Research Article
Hydro-alcoholic Extract of Commiphora mukul Gum Resin May Improve Cognitive Impairments in Diabetic Rats
Iraj Salehi 1, Zahra Taheraslani 2, Shirin Moradkhani 4 *
1 Neurophysiology Research Center, Hamadan University of Medical Sciences, Hamadan, IR Iran
2 Department of Physiology, School of Basic Sciences, Hamadan Branch, Islamic Azad University, Hamadan, IR Iran
3
Department of Pharmacognosy and Pharmaceutical Biotechnology, School of
Pharmacy, Hamadan University of Medical Sciences, Hamadan, IR Iran
*Corresponding
author: Shirin Moradkhani, Department of Pharmacognosy and
Pharmaceutical Biotechnology, School of Pharmacy, Hamadan University of
Medical Sciences, Hamadan, IR Iran. Tel: +98-8138381593, Fax:
+98-8138380031, Email: shirin.moradkhani@yahoo.com
Abstract
Background: Diabetes
causes cognitive impairment. Medicinal plants due to different
mechanisms, such as antioxidant activities may improve diabetes and
relieve its symptoms. Commiphora mukul (Burseraceae) has a significant antioxidant activity.
Objectives: This study aimed to examine the effect of hydro- alcoholic extract of C. mukul on passive-avoidance learning and memory in streptozotocin (STZ) induced diabetic male rats.
Materials and Methods: Thirty-two adult male Wistar rats were randomly allocated to four groups: normal, diabetic, normal + extract of C. mukul and diabetic + extract of C. mukul
groups with free access to regular rat diet. Diabetes was induced in
male rats by single interaperitoneal injection of 60 mg/kg STZ. After
the confirmation of diabetes, 300 mg/kg C. mukul extract was
orally administered to the extract-treated groups. Control groups
received normal saline at the same time. Passive-avoidance memory was
tested eight weeks after the STZ treatment, and blood glucose and body
weight were measured in all groups at the beginning and end of the
experiment.
Results: In the present study, diabetes decreased learning and memory. Although the administration of C. mukul
extract did not affect the step-through latency (STLa) and the number
of trials of the diabetic groups during the first acquisition trial, a
significant decrease was observed in STLr and also a significant
increase in time spent in the dark compartment (TDC) and number of
crossing (NOC) in the retention test (after 24 and 48 hours). Although
no significant difference was observed in body weight of diabetic +
extract of C. mukul (DE) and diabetic control (DC) groups, the plasma glucose of DE group was significantly lower in comparison to DC group.
Conclusions: Commiphora mukul
extract can improve passive-avoidance learning and memory impairments
in the STZ-induced diabetic rats. This improvement may be due to the
antioxidant, acetylcholinesterase inhibitory activity, anti-inflammatory
or power to reduce hyperglycemia and lowering cholesterol and
triglyceride properties of this extract. The result of this study is a
promising point for new therapeutic alternatives in alleviating
cognitive impairments caused by diabetes.
Keywords: Commiphora mukul; Passive Avoidance; Learning; Memory; Diabetes Mellitus
1. Background
Diabetes mellitus is a
metabolic disorder of endocrine, which characterized by chronic
hyperglycemia with disturbances of carbohydrate, fat and protein
metabolism because insulin production is inadequate, or because the
body's cells do not respond properly to insulin, or both. International
Diabetes Federation has estimated the incidence of diabetes for year
2030 is 350 million (1).
Therefore, public health concerns about poor academic
performance and memory impairment in children and adults with diabetes,
respectively. Recent evidences show that the central nervous system is
affected by diabetes, which is independent from atherosclerotic disease.
In humans, diabetes causes moderate cognitive dysfunctions and patients
are at a high risk of dementia and Alzheimer diseases. Cognitive
impairments also occur in diabetic rats, which are induced by
streptozotocin (STZ). This dysfunction can be as a result of structural
and functional deficits in hippocampus and cerebral cortex (2).
The pathogenesis of learning and memory impairments in diabetes
has multiple etiologies such as metabolic disorders, chronic
inflammation, vascular complications, and the dysfunction of scavenging
free radical systems. Up to now, there is no specific and effective
treatment for the management of cognitive impairments due to diabetes (3).
Nootropic agents (e.g. vitamins, fatty acids, antioxidants,
amino acids, minerals, and phytochemicals) refer to memory enhancers,
cognitive enhancers and neuroenhancers that can improve mental
functions(4).
According to World Health Organization estimation, 75% of the
world's population use traditional remedies and many synthetic drugs are
analogues of phytochemicals (5).
Commiphora mukul belongs to Burseraceae family of plant
kingdom. It has a resinous secretion known as guggul. Guggul is one of
the most valuable remedies in one of the traditional system of medicine
like the ayurvedic medicine. Its usage backs to over 2500 years. Guggul
has been used as antidiabetic, antihyperlipidemic, antiosteoarthritic,
anti-inflammatory, antispasmodic, sedative, antiseptic, astringent,
carminative, emmenagogue, expectorant, diaphoretic, thyroid stimulant,
antiobesity, diuretic, anthelminitic, and demulcent in traditional
medicine. It has no well-defined chemical composition due to its very
complex nature. Phytochemical investigation of guggul indicates the
presence of sugars (fructose, sucrose), amino acids, oil and several
steroids. However, only guggulsterone has been purified from the
ethyl-acetate extract of resin (6).
Guggulsterone is the active sterol of the plant, as an
antagonist of the bile acid farnesoid X receptor, demonstrating the
hypolipidemic effect of the resin. In the STZ-induced model of dementia,
guggulsterone has shown protective effects which is attributed to
cholesterol-lowering antioxidant and antiacetylcholine esterase
properties of this steroid. Hence, guggulsterone can be a potential
antidimentia drug (4, 7).
2. Objectives
Our literature survey indicated that there is no report on the effect of hydroalcoholic extract of C. mukul resin
on passive-avoidance learning and memory deficits in the STZ-induced
diabetic rats. Therefore, the purpose of the present study was to
investigate whether the administration of hydro-alcoholic extract of C. mukul resin has any protective effect on cognitive impairments in the STZ-induced diabetic rats.
3. Materials and Methods
3.1. Plant Extraction
Commiphora mukul gum resin was purchased from market in
Hamadan, Iran. Then 200 g of resin was macerated in ethanol 80% for 3
days. This process was repeated 3 times. The resulting extract was
filtered. The filtered extract was then concentrated to dryness in a
rotary evaporator under reduced pressure at a constant temperature of
40˚C. The resulting extract was stored in a refrigerator.
3.2. Animals
Thirty-two male Wistar rats (250 – 280 g) were purchased from
animal house, Hamadan University of Medical Sciences. The condition of
maintenance was the constant temperature (22 ± 0.5˚C) with 12-hour light
and 12-hour dark cycles. All animals had free access to a regular rat
diet. The experimental groups were chosen from different cages, randomly
with 8 animals per group.
3.3. Experimental Design
The animals were divided into two diabetic and two control groups
(n = 8 each). Diabetes was induced by a single intraperitoneal
injection of STZ (Sigma-Aldrich, Germany) (60 mg/kg). After three days,
fasting blood glucose levels were determined. Blood samples were
collected from the tail vein, and plasma glucose was measured using a
glucometer. Animals were considered diabetic if plasma glucose levels
exceeded 250 mg/dL. As soon as diabetes was confirmed, both diabetic and
normal groups received saline or 300 mg/kg of the extract (8)
by oral gavage for 60 days. At the end of the experiment, all rats were
weighed and blood was collected for plasma glucose measurement.
3.4. Passive Avoidance Learning
A light/dark shuttle-box, the apparatus for quantification of the
passive avoidance learning, had illuminated compartments of the light
and dark parts with a guillotine door between them. Five seconds after
placement of rat in the lighted part, the door was raised and habitually
it entered into the dark part. After the rat entrance, it was trapped
in dark part and a 1.2-mA constant current shock with 50 Hz frequency in
1.5 seconds was applied. The process of training was repeated until the
rat did not cross to dark part and remained in lighted part for 120
seconds. This process was repeated 24 and 48 hours after the acquisition
trial as a retention test. The retention performance was quantified by
the step-through latency during the retention trial (STLr), time spent
in the dark compartment (TDC) and number of crossing (NOC). At 8:00 –
11:00 hours the behavioral tests were performed and the ceiling score
was 600 seconds.
3.5. Plasma Glucose Level Measurement
At the end of the experiment, all rats were weighed and
decapitated under ketamine HCl (50 mg/kg, i.p.). Measurement of plasma
glucose levels was done using a glucometer.
3.6. Statistical Analysis
The data were analyzed by SPSS 21, shown as mean ± SEM and
compared by one-way ANOVA, post-hoc test LSD. The P Values of less than
0.05 were considered to be significant.
4. Results
4.1. Effects of Diabetes on the Passive-Avoidance Learning and Memory
One-way ANOVA indicated no significant difference in the STLa and
in the number of trials of the diabetic and control groups during the
first acquisition trial before the administration of the electrical
shock; (P > 0.05, Table 1).
During the retention test, after 24 and 48 hours the diabetic group had
a decreased STLr, and increased TDC and NOC compared to the control
group, respectively (P < 0.001), Table 1.
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Table 1.
Effect of Commiphora mukul Hydro-alcoholic Extract on Passive-Avoidance Learning and Memory in Rats a
|
4.2. Effects of Commiphora mukul Administration on Passive-Avoidance Learning and Memory in Nondiabetic Rats
There was no significant difference in the STLa and in the number
of trials of the control rats treated with extract and untreated
control rats during the first acquisition trial (before the
administration of the electrical shock; P > 0.05, Table 1).
Also, during the retention test, after 24 and 48 hours the control rats
treated with extract had no significant difference in STLr, TDC, NOC
compared to the untreated control rats (P > 0.05; Table 1).
4.3. Effects of Commiphora mukul Administration on Passive-Avoidance Learning and Memory in Diabetic Rats
There was no significant difference in the STLa and in the number
of trials of the extract-treated diabetic rats and untreated diabetic
rats during the first acquisition trial (before the administration of
the electrical shock; P > 0.05, Table 1).
During the retention test, after 24and 48 hours, the extract-treated
diabetic rats had an increased STLr and decreased TDC, NOC compared to
the untreated diabetic rats, respectively (P < 0.001, P < 0.05), Table 1.
4.4. Effects of Commiphora mukul Administration on Body Weight and Plasma Glucose
There was a significant increase in blood glucose and also a
significant decrease in body weight in diabetic rats as compared to the
normal rats. The oral administration of extract of C. mukul
significantly lowered the blood glucose in comparison with the untreated
group of diabetic rats; however, there was no significant increase in
body weight, as they were compared with the untreated group of diabetic
rats, (Table 2).
In Table 2,
the blood glucose levels and body weight of all experimental groups at
the beginning and at the end of the study are shown. Plasma glucose
levels between all groups at the onset of treatment were significantly
different. Eight weeks after the induction of diabetes, behavioral
assays were done and then plasma glucose levels and body weight were
measured. The body weight of the untreated (190.37 ± 6.50 g) and extract
-treated diabetic rats (208.12 ± 6.18 g) were lower than the control
rats (310.62 ± 6.97 g). Furthermore, there was no significant difference
in the body weight of the extract-treated (321.50 ± 5.72 g) and
untreated control (310.62 ± 6.97) animals. Regarding plasma glucose
levels, the untreated diabetic animals had significantly (P < 0.001)
elevated plasma glucose levels (517.62 ± 15.54 mg/dL) compared to the
control animals (112.87 ± 2.94 mg/dL). Oral administration of the C. mukul
extract to diabetic rats decreased the plasma glucose levels of the
treated groups (464.25 ± 17.36 mg/dL) compared to the untreated diabetic
group (517.62 ± 15.54 mg/dL; P < 0.05).
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Table 2.
Body Weight and Plasma Glucose Levels of Different Animal Groups at the Beginning and End of the Study a
b
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5. Discussion
According to the results of this study, the oral administration of hydro-alcoholic extract of C. mukul
gum resin improved passive-avoidance learning and memory of the control
rats and alleviated the negative influence of diabetes on learning and
memory. Although there was no significant effect in the STLa and in the
number of trials of the diabetic groups during the first acquisition
trial, a significant decrease in STLr and also a significant increase in
TDC, NOC in the retention test (after 24 and 48 h) showed promising
results of the present study.
Literature survey revealed that C. mukul had a profound effect on the antioxidant defense system; however, there was no report on the effect of C. mukul in
passive-avoidance learning and memory in diabetic rats. Diabetes is
associated with an increased production of reactive oxygen species
(ROS), enhanced oxidative stress and changes in the antioxidant capacity
(9, 10).
Oxidative stress is involved in the pathogenesis of many central
nervous system disorders (e.g. neurodegenerative diseases) or in the
physiological process of aging (11).
According to the literature, brain is very vulnerable to
oxidative stress due to its high-polyunsaturated fatty acids (PUFAs)
content, which are particularly susceptible to ROS damage (12, 13).
Apoptosis of nerve cells as a result of increased
Thiobarbituric acid reactive substances (TBARS) (lipid peroxidation
productions) in frontal cortex and hippocampus, induces the dysfunction
of learning and memory due to the structural and functional changes in
biological membranes induced by lipid peroxidation (14).
Impairment of learning and memory in the diabetic groups may be
related to the increased oxidative stress in diabetic animal's brain,
but C. mukul as an antioxidant may reduce the oxidative stress,
and lead to the better behavioral activity of the animals in
extract-treated diabetic groups. Clinical and experimental studies
suggest that hyperglycemia and/or insulin deficiency itself may be
responsible for impaired cognitive function in type1diabetes (15).
Moreover, antioxidants, antihyper-glycemics and insulin sensitizing agents can reduce cognitive dysfunction in diabetes (16).
The underlying mechanism of its action may be attributed to its
antioxidant, anti-inflammatory or acetylcholinesterase (AChE) inhibitory
properties. In vitro evaluation of C. mukul gum resin extract revealed that it has a profound effect on the antioxidant defense system (10) and can affect parameters of oxidative stress glutathione and malondialdehyde (GSH and MDA). Commiphora mukul,
with profound antioxidant potential and having the ability to trigger
cellular antioxidants, can be exploited for its use against a number of
disorders including cardiovascular diseases, inflammation, and cancer (17).
Commiphora mukul has been reported to contain flavonoids, terpenes, and phytosterols (18).
Flavonoids are potent antioxidants at very low concentrations. The
chemo-preventive properties of flavonoids are generally believed to
reflect their ability to scavenge the endogenus ROS.
By inhibiting or stimulating various signaling pathways, flavonoids at low concentration could affect cellular function (19, 20).
Acetylcholinesterase expression is pivotal during apoptosis. Indeed,
the AChE inhibitors partially protect cells from apoptosis. During the
type 1 diabetes mellitus development, an accumulation of AChE in the
pancreatic islets induces apoptotic βcells. Decrease in hyperglycemia
and incidence of diabetes, and restoration of plasma insulin levels and
plasma creatinine clearance are the consequences of treatment with AChE
inhibitors.
Induction and accumulation of AChE in pancreatic islets and the
protective effects of AChE inhibitors on the onset and development of
type 1 diabetes indicate a close relationship between AChE and type 1
diabetes (21).
AChE inhibitors enhance cholinergic function in the brain when loss or
decline in memory and cognitive impairment has occurred (22, 23). Commiphora mukul can increase whole brain AChE enzyme inhibitory activity (24). Polyphenols have shown AChE inhibitory effect (25) and considering that C. mukul contains flavonoids (18), another possible mechanism of C. mukul effect on passive avoidance learning can be involved in the AChE inhibitory potential of plant. Besides flavonoids, C. mukul is rich in guggulsterone. On the other hand, antioxidant and anti-inflammatory effects are relevant to each other (26).
Hence, guggulsterone , a phytosterol with potent anti-inflammatory
activity can be responsible for some of beneficial effects of C. mukul (27). Also, the extract affected plasma glucose.
The improvement of cognitive function observed in the diabetic animals in this study may be partly due to the ability of C. mukul
to attenuate hyperglycemia. In both human and animal models, guggul
lowered both serum low-density lipoprotein (LDL) cholesterol and
triglyceride levels, due to antagonistic effect of guggul on the bile
acid receptor farnesoid X-receptor. Biochemical and epidemiological
studies demonstrated a link between cholesterol, amyloid precursor
protein processing and Alzheimer’s disease. Cholesterol lowering drugs
decreased neuronal cholesterol levels, which in turn inhibit the beta
amyloid-forming amyloidogenic pathway, possibly by removing amyloid
precursor protein from cholesterol and sphingolipid-enriched membrane
micro domains. These results indicate that the administration of
cholesterol-lowering drugs is associated with decreased prevalence of
Alzheimer’s disease (7).
In conclusion, the findings of this study indicate that C. mukul
can improve cognitive impairment in diabetic rats. Cellular and animal
studies as well as clinical trials are required to support its role as
useful preparation in alternative medicine.
Acknowledgments
The authors are grateful to Neuroscience Research
Center, Hamadan University of Medical Sciences, Hamadan, Iran, for its
financial supports.
Footnotes
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