and its aging-related comorbidities such as atherosclerosis, nephropathy, neuropathy, and retinopathy. Therefore, it is suggested that peripheral blockade of CB1R may ameliorate diabetes and diabetes-related comorbidities. The use of synthetic CB1R antagonists such as rimonabant has been prohibited because of their psychiatric side effects. In contrast, phytocannabinoids such as cannabidiol (CBD) and tetrahydrocannabivarin (THCV), produced by cannabis, exhibit antagonistic activity on CB1R signaling and do not show any adverse side effects such as psychoactive effects, depression, or anxiety, thereby serving as potential candidates for the treatment of diabetes and its complications. In addition to these phytocannabinoids, cannabis also produces a substantial number of other phytocannabinoids, terpenes, and flavonoids with therapeutic potential against insulin resistance, diabetes, and its complications. In this review, the pathogenesis of diabetes, its complications, and the potential to use cannabinoids, terpenes, and flavonoids for its treatment are discussed.
Book
Full-text available
Kenevirin tıpta yeri
Article
Full-text available
To further characterize neuronal cannabinoid receptors, we compared the ability of known and novel cannabinoid analogs to compete for receptor sites labeled with either [3H]SR141716A or [3H]CP-55,940. These efforts were also directed toward extending the structure-activity relationships for cannabinoid agonists and antagonists. A series of alternatively halogenated analogs of SR141716A were synthesized and tested in rat brain membrane binding assays along with the classical cannabinoids, Delta9-tetrahydrocannabinol, cannabinol, cannabidiol, the nonclassical cannabinoid CP-55,940, the aminoalkylindole WIN55212-2 and the endogenous fatty acid ethanolamide, anandamide. Saturation binding isotherms were performed with both radioligands, as were displacement studies, allowing an accurate comparison to be made between the binding of these various compounds. Competition studies demonstrated that all of the compounds were able to displace the binding of [3H]CP-55,940 with rank order potencies that agreed with previous studies. However, the rank order potencies of these compounds in competition studies with [3H]SR141716A differed significantly from those determined with [3H]CP-55,940. These results suggest that CP-55,940, WIN55212-2 and other agonists interact with cannabinoid binding sites within the brain which are distinguishable from the population of binding sites for SR141716A, its analogs and cannabidiol. Structural modification of SR141716A significantly altered the affinity of the compound and its relative ability to displace either [3H]CP-55,940 or [3H]SR141716A preferentially within the rat brain receptor membrane preparation.
Article
The in vitro production of interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha) by monocytes was examined in patients with insulin-dependent diabetes mellitus (IDDM), in those with noninsulin-dependent diabetes mellitus (NIDDM), and in healthy volunteers. The production of IL-1 and IL-6 by monocytes was significantly lower in IDDM patients than in NIDDM patients and normal subjects whereas the TNF-alpha production by monocytes did not differ between IDDM patients and normal subjects. On the other hand, the TNF-alpha production was significantly higher in NIDDM patients than in IDDM patients and normal subjects. There was a significant correlation between IL-1 and IL-6 concentrations in culture supernatants of monocytes for IDDM patients but not for NIDDM patients and normal subjects. Neither glucose nor insulin showed any stimulatory effect on in vitro production of these monokines. In the serial observation lasting 3-18 months, the monocyte production of IL-1 was found ...
Article
The Diabetes Control and Complications Trial has demonstrated that intensive diabetes treatment delays the onset and slows the progression of diabetic complications in subjects with insulin-dependent diabetes mellitus from 13 to 39 years of age. We examined whether the effects of such treatment also occurred in the subset of young diabetic subjects (13 to 17 years of age at entry) in the Diabetes Control and Complications Trial. One hundred twenty-five adolescent subjects with insulin-dependent diabetes mellitus but with no retinopathy at baseline (primary prevention cohort) and 70 adolescent subjects with mild retinopathy (secondary intervention cohort) were randomly assigned to receive either (1) intensive therapy with an external insulin pump or at least three daily insulin injections, together with frequent daily blood-glucose monitoring, or (2) conventional therapy with one or two daily insulin injections and once-daily monitoring. Subjects were followed for a mean of 7.4 years (4 to 9 years). In the primary prevention cohort, intensive therapy decreased the risk of having retinopathy by 53% (95% confidence interval: 1% to 78%; p = 0.048) in comparison with conventional therapy. In the secondary intervention cohort, intensive therapy decreased the risk of retinopathy progression by 70% (95% confidence interval: 25% to 88%; p = 0.010) and the occurrence of microalbuminuria by 55% (95% confidence interval: 3% to 79%; p = 0.042). Motor and sensory nerve conduction velocities were faster in intensively treated subjects. The major adverse event with intensive therapy was a nearly threefold increase of severe hypoglycemia. We conclude that intensive therapy effectively delays the onset and slows the progression of diabetic retinopathy and nephropathy when initiated in adolescent subjects; the benefits outweigh the increased risk of hypoglycemia that accompanies such treatment. (J PEDIATR 1994;125:177-88)
Article
BACKGROUND Long-term microvascular and neurologic complications cause major morbidity and mortality in patients with insulin-dependent diabetes mellitus (IDDM). We examined whether intensive treatment with the goal of maintaining blood glucose concentrations close to the normal range could decrease the frequency and severity of these complications. METHODS A total of 1441 patients with IDDM -- 726 with no retinopathy at base line (the primary-prevention cohort) and 715 with mild retinopathy (the secondary-intervention cohort) were randomly assigned to intensive therapy administered either with an external insulin pump or by three or more daily insulin injections and guided by frequent blood glucose monitoring or to conventional therapy with one or two daily insulin injections. The patients were followed for a mean of 6.5 years, and the appearance and progression of retinopathy and other complications were assessed regularly. RESULTS In the primary-prevention cohort, intensive therapy reduced the adjusted mean risk for the development of retinopathy by 76 percent (95 percent confidence interval, 62 to 85 percent), as compared with conventional therapy. In the secondary-intervention cohort, intensive therapy slowed the progression of retinopathy by 54 percent (95 percent confidence interval, 39 to 66 percent) and reduced the development of proliferative or severe nonproliferative retinopathy by 47 percent (95 percent confidence interval, 14 to 67 percent). In the two cohorts combined, intensive therapy reduced the occurrence of microalbuminuria (urinary albumin excretion of ≥ 40 mg per 24 hours) by 39 percent (95 percent confidence interval, 21 to 52 percent), that of albuminuria (urinary albumin excretion of ≥ 300 mg per 24 hours) by 54 percent (95 percent confidence interval, 19 to 74 percent), and that of clinical neuropathy by 60 percent (95 percent confidence interval, 38 to 74 percent). The chief adverse event associated with intensive therapy was a two-to-threefold increase in severe hypoglycemia. CONCLUSIONS Intensive therapy effectively delays the onset and slows the progression of diabetic retinopathy, nephropathy, and neuropathy in patients with IDDM.
Article
The non-obese diabetic (NOD) mouse represents a relevant animal model of autoimmunity for insulin-dependent diabetes mellitus. The pathogenic role of tumor necrosis factor (TNF) in insulitis and β cell destruction observed in these mice remains controversial, since injections of TNF or of anti-TNF antibodies have been reported to exert protection or acceleration of diabetes, depending on the timing of administration. In this study, we demonstrate that, in contrast to the non-transgenic littermates, NOD mice with permanent neutralization of TNF by high blood levels of soluble TNF receptor p55-human FcIgG3-fusion molecules resulting from the expression of a transgene are protected from spontaneous diabetes. They are also protected from accelerated forms of disease caused by transfer of NOD spleen cells or cyclophosphamide injections. This protection is associated with a marked decrease in the severity and incidence of insulitis and in the expression of the adhesion molecules MAdCAM-1 and ICAM-1 on the venules of pancreatic islets. These data suggest a central role for TNF-α in the mediation of insulitis and of the subsequent destruction of insulin-secreting β-cells observed in NOD mice. They may be relevant to cell-mediated autoimmune diseases in general, in which treatment with soluble TNF receptors might be beneficial.
Article
The present study demonstrated that the administration of recombinant interleukin-4 (rIL-4) prevented overt diabetes in nonobese diabetic (NOD) mice whose T cells produced relatively low amounts of IL-4. However, massive insulitis was observed in rIL-4-treated NOD mice. The flow cytometric analysis of islet-infiltrating T cells revealed that the number of CD45RBlowCD4+T cells was significantly increased byin vivoadministration of rIL-4. By measuring the cytokine production of splenic T cells after stimulation, it was shown that CD45RBlowCD4+T cells predominantly produced IL-4 and IL-10 but produced less IL-2 and interferon-γ (IFN-γ). A semiquantitative reverse-transcriptase polymerase chain reaction assay revealed a higher expression of IL-4 and IL-10 mRNA and an apparent decrease in IFN-γ mRNA in the islets of NOD mice which were administered rIL-4. These results suggested that autoreactive CD45RBlowCD4+T helper 2 (Th2)-like cells which developed following rIL-4 administration were predominant in the infiltrate of the islets, and overt diabetes was prevented. On the other hand, when splenocytes from rIL-4-treated NOD mice were transferred to irradiated NOD recipients, along with splenocytes from diabetic NOD mice, all of the recipient mice became diabetic within 8 weeks after transfer. Considered together, a supplement of rIL-4 administered to NOD mice may protect against autoimmune diabetes by facilitating the development of Th2-like autoreactive T cells in the islets.
Article
Marijuana cannabinoids are both psychoactive and immunoactive. Here, we will review evidence that cannabinoids modulate immunity and that cannabinoid receptors and endogenous ligands are expressed in immune tissues. Clues will also be presented concerning the role of the cannabinoid system in immune regulation and the possible molecular mechanisms involved.
Article
Little is known about the immunological impact of insulin administration other than it can boost insulin autoantibody levels. In particular, while the subcutaneous administration of a soluble foreign antigen (without adjuvant) is generally only weakly immunogenic in a naive animal, it is unknown what effect the subcutaneous administration of a soluble self-antigen has in animals with established autoimmune responses to the antigen. Addressing these questions in pre-diabetic nonobese diabetic (NOD) mice, we examined the effects of administering insulin, as well as the metabolically inactive B-chain of insulin, on insulin-specific cellular and humoral immune responses. We show that pre-diabetic NOD mice have a spontaneous Th1-biased response against insulin. Administering insulin, or the insulin B-chain, rather than boosting the established Th1 response, primed Th2 cellular and humoral immunity to insulin, shifting the predominant insulin response toward a Th2 phenotype. Despite the presence of a Th1 response against insulin, insulin treated mice failed to mount proliferative T-cell responses following immunization and challenge with insulin, demonstrating that the treatment induced an active form of tolerance to this autoantigen. Thus, the subcutaneous administration of a soluble antigen can engage Th2 responses and induce self-tolerance, even after the establishment of autoreactive Th-1 responses. Such immune deviation and induced regulatory tolerance may contribute to the protective effects of prophylactic insulin therapy, as well as the establishment of a "honeymoon" phase in new-onset insulin-dependent diabetic patients.