Chemico-Biological Interactions 312 (2019) 108819

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Chemico-Biological Interactions
journal homepage: www.elsevier.com/locate/chembioint

Cannabidiol improves metabolic dysfunction in middle-aged diabetic rats T

submitted to a chronic cerebral hypoperfusion
Maria Rosa Trentin Zorzenona,∗, Amanda Nunes Santiagob, Marco Aurélio Morib, Silvano Piovanc,
Cler Antônia Jansena, Maria Eduarda Perina Padilhaa, Simone Rocha Ciottaa,
Paulo Cezar de Freitas Mathiasc, Francisco Silveira Guimarãesd, Rubia Maria Weffort de Oliveirab,
Paula Gimenez Milania, Cecília Edna Mareze-Costae
a
Biochemistry Department, State University of Maringa, Av. Colombo 5790 CEP, 87020-900, Maringa, Parana, Brazil
b
Department of Pharmacology and Therapeutics, State University of Maringa, Av. Colombo 5790 CEP, 87020-900, Maringa, Parana, Brazil
c
Department of Biotechnology, Genetics and Cell Biology, State University of Maringa, Av. Colombo 5790 CEP, 87020-900, Maringa, Parana, Brazil
d
Centre for Interdisciplinary Research on Applied Neurosciences (NPNA), Department of Pharmacology, Medical School of Ribeirão Preto, University of São Paulo, Av.
Bandeirantes 3900 CEP, 14049-900, Ribeirão Preto, São Paulo, Brazil
e
Department of Physiological Sciences, State University of Maringá, Av. Colombo 5790 CEP, 87020-900, Maringa, Parana, Brazil

ARTICLE INFO ABSTRACT

Keywords: Cannabidiol (CBD), a compound obtained from Cannabis sativa, has wide range of therapeutic properties, in-
Cannabis sativa cluding mitigation of diabetes and neurodegeneration. Cerebral ischemia and consequent learning disabilities
Diabetes mellitus are aggravated in elderly diabetic subjects. However, there are no studies showing the effect of CBD treatment in
Cerebral ischemia elderly diabetes patients suffering cerebral ischemia. The present work tested the hypothesis that CBD treatment
Insulin
improves metabolic dysfunctions in middle-aged diabetic rats submitted to chronic cerebral hypoperfusion. In
Fructosamine
Middle-aged rats
this work, 350-day-old male Wistar streptozotocin-induced diabetic rats were used. To induce cerebral ischemia
was used a chronic cerebral hypoperfusion (CCH), surgically, via the four-vessel occlusion/internal carotid ar-
tery (4-VO/ICA). Four diabetic groups were established: Non-CCH Treated Diabetic (DNT), CCH Treated Diabetic
(DCT), Non-CCH Vehicle Diabetic (DNV), and CCH Vehicle Diabetic (DCV). Vehicle groups were not treated with
CBD. The animals were treated during 30 days with 10 mg CBD/Kg bw/day. After treatment, the animals were
euthanized, and blood levels of glucose, insulin, total cholesterol, high density lipoprotein (HDL), low density
lipoprotein (LDL), triglycerides, fructosamine, aspartate aminotransferase (AST) and alanine aminotransferase
(ALT) were evaluated. DCT group presented reduction of hyperglycemia and an increase of insulinemia. Also
was observed lower fructosamine, LDL, HDL, triglycerides and total cholesterol levels. AST and ALT con-
centration were reduced in CBD treated groups. CBD may be used as therapeutic tool to protect metabolism
against injuries from diabetes aggravated by cerebral ischemia.

1. Introduction
Cannabis sativa is a plant of the Cannabaceae family that has long been
cultivated for medicinal, textile; illicit and food purposes; therefore, it has
been a focus of attention of the scientific community and is consequently
one of the most studied plants worldwide [1]. Recently, a growing interest
has emerged in cannabis, resulting in increases in publications, reviews, and
clinical trials over time, largely due to the law acceptance of cannabis in
many countries [2] that are using cannabis-based products as pharmaceu-
ticals and even as food supplements [3].
Cannabis sativa leaves contain phytocannabinoids, which have
emerged as a new class of drugs with potential effects on a range of
neurodegenerative and psychiatric disorders [4]. The major phyto-
cannabinoids are tetrahydrocannabinol (THC) and cannabidiol (CBD);

Corresponding author.
∗

E-mail addresses: mariarosazorzenon@hotmail.com (M.R.T. Zorzenon), amandan.santiago@gmail.com (A.N. Santiago),

marcoaureliomori@gmail.com (M.A. Mori), silvanopionan23@gmail.com (S. Piovan), clerjansen@gmail.com (C.A. Jansen),
eduardapp@hotmail.com (M.E. Perina Padilha), simone.ciotta@hotmail.com (S.R. Ciotta), pmathias@uem.br (P. Cezar de Freitas Mathias),
fsguimar@fmrp.usp.br (F.S. Guimarães), rmmwoliveira@uem.br (R.M. Weffort de Oliveira), pgmfernandes2@uem.br (P.G. Milani),
cemcosta@uem.br (C.E. Mareze-Costa).

https://doi.org/10.1016/j.cbi.2019.108819
Received 25 April 2019; Received in revised form 21 August 2019; Accepted 6 September 2019
Available online 06 September 2019
0009-2797/ © 2019 Elsevier B.V. All rights reserved.
M.R.T. Zorzenon, et al.
Abbreviations
4-VO/ICA Four-vessel occlusion/internal carotid artery
ALT Alanine aminotransferase
AST Aspartate aminotransferase
CBD Cannabidiol
CCH Chronic cerebral hypoperfusion
DCT Diabetic group, CCH, treated with CBD
DCV Diabetic group, CCH, vehicle
however, THC has psychotropic actions and adverse side effects such as
hallucinations [5].
CBD is non-psychoactive and is associated with various beneficial
medicinal and therapeutic properties such as analgesic, antibacterial,
antidiabetic, antiemetic, antiepileptic, anti-inflammatory, anti-
proliferative, antipsychotic, and antispasmodic effects [6,7]. One of the
main goals of CBD studies is to examine its potential application for the
treatment of various neuropsychiatric conditions via its neuroprotective
effects on chronic cerebral hypoperfusion (CCH)-related neurodegen-
erative conditions, including diseases such as Parkinson's disease, epi-
lepsy, and dementias, such as Alzheimer's disease [8].
Aging and vascular comorbidities, such as hypertension and dia-
betes, significantly influence a wide variety of brain disorders [9]. A
meta-analysis of 28 studies showed that people with diabetes mellitus
(DM) had a 73% increased risk of developing dementia compared to the
general population [10].
Hyperglycemia, which is a characteristic of DM, usually causes
vascular lesions, which may aggravate the neurodegenerative state due
to microvascular changes, oxidative stress, chronic inflammation, and
accumulation of glycation products, which are also factors that con-
tribute to the cognitive effects of dementia [11,12].
Preexisting conditions that can aggravate the outcomes of CCH have
been examined. For example, hypertension interacted synergistically
with age to deteriorate the capacity of the brain to adaptively respond
to CCH [13]. In the same line, diabetes increases the mortality rate,
aggravates neurodegeneration and inflammation in the hippocampus
and white matter, and impairs cognitive performance in middle-aged
rats subjected to CCH [14]. Importantly, preclinical investigations of
the physiopathology of age-related pathologies as CCH, from combining
certain comorbidities as diabetes, offer insights to the development of
therapeutic strategies.
Studies show that CBD administration has great therapeutic poten-
tial for the treatment of streptozotocin (STZ)-induced DM, acting
mainly on oxidative stress, inflammation, and cell death [15,16]. Fur-
thermore, CBD has shown effects on biochemical parameters, such as
blood glucose, total cholesterol, high density lipoprotein (HDL) cho-
lesterol, and triglycerides, which are altered in diabetic patients
[5,17,18].
Santiago et al. [14] evaluated CBD treatment middle-aged diabetic
rats with cerebral ischemia using a four-vessel/internal carotid artery
occlusion (4-VO/ICA) model of CCH and demonstrated that CBD
treatment, in addition to reducing brain injury caused by CCH, also
reduced blood glucose levels during the treatment period. However,
little is known about the effects of CBD in elderly animals with these
pathologies and assessing these effects is of extreme importance be-
cause both pathologies mainly affect the elderly [19]. In addition,
evaluating possible peripheral and metabolic effects of treatment with
this compound is necessary to better understand its properties and ef-
fects in the prevention, treatment, and care of metabolic diseases such
as diabetes.
Thus, the objective of this study was to evaluate the effects of CBD
treatment on blood glucose and biochemical parameters related to STZ-
induced diabetes in middle-aged rats subjected to chronic cerebral
hypoperfusion.
2
Chemico-Biological Interactions 312 (2019) 108819
DM Diabetes mellitus
DNT Diabetic group, non-CCH, treated with CBD
DNV Diabetic group, non-CCH, not treated with CBD
HDL High density lipoprotein
ICA Internal carotid artery
LDL Low-density lipoprotein
ND Non-diabetic group, non-CCH, not treated with CBD
STZ Streptozotocin
VA Vertebral artery; VO, Vessel occlusion
2. Materials and methods
2.1. Animals
Male middle-aged wistar rats (350–380-day-old) [20–22] from the main
animal house of the State University of Maringa (UEM) were used. The
experimental protocols used in this study were approved by the UEM
Commission for Animal Experimentation and Use (protocol # 028/2014).
The animals were kept in the animal house of Cerebral Ischemia and
Neuroprotection Laboratory from UEM under the following conditions:
23 °C, 12 h light/12 h dark photoperiod, and ad libitum access to water and
balanced feed (Nuvilab brand - Colombo - PR). The animals were housed in
collective cages (46 × 24 × 20 cm) with 5 animals per cage.
2.2. Induction of diabetes and weekly biochemical parameter measurement
The STZ (2-deoxy-2-(3-methyl-3-nitrosoureido)-D-glucopyranose)-
induced DM model was used to establish the diabetic animal group.
These animals were fasted overnight for 12 h, given a single in-
travenous administration of STZ of 35 mg/kg body weight (bw) (Sigma,
St. Louis, MO, USA) dissolved in citrate buffer (0.10 mM; pH 4.5), and
then fasted for four more hours. Control animals received a saline in-
jection. STZ-treated animals received glucose (5%) after 24 h to reduce
mortality due to hyperglycemic shock [23]. Two days after STZ ad-
ministration, the blood glucose levels of the rats were measured using
an Optium mini® blood glucose monitoring system (Abbot Diabetes Care,
Inc., USA). Animals with fasting blood glucose levels that exceeded
250 mg/dL were considered diabetic [24].
2.3. Chronic cerebral hypoperfusion (CCH)
After 30 days of diabetes induction, the animals underwent CCH.
For CCH surgery, the 4-VO/ICA model was used and cerebral ischemia
was confirmed by retrograde memory loss as described by Santiago
et al. [25]. Control animals (non-CCH) underwent the same surgical
procedures but were not subjected to vessel occlusion.
2.4. Experimental design and CBD treatment
Five experimental groups were established: 1) ND = non-diabetic
group, no CCH, not treated with CBD; 2) DNV = diabetic group, no
CCH, not treated with CBD; 3) DNT = diabetic group, no CCH, treated
with CBD; 4) DCV = diabetic group, CCH, not treated with CBD; 5)
DCT = diabetic group, CCH, treated with CBD.
CBD (approximately 99% of purity provided by THC Pharma, Frankfurt,
Germany) was diluted in 1% Tween 80 and sterile isotonic saline (vehicle).
Treated animals received 10 mg/kg bw/day CBD intraperitoneally 30 min
prior to CCH surgery and during the 30-day period after surgery. The CBD
dose used was based on a previous study [8].
2.5. Blood glucose and biochemical parameters
2.5.1. Pre-prandial and post-prandial glycemic
Four blood samples were collected via caudal puncture to generate
M.R.T. Zorzenon, et al. Chemico-Biological Interactions 312 (2019) 108819

the blood glucose curve. The first sample was collected when the ani-
mals were undergoing 12-h fasting, this was considered pre-prandial
glycemic. Immediately after, the animals were fed ad libitum, and after
2 h, additional samples were collected and considered post-prandial
glycemic (after being fed). Glycemia was determined using an Optium
mini® blood glucose monitoring system (Abbot Diabetes Care, Inc USA).

2.5.2. Euthanasia and organ and tissues collection

Animals were anesthetized by inhalation of an oxygen and halothane
mixture (Tanohalo®, Cristália, Itapira-SP, Brazil). They were then eu-
thanized (guillotine) and underwent a median laparotomy to collect blood
and tissue samples. Blood was transferred to heparinized plastic tubes and
centrifuged (4 °C, 15 min, 2000 rpm). The obtained plasma was stored in a
freezer at −20 °C and subsequently used for biochemical measurements.
Brown and periepididymal fat, testes, seminal vesicles, kidneys, the liver,
and the spleen were weighed and stored in a −20 °C freezer. Fig. 2. Effect of CBD on fasting insulinemia in middle-aged, diabetic and
chronic cerebral hypoperfusion rats. DNV: Vehicle Non-CCH Diabetic
2.5.3. Biochemical measurements (n = 6); DNT: Treated Non-CCH Diabetic (n = 8); DCV: Vehicle CCH Diabetic
(n = 7); DCT: Treated CCH Diabetic (n = 9). Values represent the
Fructosamine, triglycerides, total cholesterol, HDL cholesterol, as-
mean ± S.P.M. **p < 0.01 between DNV and DNT, *** < 0.001 between
partate aminotransferase (AST) and alanine aminotransferase (ALT)
DCV e DCT, **** < 0.0001 between DCT e DNV.
levels were determined using Gold Analisa kits (Gold Analisa
Diagnostica Ltda, Belo Horizonte, Minas Gerais, Brazil). The plasma
insulin concentration was determined by a radioimmunoassay method,
according to Piovan et al. [26].

2.6. Statistical analysis

The results, which are presented as the mean ± standard error of the
mean (SEM), were subjected to analysis of variance (ANOVA) and Tukey's
test (p < 0.05). GraphPad Prism version 7.0 (Windows GraphPad Prism
Software, San Diego, CA, USA) was used for the statistical analysis.

3. Results

3.1. Blood glucose and insulin

Fig. 1 shows pre-prandial and post-prandial glycemia for the ex-

perimental groups. The DCT group showed significantly lower blood
glucose levels pre-prandial and post-prandial, 14.5% (p < 0.05) and
14.1% (p < 0.001), respectively, compared to DCV group.
Fasting insulinemia is showed in Fig. 2. Diabetic rats that received CBD
treatment had significantly greater plasma insulin levels than those that
received the vehicle alone; 126.1% (DNT vs DNV; p < 0.01) and 74.3%
(DCT vs DCV; p < 0.001). Fig. 2 also shows that the DCT group presented a
substantial rise to insulinemia in comparison to the DNV group
(p < 0.0001), i.e., diabetic animals treated with CBD, subjected to cerebral
vessels occlusion had higher plasma insulin concentrations.
3.2. Biochemical analyses
Fasting glucose levels of diabetic animals did not present any
Fig. 3. Effect of CBD on fasting fructosamine level in middle-aged, diabetic
and chronic cerebral hypoperfusion rats. DNV: Vehicle Non-CCH Diabetic
(n = 7); DNT: Treated Non-CCH Diabetic (n = 9); DCV: Vehicle CCH Diabetic
(n = 7); DCT: Treated CCH Diabetic (n = 9). Values represent the
mean ± S.P.M. **p < 0.005 between DCV and DCT; ***p < 0.001 between
DNV and DCT.
difference, regarding cerebral ischemia (DNV 542.9 and DCV
471.2 mg/dL), neither to diabetic animals treated with CBD (DNT 474.7
and DCT 503.3 mg/dL).
Fructosamine blood levels were decreased 26% (p < 0.005) in DCT
compared to DCV and 31.5% (p < 0.001) compared to DNV, as shown
in Fig. 3.

Fig. 1. Effect of CBD on pre-prandial (A)

and post-prandial glycemia (B) of
middle-aged, diabetic and chronic cere-
bral hypoperfusion rats. DNV: Vehicle
Non-CCH Diabetic (n = 6); DNT: Treated
Non-CCH Diabetic (n = 8); DCV: Vehicle
CCH Diabetic (n = 7); DCT: Treated CCH
Diabetic (n = 8). Values represent the
mean ± S.P.M. *p < 0.05 between DCV
and DCT; **p < 0.005 between DNV and
DCT; ***p < 0.001 between DNV and
DCT; DCV and DCT; ****p < 0.0001 be-
tween DNT and DCT.

3
M.R.T. Zorzenon, et al.
Regarding lipid parameters (Table 1), DCT group presented a sig-
nificant reduction, 22.3% (p < 0.05) in total cholesterol compared to
the DCV group. Total triglycerides also were reduced by half
(p < 0.05) in DCT compared to DCV rats. However, diabetic non-CCH
rats (DNT) had a three-fold increase in triglycerides when treated with
CBD. Non expectedly, HDL concentration in the DCT group was re-
duced, 39.1% (p < 0.05) compared to DCV group.
Table 1
Biochemical parameters of CBD in diabetic middle-aged rats submitted to chronic cerebral hypoperfusion.
Analyzes DNV
#
Total Cholesterol (mg/dL) 74.80 ± 6.66
Cholesterol HDL (mg/dL) 41.70 ± 2.69
Triglycerides (mg/dL) 80.00 ± 16.06&
ALT (U/L) 220.7 ± 19.01
AST (U/L) 139.3 ± 14.51
DNV: Vehicle Non-CCH Diabetic (n = 7); DNT: Treated Non-CCH Diabetic (n = 9); DCV: Vehicle CCH Diabetic (n = 6); DCT: Treated CCH Diabetic (n = 12). Values
represent the mean ± S.P.M. #p < 0.05 differs from DCV; +p < 0.05 differs from DCV and DNT; &p < 0.05 differs from DNT; ##p < 0.05 differs from DCV
and DNV.
Both groups treated with CBD (DCT and DNT) had AST reduction,
61.5% and 61% (p < 0.05) compared with its respective controls, DCV
and DNV rats (Table 1). Although ALT presented a reduction of 35%
(p < 0.05) from DNT compared to DNV, there was no difference be-
tween CCH animals.
Regarding organ and tissue weight, CBD treatment did not sig-
nificantly alter any of the results obtained (data not shown).
4. Discussion
Middle-aged diabetic animals with cerebral ischemia treated with
CBD showed a reduction in hyperglycemia. In addition, these animals
showed an increase in insulin secretion and a reduction in diabetes
complications, such as advanced glycation end-products, dyslipidemia
and hepatic complications.
The decrease in hyperglycemia was possibly caused by increased
insulin secretion. Hyperglycemia in elderly subjects is directly related
to changes in glucose tolerance that are associated with defects in the
action and/or secretion of insulin, clinical signals that characterising
type 2 DM [27].
High dose of STZ (60–100 mg/kg body weight) in adults rats, has
been shown to emulate a induction to type 1 DM, leading to total de-
struction of pancreatic β-cells and consequently suspending out insulin
production; whereas the low dose of STZ (30–50 mg/kg body weight)
induces a mild impairment of insulin secretion, similar to the late stages
of type 2 DM. Although humans and rats with type 2 DM present per-
ipheral tissue insulin resistance, β-cells can secret insulin which means
that it can produce, accumulate and release insulin barely controlled by
many factors such as oscillation of blood glucose concentration, hor-
monal and neuronal signals, leading to β-cells progressive malfunction
[28].
The results suggest that CBD caused insulin secretion stimulation in
diabetic animals with cerebral ischemia and consequently reduced
hyperglycemia, similar to the result obtained by Ruz-Maldonado et al.
[29], whose demonstrated an increased insulin secretion by pancreatic
islets in humans and rats treated with a synthetic CBD analog. The
possibility of CBD to induce β-cells proliferation cannot be eliminated,
which means insulin secretion induced by CBD has a relation to the
number of functional β-cells [30]. A histological study of isolated islets
showed that CBD reduced the degree of severity of female mice with
insulitis [31], indicating that CBD could have a protector effect against
progressive dysfunctions on β-cells. In another study, treatment with
CBD showed a protective effect, increasing delay of type 2 DM onset by
pancreas inflammation. The authors concluded that CBD acted as an
anti-inflammatory agent [5].
Chemico-Biological Interactions 312 (2019) 108819
Some studies have shown that CBD effects are transduced by CBD
receptors, mainly CB1, CB2, GPR55 and PPARγ. CB2 receptors are
mostly present in the peripheral organs; whereas, CB1 is mainly present
in the central nervous system and, to a lesser extent, in other tissues,
including pancreas [32].
In particular, CB1 receptors are expressed in pancreatic β-cells [33].
Blockage of CB1 receptor caused by CBD, increases insulin secretion [34].
DNT DCV DCT
99.22 ± 5.47 108.8 ± 5.23 84.5 ± 2.72#
59.50 ± 3.32 53.80 ± 3.60 32.80 ± 2.39+
252.80 ± 32.7 136.7 ± 18.52 69.20 ± 9.18+
143.4 ± 5.79## 215.1 ± 15.33 186.8 ± 14.09
54.10 ± 5.51## 166.9 ± 19.77 64.40 ± 5.51##
Another receptor is GPR55, its blocking exhibited insulinotropic
properties in clonal β-cells, mice isolated islets and in vivo in mice [35].
However, Liu et al. [36] investigating the mechanism of action of CBD
in islets isolated from a transgenic mouse model with homozygous
deletion of the GPR55 gene, showed CBD-induced insulin secretion,
suggesting that CBD transduces its effects via a signaling pathway dif-
ferent from GPR55 interaction. On the other hand, CBD acts as a
complete agonist of PPARγ, which is a family of nuclear hormone re-
ceptors and is predominantly expressed in adipose tissue, having a
major role in promoting adipocyte differentiation, fatty acid storage,
insulin sensitivity and glucose metabolism, whereas PPARγ is also
found throughout the brain mediating the inflammatory cascade [37].
Therefore, CBD can mediate neuroprotective effects by activating pro-
inflammatory pathways and possibly mediate effects on insulin sensi-
tivity in peripheral tissue and glucose metabolism.
There is a relationship between dementia and hyperinsulinemia,
which is expressed as compensatory mechanism, because of the insulin
resistance in the peripheral tissues [38,39]. These implications are
presented in type 2 DM. Although the diabetic rats used in the present
study do not present insulin resistance, they are hyperglycemic and
treatment with CBD reduces it.
Brain injuries exacerbate central insulin resistance, and prolonged
exposure to hyperglycemia and hyperinsulinemia can lead to dete-
rioration of neuronal structure and function, resulting in poor cognitive
performance [40]. Considering that insulin mostly induces glucose
uptake in insulin-dependent organs and tissues, it has also, neurotropic
properties. It has been observed insulin receptors in brain areas asso-
ciated to learning and memory activity [38].
Strikingly, it has been shown that insulin improved cognitive ac-
tivity in patients whose present memory debilities. Santiago et al. [14]
demonstrated that CBD administration in diabetic animals subjected to
CCH resulted in neuroprotective effects by preventing memory deficit,
reducing neurodegeneration and reduction hyperglycemia. These re-
sults suggest that CBD may effectively reduce the risk of dementia as-
sociated to diabetes. Whether or not the improvements caused by CBD
administration are effects caused by itself or by increased blood insulin
concentration can be a matter to future studies.
Diabetic animals with cerebral ischemia and treated with CBD
presented a reduction in fructosamine. This suggests that CBD treat-
ment could reduce DM-associated complications because advanced
glycation end-products (AGEs), inferred by fructosamine blood levels,
are associated with cognitive impairment and moderate declines in
motor speed and psychomotor efficiency [38,39]. In addition, diabetic
individuals present high levels of AGEs. Indeed, insulin therapy reg-
ulates glucose homeostasis and also decreases of AGE levels [40].
4
M.R.T. Zorzenon, et al.
The increase in insulin blood levels caused by CBD may be indirectly
associated with the reduction of AGEs, by the control of glucose
homeostasis. Rajesh et al. [16] showed that STZ-induced diabetic ani-
mals treated with different CBD concentrations (1, 10, and 20 mg/kg)
did not affect blood glucose, similar to the results found in the present
study, which show that diabetic animals without cerebral ischemia did
not present a substantial reduction of hyperglycemia. CBD adminis-
tration also had no effect on blood glucose in type 2 DM patients [41].
However, Lehmann et al. [5], who treated mice with CBD prior to the
induction of type 1 DM via inflammation of the pancreas, showed that
CBD had anti-inflammatory activity, delaying the appearance of clinical
signs of DM, including hyperglycemia.
McKillop et al. [35] showed a 19% decrease in triglycerides and a
17% decrease in total cholesterol, as well as a 19% increase of HDL
cholesterol in STZ-induced diabetic animals treated with a synthetic
CBD analog. Similar results were observed in the current study. It has
been shown that dyslipidemia observed in diabetic rats improved by
insulin therapy [40]. However, it was shown that CBD can act directly
to normalize lipid profile in diabetic subjects [41]. Again, we do not
present current evidence to suggest that CBD acts directly or indirectly
in these diabetic middle-aged animals submitted to CCH, however, we
can speculate that, at least in part, CBD increasing insulin levels could
be responsible by metabolic improvement.
Enzymes AST and ALT are used as markers of liver injury. The
treatment with CBD presented reduction in these enzymes, which have
increased levels in diabetic subjects according to the degree of hepatic
injury [42]. These possible protective effects were also found by Co-
melli et al. [43], who showed that 7 days of treatment with CBD-rich
cannabis leaf extract significantly decreased liver damage, providing
protection against oxidative stress due to the antioxidant activity of
CBD. Other studies in vitro and clinical have also shown a hepatopro-
tective effect of CBD [44,45]. The beneficial effects of CBD treatment on
diabetes appear to be mainly mediated by the anti-inflammatory and
antioxidant effects of CBD [46]. Yang et al. [47] concluded that rats
with cerebral ischemia through middle cerebral artery occlusion had
elevated ALT and AST activities at serum levels compared to un-
operated animals, indicating that liver function was impaired. No stu-
dies were found to quantify AST and ALT in animals submitted to
cerebral ischemia by chronic cerebral hypoperfusion (CCH). Thus, our
work is pioneering in assessing biomarkers of liver injury in correlated
pathologies such as diabetes and cerebral ischemia, and further in-
dicates that CBD treatment reduces these metabolic disorders. How-
ever, our study shows that ALT levels in rats who were diabetic, suf-
fered cerebral ischemia and were treated with CBD declined, although
the different is not statistically significant. Interestingly, AST levels
were reduced in diabetics rats with cerebral ischemia and treated with
CBD, and this suggests that CBD directly or indirectly can mitigate
hepatic functions.
Santiago et al. [14] showed that diabetes combined with CCH af-
fected neurodegeneration and memory more severely in middle-aged
rats than in young rats, suggesting that diabetes interacts synergistically
with age in decreasing brain responses. In addition, other biological
dysfunctions are caused by the association of age and diabetes, in-
cluding changes in lipid metabolism, cellular respiration, and reactive
oxygen species [9].
Hyperglycemia is usually accompanied by other comorbidities, in-
cluding insulin resistance, obesity, and inflammation; therefore, it is
very difficult to determine the specific effect of hyperglycemia itself on
brain function [48]. However, compounds with high antioxidant ac-
tivity are known to be beneficial for brain function and diabetes [49].
Among these, as shown in the current study, CBD improved metabolic
dysfunctions presented in diabetic rats with severe cerebral ischemia.
5. Conclusion
Treatment with CBD caused a reduction in pre and post-prandial
5
Chemico-Biological Interactions 312 (2019) 108819
glycemia of middle-aged diabetic rats submitted to CCH. CBD was also
able to increase insulin levels and decrease AGEs levels. However, the
effects of cannabinoid on dyslipidemia have been controversial and
studies are needed to evaluate the mechanism of action of CBD on lipid
profile. These initial results suggest that CBD may act as an important
pharmacological agent to prevent metabolic dysfunction in diabetic
patients with cerebral ischemia. In addition, it can be suggested that
Cannabidiol could be used as a drug to mitigate neuronal degeneration
caused by diabetes associated to vascular cerebral accidents, regulating
the metabolism. In this study, the effects of CBD on associated pathol-
ogies, diabetes and cerebral ischemia were evaluated. However, studies
to investigate their effects individually in these pathologies are needed
to better elucidate the mechanism of this cannabinoid.
Funding
This study was supported by Brazilian National Council for
Scientific and Technological Development (Conselho Nacional de
Desenvolvimento Científico e Tecnológico - CNPq - National Institute of
Science and Translational Medicine, Grant numbers: 465458/2014-9)
and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior -
Brasil (CAPES) - Finance Code 001.
Conflicts of interest
The authors have declared that there is no conflict of interest.
Declaration of interests
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
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