Current Pharmaceutical Design, 2008, 14, 2289-2298 2289

Expression and Function of the Endocannabinoid System in Glial Cells

Paola Massi1, Marta Valenti2, Daniele Bolognini2 and Daniela Parolaro2,*

1
Department of Pharmacology, Chemotherapy and Toxicology, University of Milan, Via Vanvitelli 32, 20129 Milan,
Italy and 2Department of Structural and Functional Biology, Center of Neuroscience, University of Insubria, Via A. da
Giussano 10, 21052 Busto Arsizio (Varese), Italy

Abstract: In the last few years the role and significance of the glia in CNS function and pathology have been drastically
reassessed. Glial cells physiology appears very different in healthy versus pathological brain and the recent identification
of cannabinoid receptors and their endogenous ligands in glia has triggered a number of studies exploring the role of
(endo)cannabinoid system in glia functionality and disease. (Endo)cannabinoids exert their effects in these cells directly
affecting some important peculiar functions of the glia and actively promoting biochemical signals ending in a pro-
survival fate for these cells. By contrast, (endo)cannabinoids induce a selective death in glia-derived tumor cells. Of spe-
cial physiological and therapeutic relevance is the reported ability of glial cells during neuropathological conditions to re-
lease an increased amount of endocannabinoids and to overexpress cannabinoid receptors. This evidence has suggested
that the endocannabinoids production by glial cells may constitute an endogenous defense mechanism preventing the
propagation of neuroinflammation and cell damage. The present paper will review the evidence supporting the regulatory
role of (endo)cannabinoids in glia function, holding in consideration their therapeutic potential as neuroprotective and/or
anticancer agents.
Key Words: Glia, microglia, astrocytes, oligodendrocytes, neuroprotection, cell survival, glioma.

INTRODUCTION
The endocannabinoid system (ECs) is made up of en-
dogenous ligands (endocannabinoids) derived from arachi-
donic acid, and their specific receptors that are expressed and
present in all mammalian tissues. The highly conserved na-
ture of the ECs and its wide distribution, particularly in the
central nervous system (CNS), suggest it is an important
biological regulatory system. So far, the ECs has been impli-
cated in a variety of brain functions, such as the control of
pain, memory, movement, thermoregulation, and addictive
behavior. It appears to have a fundamental role as a neuro-
protective system through its action on synaptic plasticity
and also through the control of glial cell responses.
In the last few years the role and significance of the glia
in CNS function and pathology have been drastically reas-
sessed, so some published results about the relationship be-
tween the ECs and its effects on glial response add very im-
portant knowledge of the role of the ECs in CNS functional-
ity and disease. This is a growing, promising field of re-
search.
THE CELLS OF THE GLIA
Scientific inquiry into the biology of glia suffered an
eclipse that lasted 50 years, but recognition towards the end
of the 20th century has opened up fresh investigations into
its place in the CNS, indicating a pivotal role in the physiol-
ogy as well as in many, if not all, neuroinflammatory/ neu-
rodegenerative diseases of the brain.
*Address correspondence to this author at the Department of Structural and
Functional Biology, Center of Neuroscience, University of Insubria, Via A.
da Giussano 10, 21052 Busto Arsizio (VA), Italy;
E-mail: daniela.parolaro@uninsubria.it
There are two groups of glial cells in the CNS, the mi-
croglia and the macroglia, including astrocytes, oligodendro-
cytes, and ependymal cells. The most important cells in the
maintenance of homeostasis and therefore in some brain dis-
eases are microglia, astrocytes and oligodendrocytes. Micro-
glia are considered the resident immune cells of the central
nervous system, and have been called the “resident and pro-
fessional macrophages of the nervous system” on account of
their phenotype and reactivity in response to injury and in-
flammation [1-3].
Under normal conditions in the adult microglia - some-
times called “resting” microglia- are distributed throughout
the nervous system. They have a small cell body with fine,
highly ramified morphology and minimal expression of sur-
face antigens (Fig. 1).
In pathological conditions, microglia can become pro-
gressively more activated, retracting cellular processes and
expressing distinct profile markers, thus exhibiting the full
range of immunological surveillance. Microglial activation
involves increased cell recruitment, primarily through mito-
sis of resident microglia but also from bone-marrow-derived
precursors that infiltrate the CNS. Activation of these cells
causes their transformation into phagocytic cells able to pre-
sent antigen to circulating T cells, and to produce a variety of
inflammatory cytokines, reactive oxygen species (ROS) and
nitric oxide (NO). Microglial cells can also process and pre-
sent epitopes in association with Major Histocompatibility
Complex (MHC) class II molecules to CD4 + T cells within
the CNS (Fig. 1). These responses represent important de-
fense functions against invading neurotropic pathogens and
have been implicated in brain damage in infectious and neu-
roinflammatory/ neurodegenerative diseases such as multiple

1381-6128/08 $55.00+.00 © 2008 Bentham Science Publishers Ltd.

2290 Current Pharmaceutical Design, 2008, Vol. 14, No. 23 Massi et al.

Fig. (1). Schematic representation of the role of ECs in glia under pathological conditions in the brain.
When an injury or inflammatory events occur, there is from neurons and especially from glial cells, a sustained production and release of
endocannabinoids (EC). The EC as well as exogenous cannabinoids can bind cannabinoid receptors (CBr) present on glia. This event lead to
a decreased release of NO, diminished production of reactive oxygen species (ROS), inhibition of the release of pro-inflammatory cytokines
from microglia and astrocytes, and increasing remyelination by oligodendrocytes. In addition, EC modulate the glial cells survival activating
some defined intracellular pathways (see text for more details). All together, these events mediate anti-inflammatory and neuroprotective
effects.

sclerosis (MS), Alzheimer's disease (AD), Parkinson's dis-
ease, and amyotrophic lateral sclerosis (ALS) (see the spe-
cific chapters in the present review). In this activated state,
microglia can contribute to or exacerbate neuronal damage
although, under certain conditions, they may also have a
neuroprotective and neurotrophic function.
Astrocytes are the major glial cell within the CNS and
have a number of important physiological properties related
to CNS homeostasis. They affect neuronal function by the
release of neurotransmitters and neurotrophic factors, guide
neuronal development, contribute to the metabolism of
neurotransmitters, regulate extracellular pH and K+ levels [4-
6] and play a critical role in the regulation of brain metabolic
responses [7]. It has been proposed that neurons and astro-
cytes are an integral unit with distinct roles in different fun-
damental events in brain function [6]. Some data suggest that
soluble factors secreted by astrocytes are involved in main-
tenance of the blood/brain barrier, probably by inducing the
formation of tight junctions between endothelial cells [8].
Another important, recently discovered aspect of the biology
of astrocytes is their ability to behave as immunocompetent
cells within the brain. Although it is still debated, there is
evidence of the astrocytes’ ability to express MHC class II
antigens (Fig. 1) and co-stimulatory molecules (B7 and
CD40) that are essential for antigen presentation and T-cell
activation and there is no doubt about their ability to produce
Expression and Function of the Endocannabinoid System in Glial Cells
a wide variety of cytokines and chemokines. The expression
of these molecules in Parkinson’s disease, MS, AD and brain
injury/trauma is well documented. Thus, as a potent source
of immunologically important chemokines and cytokines,
astrocytes are thought to play a major role in the type and
extent of CNS diseases.
Oligodendrocytes, or oligodendroglia, are a variety of
macroglia. Besides the Schwann cells in the peripheral nerv-
ous system, oligodendrocytes are the fundamental cells that
coat CNS axons with their membrane, called myelin. This
myelin sheath provides insulation to the axon so electrical
signals can propagate more efficiently (Fig. 1). As part of the
nervous system they are closely related to nerve cells and,
like all other glial cells, the oligodendrocytes have a support-
ing role towards neurons.
Diseases that result in injury to the oligodendroglial cells
include demyelinating diseases such as cerebral palsy, spinal
cord injury, stroke and possibly MS, where oligodendrocytes
are thought to be damaged by excessive release of the neuro-
transmitter glutamate.
THE ENDOCANNABINOID SYSTEM IN GLIA
Glia possess all the biochemical machinery required for
endocannabinoid synthesis, transport, and degradation.
These findings, together with evidence that the ECs is in-
volved in the pathophysiology of CNS inflammatory and
degenerative disorders, have cast fresh light on the place of
this system in brain diseases.
CB1 Receptors
Although the presence of CB1 receptors in glial cells has
been debated in the past, they have been recently described.
With an in situ approach Rodriguez et al. [9] proved that
glial cells express CB1 receptors; perivascular astrocytes
express these receptors and rat astrocytes in the nucleus ac-
cumbens express them too, along the plasmalemma of their
perivascular end-feet. CB1 receptors are also present in the
astrocytes of the amygdala, cingulate cortex, median fore-
brain bundle, and lamina I and II of the spinal cord of Wistar
rats [10,11], in the human astrocytoma cells [12], in C6
glioma cells [13], in mouse spinal neurons [14], in rat oli-
godendrocytes [15] and recently, Navarrete and Araque [16]
found them in hippocampal astrocytes derived from mice.
However, besides the above cited reports, other groups did
not find them in cultured astrocytes [17,18]. These conflict-
ing results may reflect differences in cell culture conditions,
animal species, age and differentiation state, and also the
CNS structures used to prepare the astrocyte cultures (rat
hippocampus [12], rat cortex [13], mouse spinal cord [14],
rat forebrain [15], C57BL/6 cortex [17], cortex and striatum
of rat and mouse [18]). In any case, CB1 receptors are ex-
pressed in astrocytes at a lower level than in neurons. While
in neurons CB1 activation leads to inhibition of transmitter
release [19], in primary astrocytes derived from cortices of
rat these receptors are involved in the brain’s energy supply,
increasing the rate of glucose oxidation and ketogenesis and
regulating cell survival [20,21].
The CB1 receptor is also detected in microglial cells, the
immunocompetent CNS cells that are specifically subject to
Current Pharmaceutical Design, 2008, Vol. 14, No. 23 2291
major phenotypic and genotypic transformation during their
activation in response to stress and injury. Although it is not
clear whether CB1 expression is constant throughout the
different states of activation, Maresz et al. [22] reported no
change in the expression of CB1 receptor mRNA in micro-
glia during the progression of experimental autoimmune
encephalomyelitis (EAE). The functional significance of
these receptors in microglia is still not certain, because they
are expressed at low levels and are located predominantly in
the intracellular compartments [23]. Some groups [24] have
proposed a role of CB1 receptors in NO production, a char-
acteristic of the microglial response in neuroinflammation.
These receptors have also been detected in oligodendrocytes
in vivo and in culture [15].
CB1 receptor immunoreactivity in white matter has been
associated mostly with postnatal and adult brain oligoden-
drocytes and, when cultured, these cells have lower CB1
protein and mRNA levels than neurons but higher than as-
trocytes and microglia [15]. These findings have suggested
the cannabinoids may play an important role in remyelinat-
ing processes after an inflammatory insult.
CB2 Receptors
For over a decade, since the cloning of the CB2 receptor,
several laboratories have still not detected CB2 cannabinoid
receptors in healthy brain, in notable contrast to the well-
known abundant distribution of CB1 receptors in the CNS
[25-28]. However, even if there are still some controversies
on the specificity of CB2 receptor antibodies staining, a large
number of recent reports have described the presence of CB2
receptors in the brain using CB2 receptor antibodies [29-31],
CB2 gene expression/level detection [32] and pharmacologi-
cal approaches [16]. CB2 positive neurons have been re-
ported in the vagus nerve in the brainstem [33], in other
brain structures such as cerebellum, olfactory tubercle, cere-
bral cortex, striatum, thalamic nuclei, hippocampus, amyg-
dala, substantia nigra [34] and in primary cultured neurons
[35]. In contrast, other reports describe CB2 receptor as es-
sentially localized in situ in cerebral microvasculature [36]
and in cultured human cerebral microvessel endothelia [37].
However, there is increasing and consistent evidence that
CB2 in the brain is substantially restricted to cells of the
immune lineage [38].
Glial cells, which can be considered parenchymal and
immunocompetent brain tissue, can express CB2 receptors
[34]. Although almost absent in the healthy CNS, CB2 re-
ceptors have been found in perivascular microglia in human
cerebellum [39]. Their presence in this district might be re-
lated to their involvement in different immunomodulatory
processes and a role in regulating the blood/brain barrier
function has been suggested. Carlisle et al. [28] reported that
also rat neonatal microglial cells express CB2. These recep-
tors seem to play an important role in microglia migration
[23] and proliferation [40].
Activated microglia express CB2 receptors in response to
different conditions associated with local inflammatory
events and injury. Benito et al. [29] reported the expression
of CB2 receptors in specific subtypes of glial cells in AD
tissue samples. In ß-amyloid (ßA) plaque-rich regions, acti-
vated microglia were highly immunopositive for CB2 recep-
2292 Current Pharmaceutical Design, 2008, Vol. 14, No. 23
tors. CB2 but not CB1 receptors are overexpressed in the
brains of
A–treated rats and of AD patients [29, 30]. This
pattern of cell type expression is seen in other conditions
involving marked neuroinflammation too. In simian immu-
nodeficiency virus-induced encephalitis (SIVE), microglial
expression of the CB2 receptor is increased [41] as in pa-
tients with MS [42]. CB2 receptors are also induced in ßA
plaque–associated microglia in Down’s syndrome (DS), a
pathology sometimes considered as a human model of Alz-
heimer-like ß-amyloid deposition, while the signal is almost
undetectable in control samples and young DS patients [31].
These results further confirm that CB2 receptors are ex-
pressed at very low levels in healthy brain and are up-
regulated in inflamed and injured brain only in glial cells, as
previously suggested [43,44].
There is only limited data on the presence of cannabinoid
receptors in oligodendrocytes which undergo degeneration in
MS and EAE. Previous reports [45] indicate low expression
of the cannabinoid receptors, at least in healthy conditions.
However, Molina-Holgado’s group [15] reported that oli-
godendrocyte progenitors and differentiated oligodendro-
cytes express CB2 receptors in culture. This suggests that
CB2 receptors might be involved in the neuroinflammatory
process that develops in some forms of neurodegeneration.
Collectively, CB1 is expressed at low levels in glial cells
and CB2 can be considered the inducible receptor, thus ex-
plaining their different discriminative, functional importance
in glia.
Endocannabinoids
Different types of glial cells can produce and release en-
docannabinoids (EC) [40,46] and during traumatic brain in-
jury and neuroinflammatory diseases, an increased produc-
tion can be observed [47,48]. The cellular source of EC pro-
duced during neuroinflammation is uncertain, although cul-
tured mouse microglial cells have been proved to produce
about 20-fold more EC than neurons and astrocytes [23].
As in neurons, in glia too EC do not accumulate but are
actively secreted “on demand” immediately after they are
synthesized from their membrane precursors by lipase en-
zymes. They activate cannabinoid receptors and are taken up
with a mechanism still under discussion, and subjected to
enzymatic hydrolysis. Anandamide (AEA), one of the most
important EC [49-51], behaves as a partial agonist in stimu-
lating cannabinoid receptors and, like the majority of EC, it
is hydrolyzed mainly by the fatty acid amide hydrolase
(FAAH). In contrast, 2-arachidonoylglycerol (2-AG), the EC
found in the largest amounts in the CNS, which behaves as a
full agonist at cannabinoid receptors, seems to be mainly
inactivated in vivo by an enzyme called monoacylglyceride
lipase (MAGL) [52,53], whereas FAAH seems to have a
secondary role in its degradation, as indicated by the un-
changed brain levels of 2-AG in wild-type and FAAH -/-
mice [54].
Degrading Enzymes
FAAH is an integral membrane protein responsible for
the breakdown of several lipophilic compounds including the
endocannabinoid AEA. Mainly it mediates the termination of
Massi et al.
the AEA signal though it also appears to be involved in ces-
sation of the 2-AG signal [55,56]. This enzyme was reported
by Romero et al. [57] in human brain neurons and in cortical
and basal ganglia astrocytes, overlapping significantly with
CB1 receptors, mainly in areas related to motor control and
memory. Surprisingly, Tsou et al. [58] and Egertova et al.
[59] did not find any expression of FAAH in glial cells in rat
brain, probably because of species differences in its expres-
sion pattern.
Benito et al. [29] obtained very interesting data in post-
mortem brain tissues from Alzheimer patients. FAAH pro-
tein and activity and CB2 receptors were selectively overex-
pressed in astrocytes and microglia, respectively, associated
with the
A-rich neuritic plaques seen in this disease,
whereas they seemed to be present only at a very low level,
or were absent, in glial cells of healthy subjects. Since AEA
is a substrate for FAAH and is converted into arachidonic
acid, the massive presence and activity of FAAH associated
with the Alzheimer plaques might be a significant source of
arachidonic acid and related pro-inflammatory substances in
the vicinity of these plaques, with deleterious effects. Thus,
inhibition of FAAH activity has been proposed as a means of
preventing the inflammatory process associated with ßA
deposition in AD.
The pattern of FAAH selective overexpression may be
identical in other neuroinflammatory processes. In macaques
with SIVE strong FAAH immunoreactivity was found in
astrocytes near the inflammatory infiltrates [41]. Changes
similar to those in SIVE were also described in samples from
patients with MS [42]. Nunez et al. [31] described increases
in CB2 and FAAH enzyme in ßA plaque–related microglia
and astroglia, respectively, in DS, sometimes referred to as a
human model of Alzheimer-like ßA deposition. Thus, as
suggested by the findings in AD, MS, SIVE and DS, the
overexpression of FAAH in several neuroinflammatory con-
ditions may have a specific pathogenic role.
Massi et al. [60] reported that in glioma tissues and in
U87 human glioma cells there was a striking increase of
FAAH activity, not paralleled by any increase in FAAH con-
tent, after in vivo and in vitro exposure to the non-psycho-
active, antitumoral cannabinoid compound cannabidiol
(CBD). One of the products of FAAH activity is arachidonic
acid, which can behave as an pro-inflammatory/apoptotic
agent. Thus, the upregulation of FAAH by CBD in gliomas
might conceivably be one mechanism through which can-
nabinoids influence tumor growth.
Although there is ample information on FAAH, much
less is known about MAGL and 2-AG hydrolysis. Muccioli
et al. [61] provided the first evidence of an unspecified
MAGL activity expressed by microglial cells. They found
that the mouse microglial cell line BV-2 did not express
MAGL mRNA but there was efficient 2-AG hydrolysis.
URB597, a synthetic FAAH inhibitor, halved this hydrolysis
and the remaining activity was blocked by classic MAGL
inhibitors such as URB602. These findings, confirmed in
mouse primary microglia in culture, suggested there was
some previously unknown MAGL activity that controls 2-
AG levels in microglia. Although MAGL expressed in neu-
rons is equally distributed between the cytosolic, mitochon-
Expression and Function of the Endocannabinoid System in Glial Cells
drial, and nuclear fractions, the activity expressed by BV-2
cells was richer in mitochondrial and nuclear fractions.
Effect of (Endo)Cannabinoids on Glial Cell Reactivity
and Function
EC act on glial cells modulating their important immu-
nological function and promoting biochemical signals ending
in a pro-survival fate for these cells. It is well known that
neuroinflammatory, cytotoxic and ischemic events induce
complex and dynamic changes in glial cell phenotype, with
morphological changes and release of compounds such as
ROS, NO and proinflammatory cytokines.
Here, we try to summarize the few data present in the
literature on the ability of EC to modulate glial cells reactiv-
ity and/or their specific production of cytokines.
Astrocytes
Primary cultures of neonatal mouse cortical astrocytes
stimulated with lipopolysaccharide or Theiler's murine en-
cephalomyelitis virus (TMEV) (which elicits a mouse model
of MS) released significant amounts of nitrites and TNF-
alpha into the culture medium [62]. AEA inhibited the re-
lease of these proinflammatory molecules in a dose-
dependent manner, suggesting ECs play a clear part in im-
mune reactivity in the brain [62]. AEA also enhanced IL-6
release from astrocytes infected with TMEV; this pleiotropic
cytokine is involved in the regulation of immunological re-
sponses, with some neuroprotective effects [63].
Ortega-Gutierrez et al. [64] reported that AEA in astro-
cytes reduced NO release, iNOS expression and the produc-
tion of the proinflammatory cytokines TNF-alpha and IL-
1beta, and these effects were mimicked by the synthetic can-
nabinoid compound HU210.
Microglia
Puffenbarger et al. [65] found that exposure of neonatal
rat cortical microglial cells to THC, methandamide or AEA
reduced the amounts of LPS-induced mRNAs for the proin-
flammatory or immunoregulatory cytokines independently
by either CB1 or CB2 cannabinoid receptors stimulation. It
was also shown that UCM707, a selective inhibitor of the
cellular reuptake of AEA, improved motor function in
TMEV-infected mice and diminished neuroinflammation.
These effects run in parallel to the decrease in the microglial
reactivity and the strong reduction in MHC class II antigen
expression on these cells [66]. Primary cultures of rat micro-
glia activated by LPS release TNF-alpha cytokine, like as-
trocytes, and this was inhibited by AEA, 2-AG, CP-55940
and WIN-55,212-2 [67]. Moreover, cannabinoids can oppose
microglial CD40 expression, an important marker of micro-
glial activation, contributing to the start and/or progression
of microglial production of proinflammatory mediators [68].
Interestingly, in glial cells EC can also enhance the release of
the anti-inflammatory cytokine IL-1ra [69]. Taken together,
these results suggest that EC reduce neurotoxicity and have
significant immunoregulatory role in glial cells.
Eljaschewitsch et al. [70] reported that AEA induced
histone H3 phosphorylation, expression of the mitogen-
activated protein kinase phosphatase-1 (MKP-1), and subse-
quent ERK-1/2 dephosphorylation in activated but not rest-
Current Pharmaceutical Design, 2008, Vol. 14, No. 23 2293
ing microglial cells, which in turn abolishes NO release, fi-
nally resulting in neuroprotection. The authors concluded
that release of EC in injured brain tissue might constitute a
signal to glial cells to limit neuronal degeneration, thus re-
ducing the extent of the inflammatory response.
Glial Cells Migration
An apparent contradictory result was reported concerning
the ability of EC to trigger rather than limit glial cells migra-
tion . As a matter of fact, Walter [46] showed that astrocytes
in vitro produce AEA and many other acylethanolamines
under Ca++-dependent stimuli. Microglia were also reported
to produce EC at higher levels than neurons [23]. Thus, it is
conceivable that during neuroinflammation activated micro-
glia produce the majority of EC that accumulate at the lesion
site. It has been reported that 2-AG triggers microglial mi-
gration, with high potency and efficacy, probably as a signal
to recruit microglia to lesioned sites [23]. The same group
also showed that palmitoylethanolamide (PEA), an endoge-
nous structural analog of anandamide that acts as an “entou-
rage” compound for AEA [71], which increases after focal
cerebral ischemia in mice, can stimulate microglial cell mi-
gration and potentiate AEA-induced microglial cell motility
[72]. Since this increase in cell motility is essential for the
propagation of neuroinflammation but also to control at the
onset the correct response to stop an inflammatory insult, the
authors concluded by suggesting pharmacological control of
PEA and AEA signaling to influence cell migration.
In summary, these studies suggest that at least some of
the cannabinoids’ protective effects may be due to a complex
relationship between inhibition/stimulation of the release of
some proinflammatory/anti-inflammatory mediators from
glia and the modulation of microglial migration/activation.
The ability of EC to modulate specific cytokines might sug-
gest that like in periphery [73] EC could alter the equilibrium
between Th1 and Th2 immune response, but the scarce and
partial data available at present do not permit to translate this
modulation to CNS.
Cannabinoids Influence Glial Cell Survival
Another mechanism through which cannabinoids can re-
gulate glial cells is their ability to affect the cells’ fate, influ-
encing their survival.
Astrocytes
Cannabinoids protect glial cells from apoptosis in vivo
and in vitro (Fig. 1). THC, WIN55,212-2 and HU210 protect
primary astrocytes in culture from ceramide-induced apopto-
sis through activation of the PI3K/PKB pathway and ERK
signal; in vivo WIN55,212-2 prevented the loss of astrocytes
induced by focal injection of ceramide into the hippocampus
[74]. These results are important, because exposure to proin-
flammatory cytokines [75] or saturated fatty acid [76] can
increase ceramide generation in astrocytes in conditions of
brain disease or injury.
Further studies have indicated that THC and WIN55,212-
2 protect astrocytes from oxidative stress damage since pro-
longed incubation with these two compounds prevented
H2O2-induced loss of viability through a mechanism mainly
involving CB1 cannabinoid receptor stimulation [77]. It has
2294 Current Pharmaceutical Design, 2008, Vol. 14, No. 23
been demonstrated that the stress state in the cells induced by
serum deprivation leads to marked changes in the astrocytes’
gene expression profile, in particular in the genes involved in
protection against oxidative stress and in the control of cell
survival as the stress-regulated protein p8 [78].
Microglia
Stimulated rat RTMGL1 microglial cells mainly produce
the endocannabinoid 2-AG, expressing the CB2 receptor at
an easily detectable level and the CB1 receptor in smaller
amounts [40]. When added exogenously to the microglial
cell culture, 2-AG but not AEA increases cell proliferation,
probably by boosting ERK activity, suggesting that micro-
glia produce and release 2-AG as an autacoid co-stimulatory
molecule in stimulated cells. Since microglial proliferation
has been sometimes linked to many chronic neurodegenera-
tive pathologies, pharmacological manipulation of the ECs
might well have important therapeutic implications.
Oligodendrocytes
Oligodendrocytes are highly susceptible to hypoxia [79],
oxidative stress [80], and humoral and cellular immune-
mediated attack [81]. Molina-Holgado et al. [15] showed
that cannabinoids (HU-210, WIN55,212-2) protected oli-
godendrocyte progenitors expressing CB1 and CB2 receptors
from apoptosis induced by deprivation of trophic support,
through a mechanism dependent on the PI3K/Akt signaling
pathway. This is an important finding considering that mye-
lin disorders affecting oligodendrocytes are among the major
CNS pathologies. Since oligodendrocyte progenitors proba-
bly exist for some sort of repair strategies, a prosurvival sig-
nal for these cells, provided by ECs – together with others of
apoptotic significance such as trophic deprivation and unfa-
vorable conditions – might be very important in the ECs’
protective role in demyelinating diseases [15].
Neural Progenitors
Aguado et al. [82] provided evidence that in neural pro-
genitor (NP) cells the ECs can act as an instructive proglio-
genic signal. Exposure of NP cells to cannabinoids regulates
the cell differentiation toward the astroglial lineage. Astro-
glial differentiation was lower in CB1-deficient mice and
higher in FAAH-deficient mice. Thus, the ECs’s effects on
NP cell proliferation and differentiation may be important
for brain development and plasticity and for the development
of the fundamental glia-neurons network. These findings
merit further investigation in view of the importance of the
relationship between correct CNS development and neu-
rodegenerative disorders.
In summary, EC can stimulate glial cell proliferation and
survival, through a mechanism to control the correct expan-
sion and preservation of this population, fundamental in sus-
taining proper neuronal function (Fig. 1).
The Endocannabinoid System in Glia-Derived Brain
Tumors
The term glioma broadly classifies primary neoplastic
tissue of glial origin. Glioma cancer can be considered as a
particular and extreme form of neuroinflammatory/neurode-
generative state, and the presence and expression of cannabi-
noid receptors in tumors from glial cells has been investi-
Massi et al.
gated. Like primary glial cells, gliomas have all the compo-
nents of the ECs, such as CB1 and/or CB2 receptors, the
ability to synthesize one or more EC, AEA membrane trans-
porter, the enzyme FAAH which inactivates EC [83,84].
This has suggested that ECs may be involved in controlling
abnormal cell proliferation [85].
(Endo)Cannabinoids and Glioma Cell Proliferation
The effect of natural and synthetic cannabinoids such as
THC, CBD, or JWH-133 have been investigated in glioma,
and they appear to have potent antiproliferative action
[83,84]. In contrast, the effects of EC in this type of tumor
have not been extensively investigated, although this might
offer a potential alternative approach for the development of
harmless anti-cancer drugs. It has been demonstrated that
gliomas, like normal glia, can release EC. Petersen et al. [86]
reported that human glioblastoma had significantly higher
levels of N-acylethanolamines, including AEA and 2-AG,
than normal brain tissue. Maccarrone et al. [87] described a
pro-apoptotic effect of AEA in C6 rat glioma cells due to
activation of vanilloid receptors. Jacobsson et al. [88]
showed that in rat C6 glioma cells AEA had an antiprolifera-
tive effect associated with combined activation of cannabi-
noid and vanilloid receptors, with potency similar to that
reported for 2-AG. Another endocannabinoid, stearoyletha-
nolamide (SEA), found in human brain in amounts compara-
ble to AEA, showed pro-apoptotic activity in a glioma cell
line [89]. Contassot et al. [90] found that human glioma cell
lines, either established for a very long time (U87 and U251)
or derived from a tumor biopsy (Ge227 and Ge258), were
efficiently killed by AEA. Thus, the meaning and the func-
tion of the EC synthesis and release in glioma and in normal
glia still represents matter of debate, since it appears that
while EC have proliferative/prosurvival effects in normal
glial cells, in glioma cells they induce death as opposite ef-
fect [83].
EC inhibit glioma cell proliferation through multiple
mechanisms [83,84]. Apoptosis of the cells can be mediated
by an acute increase of sphingomyelin breakdown and long-
term activation of JNK and p38 MAPK or raising intracellu-
lar calcium (Fig. 2). Moreover, CBD, a non-psychotropic
cannabinoid, is capable of killing glioma cells by triggering
prompt production of ROS and marked depletion of GSH
content [91,92]. Thus, the oxidation pathway is presumably
very important and might be one of the main mechanisms of
cannabinoid action in cancer cells (Fig. 2).
In addition, substantial evidence has been collected sup-
porting a fundamental role for lipoxygenase (LOX)- and
cyclooxygenase (COX)-catalyzed arachidonic acid metabo-
lism in cancer. The isoenzymes most involved in the control
of cell growth and death within the central nervous system
are 5-LOX [93] and COX-2 [94]. The endocannabinoid SEA
induces apoptosis in C6 glioma cells through activation of
COX and LOX, without involving MAPK cascades [89].
Moreover, when nude mice bearing subcutaneous gliomas
were treated in vivo with CBD the activity and content of 5-
LOX were significantly reduced in tumor tissues [60]. These
data provide a further demonstration of alternative intracellu-
lar targets that could be largely influenced by cannabinoids,
contributing to their antitumoral effect.
Expression and Function of the Endocannabinoid System in Glial Cells Current Pharmaceutical Design, 2008, Vol. 14, No. 23 2295

Fig. (2) . (Endo)cannabinoids and inhibition of glioma cell proliferation.

Natural, synthetic and endogenous cannabinoids exert their anti-tumoral effects affecting various cellular pathways by binding the specific
CB1 and CB2 Gi/o–protein coupled receptors (or unknown CBn receptor). CB receptor signal triggers the inhibition of the activity of adeny-
late cyclase (AC). The decrease in the cAMP levels causes a consequent inhibition of protein kinase (PKA) pathway determining an inhibi-
tion of tumor cell cycle. Cannabinoid receptors can also promote Ca++ release from intracellular stores, thus controlling cell fate. The stimu-
lation of cannabinoid receptors may lead to an increase in ceramide levels mainly through a synthesis de novo. Ceramide accumulation can
cause an upregulation of p8 and in turn of the Endoplasmic Reticulum stress-related genes ATF-4, CHOP and TRB3. Ceramide also activates
JNK and p38. The activation of these pathways determines an inhibition of cell proliferation. In addition, cannabinoids can trigger reactive
oxygen species (ROS), elevation of intracellular calcium, modulation of the arachidonate cascade, dissipation of mitochondrial potential
(
), reduction of GSH, inhibition of LOX activity and content, ultimately leading to cell death. The crosstalk between the different path-
ways and many molecular details have been omitted for simplification.

Selectivity of Cannabinoid Action on Tumor Cells
The ability of EC to induce either cell survival or death,
depending on the cells concerned - in this case respectively
primary glial cells or glioma cells - is very interesting. It
appears that cannabinoids can selectively kill tumor cells
without affecting their non-transformed counterparts. This
apparent selective effect would be of enormous clinical im-
portance.
The THC-induced cell death observed in rat C6 glioma
cells did not occur in non-transformed primary astrocytes
[95]. In vivo cannabinoids inhibit, and to some extent eradi-
cate, C6 glioma tumors, with no discernable damage to sur-
rounding healthy tissue [96].
The difference in sensitivity might be due to differences
in signal transduction events downstream of the cell surface
and/or different redox state-cellular features of the tumoral
cells compared to normal cells. The first hypothesis is sup-
ported by Carracedo’s papers [77,78] which report that can-
nabinoid treatment did not trigger ceramide accumulation,
upregulation of p8 or other ER stress-related genes in non-
transformed astrocytes. The oxidative hypothesis is consis-
tent with the finding that oxidative metabolism and associ-
ated sensitivity to apoptosis in tumor cells differ from nor-
mal cells [97,98].
This selective property is shared by the non-psychotropic
CBD, that reduced glioma cell growth with no apparent tox-
icity on non-transformed cell counterparts, as reported by
Massi et al. [92]. Also Duntsch et al. [99] showed the selec-
tive efficacy of the CB2 cannabinoid agonist KM-233 in
U87 glioma cells.
To summarize, it can be proposed that this dual property
might reflect the differences in the features of normal and
transformed cells. The opposite regulation of CB1 and CB2
expression in glioma and normal glial cells might explain
their different sensitivity to cannabinoids but this does not
seem to be an univocal explanation because, besides the re-
ceptor-independent effect of cannabinoids in glioma cell
death [91,92,95], glioma cells and glial cells possess both
cannabinoid receptors [22,43].
CONCLUSION
Data reported in the present review clearly support the
hypothesis that ECs could play a crucial role in regulating
glia activity during physiological and pathological conditions
2296 Current Pharmaceutical Design, 2008, Vol. 14, No. 23
of the brain. Glial cells produce endocannabinoids, express
functional CB1/CB2 cannabinoid receptors, and degrade
endocannabinoids. In addition, during pathological condition
of the brain, an upregulation of endocannabinoid production
as well as of the other components of ECs has been found.
These responses are likely part of an endogenous mechanism
of defense against a variety of brain-damaging insults,
probably representing a mechanism to maintain brain ho-
meostasis and to oppose the injury associated with condi-
tions of inflammation, trauma, excitotoxicity, infection and
other neuropathological stimuli (Fig. 1). Thus, the enhance-
ment of the endocannabinoid signaling through the inhibition
of the enzymes degrading endocannabinoids or targeting
cannabinoid receptors by using exogenous/endogenous can-
nabinoids could constitute an innovative therapeutic ap-
proach for neurodegenerative diseases.
Conversely, EC have been shown to be potent and selec-
tive antiproliferative and proapototic agents in transformed
glial cells acting by multiple cellular mechanisms not always
depending by cannabinoid receptor stimulation. Data about
the role of ECs in glioma growth seem to indicate that this
system specifically affects many of the function that are spe-
cific and essential in cancer cell growth inducing a valuable
proapoptotic signaling (Fig. 2).
Taken together, the data reviewed in this paper indicate
that manipulation of ECs can offer a multi-faceted approach
for the treatment of some brain pathologies by providing
either useful neuroprotection or, as double side of the coin,
limiting the glia-derived tumors growth.
REFERENCES
References 100-102 are related articles recently published.
[1] Garden GA, Moller T. Microglia biology in health and disease. J
Neuroimmune Pharmacol 2006; 1: 127-37.
[2] Dheen ST, Kaur C, Ling EA. Microglial activation and its implica-
tions in the brain diseases. Curr Med Chem 2007; 14(11): 1189-97.
[3] Hanisch UK, Kettenmann H. Microglia: active sensor and versatile
effector cells in the normal and pathologic brain. Nat Neurosci
2007; 10: 1387-94.
[4] Vesce S, Bezzi P, Volterra A. Synaptic transmission with the glia.
News Physiol Sci 2001; 16: 178–84.
[5] Bohn MC. Motoneurons crave glial cell line-derived neurotrophic
factor. Exp Neurol 2004; 190: 263–75.
[6] Fellin T, Carmignoto G. Neuron-to-astrocytes signalling in the
brain represents a distinct multifunctional unit. J Physiol 2004; 559:
3-15.
[7] Pellerin L, Bouzier-Sore AK, Aubert A, Serres S, Merle M, Costa-
lat R, et al. Magistretti PJ Activity-dependent regulation of energy
metabolism by astrocytes: an update. Glia 2007; 55: 1251-62.
[8] Rubin LL, Staddon JM. The cell biology of the blood-brain barrier.
Annu Rev Neurosci 1999; 22: 11-28.
[9] Rodriguez JJ, Mackie K, Pickel VM. Ultrastructural localization of
the CB1 cannabinoid receptor in mu-opioid receptor patches of the
rat Caudate putamen nucleus. J Neurosci 2001; 21: 823-33.
[10] Moldrich G, Wenger T. Localization of the CB1 cannabinoid re-
ceptor in the rat brain. An immunohistochemical study. Peptides
2000; 21: 1735-42.
[11] Salio C, Doly S, Fischer J, Franzoni MF, Conrath M. Neuronal and
astrocytic localization of the cannabinoid receptor-1 in the dorsal
horn of the rat spinal cord. Neurosci Lett 2002; 329: 13-6.
[12] Bouaboula M, Bourrié B, Rinaldi-Carmona M, Shire D, Le Fur G,
Casellas P. Stimulation of cannabinoid receptor CB1 induces krox-
24 expression in human astrocytoma cells. J Biol Chem 1995; 270:
13973-80.
Massi et al.
[13] Sánchez C, Galve-Roperh I, Canova C, Brachet P, Guzmán M.
Delta9-tetrahydrocannabinol induces apoptosis in C6 glioma cells.
FEBS Lett 1998a; 436: 6-10.
[14] Abood ME, Rizvi G, Sallapudi N, McAllister SD. Activation of the
CB1 cannabinoid receptor protects cultured mouse spinal neurons
against excitotoxicity. Neurosci Lett 2001; 309: 197-201.
[15] Molina-Holgado E, Vela JM, Arévalo-Martín A, Almazán G, Mo-
lina-Holgado F, Borrell J, et al. Cannabinoids promote oligoden-
drocyte progenitor survival: involvement of cannabinoid receptors
and phosphatidylinositol-3 kinase/Akt signaling. J Neurosci 2002;
22: 9742-53.
[16] Navarrete M, Araque A. Endocannabinoids mediate neuron-astro-
cyte communication. Neuron 2008; 57: 883-93.
[17] Walter L, Stella N. Endothelin-1 increases 2-arachidonoyl glycerol
(2-AG) production in astrocytes. Glia 2003; 44: 85-90.
[18] Sagan S, Venance L, Torrens Y, Cordier J, Glowinski J, Giaume C.
Anandamide and WIN 55212-2 inhibit cyclic AMP formation
through G-protein-coupled receptors distinct from CB1 cannabi-
noid receptors in cultured astrocytes. Eur J Neurosci 1999; 11: 691-
9.
[19] Szabo B, Schlicker E. Effects of cannabinoids on neurotransmis-
sion. Handb Exp Pharmacol 2005; 168: 327-65.
[20] Sánchez C, Galve-Roperh I, Rueda D, Guzmán M. Involvement of
sphingomyelin hydrolysis and the mitogen-activated protein kinase
cascade in the Delta9-tetrahydrocannabinol-induced stimulation of
glucose metabolism in primary astrocytes. Mol Pharmacol 1998b;
54: 834-43.
[21] Blázquez C, Sánchez C, Daza A, Galve-Roperh I, Guzmán M. The
stimulation of ketogenesis by cannabinoids in cultured astrocytes
defines carnitine palmitoyltransferase I as a new ceramide-
activated enzyme. J Neurochem 1999; 72: 1759-68.
[22] Maresz K, Carrier EJ, Ponomarev ED, Hillard CJ, Dittel BN.
Modulation of the cannabinoid CB2 receptor in microglial cells in
response to inflammatory stimuli. J Neurochem 2005; 95: 437-45.
[23] Walter L, Franklin A, Witting A, Wade C, Xie Y, Kunos G, et al.
Nonpsychotropic cannabinoid receptors regulate microglial cell
migration. J Neurosci 2003; 23: 1398-405.
[24] Waksman Y, Olson JM, Carlisle SJ, Cabral GA. The central can-
nabinoid receptor (CB1) mediates inhibition of nitric oxide produc-
tion by rat microglial cells. J Pharmacol Exp Ther 1999; 288: 1357-
66.
[25] Derocq JM, Ségui M, Marchand J, Le Fur G, Casellas P. Cannabi-
noids enhance human B-cell growth at low nanomolar concentra-
tions. FEBS Lett 1995; 369: 177-82.
[26] McCoy KL, Matveyeva M, Carlisle SJ, Cabral GA. Cannabinoid
inhibition of the processing of intact lysozyme by macrophages:
evidence for CB2 receptor participation. J Pharmacol Exp Ther
1999; 289: 1620-5.
[27] Sugiura T, Kondo S, Kishimoto S, Miyashita T, Nakane S, Kodaka
T, et al. Evidence that 2-arachidonoylglycerol but not N-palmitoyl-
ethanolamine or anandamide is the physiological ligand for the
cannabinoid CB2 receptor. Comparison of the agonistic activities
of various cannabinoid receptor ligands in HL-60 cells. J Biol
Chem 2000; 275: 605-12.
[28] Carlisle SJ, Marciano-Cabral F, Staab A, Ludwick C, Cabral GA.
Differential expression of the CB2 cannabinoid receptor by rodent
macrophages and macrophage-like cells in relation to cell activa-
tion. Int Immunopharmacol 2002; 2: 69-82.
[29] Benito C, Núñez E, Tolón RM, Carrier EJ, Rábano A, Hillard CJ,
et al. Cannabinoid CB2 receptors and fatty acid amide hydrolase
are selectively overexpressed in neuritic plaque-associated glia in
Alzheimer's disease brains. J Neurosci 2003; 23: 11136-41.
[30] Ramírez BG, Blázquez C, Gómez del Pulgar T, Guzmán M, de
Ceballos ML. Prevention of Alzheimer's disease pathology by can-
nabinoids: neuroprotection mediated by blockade of microglial ac-
tivation. J Neurosci 2005; 25: 1904-13.
[31] Núñez E, Benito C, Tolón RM, Hillard CJ, Griffin WS, Romero J.
Glial expression of cannabinoid CB(2) receptors and fatty acid am-
ide hydrolase are beta amyloid-linked events in Down's syndrome.
Neuroscience 2008; 151: 104-10.
[32] Onaivi ES, Ishiguro H, Gong JP, Patel S, Meozzi PA, Myers L,
et al. Brain neuronal CB2 cannabinoid receptors in drug abuse and
depression: from mice to human subjects. PLoS ONE 2008; 3:
1640.
Expression and Function of the Endocannabinoid System in Glial Cells
[33] Van Sickle MD, Duncan M, Kingsley PJ, Mouihate A, Urbani P,
Mackie K, et al. Identification and functional characterization of
brainstem cannabinoid CB2 receptors. Science 2005; 310: 329-32.
[34] Gong JP, Onaivi ES, Ishiguro H, Liu QR, Tagliaferro PA, Brusco
A, et al. Cannabinoid CB2 receptors: immunohistochemical local-
ization in rat brain. Brain Res 2006; 1071: 10-23.
[35] Molina-Holgado F, Pinteaux E, Heenan L, Moore JD, Rothwell NJ,
Gibson RM. Neuroprotective effects of the synthetic cannabinoid
HU-210 in primary cortical neurons are mediated by phosphatidy-
linositol 3-kinase/AKT signaling. Mol Cell Neurosci 2005; 28:
189-94.
[36] Ashton JC, Friberg D, Darlington CL, Smith PF. Expression of the
cannabinoid CB2 receptor in the rat cerebellum: an immunohisto-
chemical study. Neurosci Lett 2006; 396: 113-6.
[37] Golech SA, McCarron RM, Chen Y, Bembry J, Lenz F, Mechou-
lam R, et al. Human brain endothelium: coexpression and function
of vanilloid and endocannabinoid receptors. Brain Res Mol Brain
Res 2004; 132: 87-92.
[38] Correa F, Mestre L, Molina-Holgado E, Arévalo-Martín A, Doca-
gne F, Romero E, et al. The role of cannabinoid system on immune
modulation: therapeutic implications on CNS inflammation. Mini
Rev Med Chem 2005; 5: 671-5.
[39] Núñez E, Benito C, Pazos MR, Barbachano A, Fajardo O, Gonzá-
lez S, et al. Cannabinoid CB2 receptors are expressed by perivascu-
lar microglial cells in the human brain: an immunohistochemical
study. Synapse 2004; 53: 208-13.
[40] Carrier EJ, Kearn CS, Barkmeier AJ, Breese NM, Yang W, Nithi-
patikom K, et al. Cultured rat microglial cells synthesize the endo-
cannabinoid 2-arachidonylglycerol, which increases proliferation
via a CB2 receptor-dependent mechanism. Mol Pharmacol 2004;
65: 999-1007.
[41] Benito C, Kim WK, Chavarría I, Hillard CJ, Mackie K, Tolón RM,
et al. A glial endogenous cannabinoid system is upregulated in the
brains of macaques with simian immunodeficiency virus-induced
encephalitis. J Neurosci 2005; 25: 2530-6.
[42] Benito C, Romero JP, Tolón RM, Clemente D, Docagne F, Hillard
CJ, et al. Cannabinoid CB1 and CB2 receptors and fatty acid amide
hydrolase are specific markers of plaque cell subtypes in human
multiple sclerosis. J Neurosci 2007; 27: 2396-402.
[43] Cabral GA, Marciano-Cabral F. Cannabinoid receptors in microglia
of the central nervous system: immune functional relevance. J
Leukoc Biol 2005; 78: 1192-7.
[44] Pazos MR, Núñez E, Benito C, Tolón RM, Romero J. Functional
neuroanatomy of the endocannabinoid system. Pharmacol Biochem
Behav 2005; 81: 239-47.
[45] Glass M, Dragunow M, Faull RL. Cannabinoid receptors in the
human brain: a detailed anatomical and quantitative autoradio-
graphic study in the fetal, neonatal and adult human brain. Neuro-
science 1997; 77: 299-318.
[46] Walter L, Franklin A, Witting A, Moller T, Stella N. Astrocytes in
culture produce anandamide and other acylethanolamides. J Biol
Chem 2002; 277: 20869-76.
[47] Panikashvili D, Simeonidou C, Ben-Shabat S, Hanus L, Breuer A,
Mechoulam R, et al. An endogenous cannabinoid (2-AG) is neuro-
protective after brain injury. Nature 2001; 413: 527-31.
[48] Baker D, Pryce G, Croxford JL, Brown P, Pertwee RG, Makriyan-
nis A, et al. Endocannabinoids control spasticity in a multiple scle-
rosis model. FASEB J 2001; 15: 300-2.
[49] Sun YX, Tsuboi K, Okamoto Y, Tonai T, Murakami M, Kudo I,
et al. Biosynthesis of anandamide and N-palmitoylethanolamine by
sequential actions of phospholipase A2 and lysophospholipase D.
Biochem J 2004; 380: 749-56.
[50] Liu J, Wang L, Harvey-White J, Osei-Hyiaman D, Razdan R, Gong
Q, et al. A biosynthetic pathway for anandamide. Proc Natl Acad
Sci USA 2006; 103: 13345-50.
[51] Simon GM, Cravatt BF. Endocannabinoid biosynthesis proceeding
through glycerophospho-N-acyl ethanolamine and a role for al-
pha/beta-hydrolase 4 in this pathway. J Biol Chem 2006; 281:
26465-72.
[52] Dinh TP, Freund TF, Piomelli D. A role for monoglyceride lipase
in 2-arachidonoylglycerol inactivation. Chem Phys Lipids 2002;
121: 149-58.
[53] Saario SM, Savinainen JR, Laitinen JT, Järvinen T, Niemi R.
Monoglyceride lipase-like enzymatic activity is responsible for hy-
drolysis of 2-arachidonoylglycerol in rat cerebellar membranes.
Biochem Pharmacol 2004; 67: 1381-7.
Current Pharmaceutical Design, 2008, Vol. 14, No. 23 2297
[54] Lichtman AH, Hawkins EG, Griffin G, Cravatt BF. Pharmacologi-
cal activity of fatty acid amides is regulated, but not mediated, by
fatty acid amide hydrolase in vivo. J Pharmacol Exp Ther 2002;
302: 73-9.
[55] Deutsch DG, Glaser ST, Howell JM, Kunz JS, Puffenbarger RA,
Hillard CJ, et al. The cellular uptake of anandamide is coupled to
its breakdown by fatty-acid amide hydrolase. J Biol Chem 2001;
276: 6967-73.
[56] Deutsch DG, Ueda N, Yamamoto S. The fatty acid amide hydro-
lase (FAAH). Prostaglandins Leukot Essent Fatty Acids 2002; 66:
201-10.
[57] Romero J, Lastres-Becker I, de Miguel R, Berrendero F, Ramos
JA, Fernández-Ruiz J. The endogenous cannabinoid system and the
basal ganglia. biochemical, pharmacological, and therapeutic as-
pects. Pharmacol Ther 2002; 95: 137-52.
[58] Tsou K, Nogueron MI, Muthian S, Sañudo-Pena MC, Hillard CJ,
Deutsch DG, et al. Fatty acid amide hydrolase is located preferen-
tially in large neurons in the rat central nervous system as revealed
by immunohistochemistry. Neurosci Lett 1998; 254: 137-40.
[59] Egertová M, Giang DK, Cravatt BF, Elphick MR. A new perspec-
tive on cannabinoid signalling: complementary localization of fatty
acid amide hydrolase and the CB1 receptor in rat brain. Proc Biol
Sci 1998; 265: 2081-5.
[60] Massi P, Valenti M, Vaccani A, Gasperi V, Perletti G, Marras E,
et al. 5-Lipoxygenase and anandamide hydrolase (FAAH) mediate
the antitumor activity of cannabidiol, a non-psychoactive cannabi-
noid. J Neurochem 2008; 104: 1091-100.
[61] Muccioli GG, Xu C, Odah E, Cudaback E, Cisneros JA, Lambert
DM, et al. Identification of a novel endocannabinoid-hydrolyzing
enzyme expressed by microglial cells. J Neurosci 2007; 27: 2883-9.
[62] Molina-Holgado F, Lledó A, Guaza C. Anandamide suppresses
nitric oxide and TNF-alpha responses to Theiler's virus or endo-
toxin in astrocytes. Neuroreport 1997; 8: 1929-33.
[63] Molina-Holgado F, Molina-Holgado E, Guaza C. The endogenous
cannabinoid anandamide potentiates interleukin-6 production by
astrocytes infected with Theiler's murine encephalomyelitis virus
by a receptor-mediated pathway. FEBS Lett 1998; 433: 139-42.
[64] Ortega-Gutiérrez S, Molina-Holgado E, Guaza C. Effect of anan-
damide uptake inhibition in the production of nitric oxide and in
the release of cytokines in astrocyte cultures. Glia 2005; 52: 163-8.
[65] Puffenbarger RA, Boothe AC, Cabral GA. Cannabinoids inhibit
LPS-inducible cytokine mRNA expression in rat microglial cells.
Glia 2000; 29: 58-69.
[66] Ortega-Gutiérrez S, Molina-Holgado E, Arévalo-Martín A, Correa
F, Viso A, et al. Activation of the endocannabinoid system as thera-
peutic approach in a murine model of multiple sclerosis. FASEB J
2005; 19: 1338-40.
[67] Facchinetti F, Del Giudice E, Furegato S, Passarotto M, Leon A.
Cannabinoids ablate release of TNFalpha in rat microglial cells
stimulated with lypopolysaccharide. Glia 2003; 41: 161-8.
[68] Ehrhart J, Obregon D, Mori T, Hou H, Sun N, Bai Y, et al. Stimu-
lation of cannabinoid receptor 2 (CB2) suppresses microglial acti-
vation. J Neuroinflamm 2005; 2: 29.
[69] Molina-Holgado F, Pinteaux E, Moore JD, Molina-Holgado E,
Guaza C, Gibson RM, et al. Endogenous interleukin-1 receptor an-
tagonist mediates anti-inflammatory and neuroprotective actions of
cannabinoids in neurons and glia. J Neurosci 2003; 23: 6470-4.
[70] Eljaschewitsch E, Witting A, Mawrin C, Lee T, Schmidt PM, Wolf
S, et al. The endocannabinoid anandamide protects neurons during
CNS inflammation by induction of MKP-1 in microglial cells.
Neuron 2006; 49: 67-79.
[71] De Petrocellis L, Bisogno T, Ligresti A, Bifulco M, Melck D, Di
Marzo V. Effect on cancer cell proliferation of palmitoylethanola-
mide, a fatty acid amide interacting with both the cannabinoid and
vanilloid signalling systems. Fundam Clin Pharmacol 2002; 16:
297-302.
[72] Franklin A, Parmentier-Batteur S, Walter L, Greenberg DA, Stella
N. Palmitoylethanolamide increases after focal cerebral ischemia
and potentiates microglial cell motility. J Neurosci 2003; 23: 7767-
75.
[73] Massi P, Vaccani A, Parolaro D. Cannabinoids, immune system
and cytokine network. Curr Pharm Des 2006; 12: 3135-46.
[74] Gómez Del Pulgar T, De Ceballos ML, Guzmán M, Velasco G.
Cannabinoids protect astrocytes from ceramide-induced apoptosis
through the phosphatidylinositol 3-kinase/protein kinase B path-
way. J Biol Chem 2002; 277: 36527-33.
2298 Current Pharmaceutical Design, 2008, Vol. 14, No. 23
[75] Singh I, Pahan K, Khan M, Singh AK. Cytokine-mediated induc-
tion of ceramide production is redox-sensitive. Implications to
proinflammatory cytokine-mediated apoptosis in demyelinating
diseases. J Biol Chem 1998; 273: 20354-62.
[76] Blázquez C, Galve-Roperh I, Guzmán M. De novo-synthesized
ceramide signals apoptosis in astrocytes via extracellular signal-
regulated kinase. FASEB J 2000; 14: 2315-22.
[77] Carracedo A, Geelen MJ, Diez M, Hanada K, Guzmán M, Velasco
G. Ceramide sensitizes astrocytes to oxidative stress: protective
role of cannabinoids. Biochem J 2004; 380: 435-40.
[78] Carracedo A, Egia A, Guzmán M, Velasco G. p8 Upregulation
sensitizes astrocytes to oxidative stress. FEBS Lett 2006; 580:
1571-5.
[79] Back SA, Han BH, Luo NL, Chricton CA, Xanthoudakis S, Tam J,
et al. Selective vulnerability of late oligodendrocyte progenitors to
hypoxia-ischemia. J Neurosci 2002; 22: 455-63.
[80] Back SA, Gan X, Li Y, Rosenberg PA, Volpe JJ. Maturation-
dependent vulnerability of oligodendrocytes to oxidative stress-
induced death caused by glutathione depletion. J Neurosci 1998;
18: 6241-53.
[81] Zhou L, Trapp BD, Miller RH. Demyelination in the central nerv-
ous system mediated by an anti-oligodendrocyte antibody. J
Neurosci Res 1998; 54: 158-68.
[82] Aguado T, Palazuelos J, Monory K, Stella N, Cravatt B, Lutz B,
et al. The endocannabinoid system promotes astroglial differentia-
tion by acting on neural progenitor cells. J Neurosci 2006; 26:
1551-61.
[83] Parolaro D, Massi P. Cannabinoids as potential new therapy for the
treatment of gliomas. Expert Rev Neurother 2008; 8: 37-49.
[84] Guzmán M. Effects on cell viability. Handb Exp Pharmacol 2005;
168: 627-42.
[85] Pacher P, Bátkai S, Kunos G. The endocannabinoid system as an
emerging target of pharmacotherapy. Pharmacol Rev 2006; 58:
389-462.
[86] Petersen G, Moesgaard B, Schmid PC, Schmid HH, Broholm H,
Kosteljanetz M, et al. Endocannabinoid metabolism in human
glioblastomas and meningiomas compared to human non-tumour
brain tissue. J Neurochem 2005; 93: 299-309.
[87] Maccarrone M, Lorenzon T, Bari M, Melino G, Finazzi-Agro A.
Anandamide induces apoptosis in human cells via vanilloid recep-
tors. Evidence for a protective role of cannabinoid receptors. J Biol
Chem 2000; 275: 31938-45.
[88] Jacobsson SO, Wallin T, Fowler CJ. Inhibition of rat C6 glioma
cell proliferation by endogenous and synthetic cannabinoids. Rela-
tive involvement of cannabinoid and vanilloid receptors. J
Pharmacol Exp Ther 2001; 299: 951-9.
Massi et al.
[89] Maccarrone M, Pauselli R, Di Rienzo M, Finazzi-Agrò A. Binding,
degradation and apoptotic activity of stearoylethanolamide in rat
C6 glioma cells. Biochem J 2002; 366: 137-44.
[90] Contassot E, Wilmotte R, Tenan M, Belkouch MC, Schnüriger V,
de Tribolet N, et al. Arachidonylethanolamide induces apoptosis of
human glioma cells through vanilloid receptor-1. J Neuropathol
Exp Neurol 2004; 63: 956-63.
[91] Massi P, Vaccani A, Ceruti S, Colombo A, Abbracchio MP, Paro-
laro D. Antitumor effects of cannabidiol, a nonpsychoactive canna-
binoid, on human glioma cell lines. J Pharmacol Exp Ther 2004;
308: 838-45.
[92] Massi P, Vaccani A, Bianchessi S, Costa B, Macchi P, Parolaro D.
The non-psychoactive cannabidiol triggers caspase activation and
oxidative stress in human glioma cells. Cell Mol Life Sci 2006; 63:
2057-66.
[93] Manev H, Uz T, Sugaya K, Qu T. Putative role of neuronal 5-
lipoxygenase in an aging brain. FASEB J 2000; 14: 1464-9.
[94] Funk CD. Prostaglandins and leukotrienes: advances in eicosanoid
biology. Science 2001; 294: 1871-5.
[95] Sánchez C, Galve-Roperh I, Canova C, Brachet P, Guzmán M.
Delta9-tetrahydrocannabinol induces apoptosis in C6 glioma cells.
FEBS Lett 1998; 436: 6-10.
[96] Galve-Roperh I, Sánchez C, Cortés ML, del Pulgar TG, Izquierdo
M, Guzmán M. Anti-tumoral action of cannabinoids: involvement
of sustained ceramide accumulation and extracellular signal-
regulated kinase activation. Nat Med 2000; 6: 313-9.
[97] Herdener M, Heigold S, Saran M, Bauer G. Target cell-derived
superoxide anions cause efficiency and selectivity of intercellular
induction of apoptosis. Free Radic Biol Med 2000; 29: 1260-71.
[98] Schimmel M, Bauer G. Proapoptotic and redox state-related signal-
ing of reactive oxygen species generated by transformed fibro-
blasts. Oncogene 2002; 21: 5886-96.
[99] Duntsch C, Divi MK, Jones T, Zhou Q, Krishnamurthy M, Boehm
P, et al. Safety and efficacy of a novel cannabinoid chemotherapeu-
tic, KM-233, for the treatment of high-grade glioma. J Neurooncol
2006; 77: 143-52.
[100] Fletcher EL, Phipps JA, Ward MM, Puthussery T, Wilkinson-
Berka JL. Neuronal and glial cell abnormality as predictors of pro-
gression of diabetic retinopathy. Curr Pharm Des 2007; 13(26):
2699-712.
[101] Sathornsumetee S, Rich JN. Antiangiogenic therapy in malignant
glioma: promise and challenge. Curr Pharm Des 2007; 13(35):
3545-58.
[102] Walmsley AR, Mir AK. Targeting the Nogo-A signalling pathway
to promote recovery following acute CNS injury. Curr Pharm Des
2007; 13(24): 2470-84.