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49 International Journal of Natural Products Research 2011; 1 (4):49-54
ISSN: 2249-0353
Original Article Lipid lowering and hypoglycaemic potential of dried Dalbergia latifolia Roxb. bark extract in
Sprague-Dawley rats induced with high fat diets.
Mohammad Khalid*1, H.H. Siddiqui
1
1Faculty of Pharmacy Dasauli Kursi road, Integral University, Lucknow, India. 226026.
*Email: [email protected]
Received 14 October 2011; accepted 31 October 2011
Abstract
The present study was designed to investigate the hypolipidemic and hypoglycaemic activity of Dalbergia latifolia with regard to protection of LDL oxidation and disturbances of carbohydrates, fats and protein metabolism in-vivo study in rats made
hypercholesterolemic by continuous feeding of high fat diet. It contains dalbinol a new 12a-hydroxyrotenoid, sisafolin
coumarin from seeds, β- sitosterol, also contain dalbergichromene, lupeol, latifolin, and dalbergin from bark. Administering the
dried Dalbergia latifolia bark extracts (DL-extract) at doses of 100, 200 and 400 mg/kg daily together with continuous high fat
diet feeding to hypercholesterolemic rats for 8 weeks significantly decreased serum cholesterol level by 139.84±1.67 and
152.22± 3.93, respectively (p < 0.01 and 0.05); serum triglycerides level by 64.79±3.41 and 67.82±2.26 respectively (p < 0.01
and 0.05); serum low density lipoprotein (LDL) level by 51.58 ± 3.24 and 53.83 ± 1.38, respectively (p < 0.01 and 0.05); blood
glucose level 90.02 ± 0.98 and 92.31±1.63 (p < 0.01 and 0.05) whereas oral administration of 100 mg/kg daily it was showed
no significant (p > 0.05) on serum cholesterol, triglyceride, LDL and blood glucose levels.
© 2011 Universal Research Publications. All rights reserved
Key words: Dalbergia latifolia bark, cholesterol, low density lipoprotein, hypolipidemic, β- sitosterol.
Introduction
Hyperlipidemia and reduced high-density lipoproteins (HDL-
C) ocures a several risk factors that may be life style, geneti,
metabolism or other conditions that enfluence plasma
lipoprotein metabolism [1]. Atherosclerosis is one of the
major risk factors for coronary heart disease and it is widely
recognized that the oxidative modification of human low
density lipoprotein may play an important role [2]. Excess
LDL in the artery wall due to hypercholesterolemia can
undergo oxidative modifications. The oxidative modification
hypothesis of atherosclerosis predicts that low-density lipoprotein oxidation is an early event in atherosclerosis and
that Ox-LDL contributes to atherogenesis. Diabetes mellitus
is chronic metabolic disorder, mainly characterized by
disruptions in carbohydrates, proteins and fat metabolism
caused by the complete or relative insufficiency of insulin
secretion and/or insulin action [3]. Herbal remedies are
apparently efficient, produce least or no side effects in
clinical experience and are comparatively of low costs as
compared to oral synthetic antidiabetic agents [4].
Dalbergia latifolia (Roxb) Family-Fabaceae, a large glabrous
tree a single stem with characteristic smells [5]. The bark is
grey, thin with irregular short cracks, exfoliating in fibrous
longitudinal flakes [6]. It is distributed in Bihar, Bundelkhand
and Central India [7]. It contain dalbinol a new 12a-
hydroxyrotenoid [8], sisafolin coumarin from seeds, β-
sitosterol, also contain dalbergichromene, lupeol, latifolin,
and dalbergin from bark of the tree [9], heartwood contains
latinone, neoflavonoid dalcriodon [10] and Latinone, a substituted phenanthrene-1,4-quinone was isolated from
Dalbergia latifolia [11]. Ethanomedicinally, the stem bark
contain tannin is used for treatment of leprosy and worm [7].
Many species of Dalbergia are important timber trees, valued
for their decorative and often fragrant wood, rich in aromatic
oils [12, 13]. Traditionally various species are reported to be
used as aphrodisiac, abortifacient, expectorant, anthelmentic,
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50 International Journal of Natural Products Research 2011; 1 (4):49-54
antipyretic, appetizer, allays thirst, vomiting, burning
sensation, cures skin diseases, ulcers, diseases of the blood,
reduces obesity, used in leucoderma, dyspepsia, dysentery,
for diseases of the eye and nose, syphilis, stomach troubles,
leprosy, leucoderma, scabies and ringworm [14].
Materials and methods
Chemicals
Diethyl ether, trichloroacetic acid, Disodium ethylene
diamine tetra acetate, Total cholesterol (TC), triglyceride
(TG), high density lipoprotein (HDL), low density lipoprotein
(LDL), glucose, Aspartate aminotransferase (AST), alanine
aminotransferase (ALT), Blood urea nitrogen (BUN) and
creatinine test kits were purchased from clinical chemistry
division of span diagnostic Ltd. India.
Collection and preparation of plant extract
Dalbergia latifolia bark (DLB) was collected in the month of June from area old Delhi market and identified from the
Jamia Hamdard University New Delhi India (reference
voucher No: 001783). Barks were air dried at room
temperature for 3 weeks to get consistent weight. Two
hundred grams of crude powder of barks were shaken with
hydro alcoholic for 24 hrs on an orbital shaker at room
temperature. The extract was resuspended in the respective
solvent [15].
Animals
The female Sprague-Dawley rats (SD rats) were purchase from the Central Drug Research Institute Lucknow (India).
They were housed for 1 week under a 12/12 h light/dark
cycle in a temperature and humidity-controlled room and
freely fed standard laboratory chow with water ad libitum.
The standard laboratory chow contained. All studied were
performed as per CPCSEA guideline, India (Reg.
No.1213/ac/08/CPCSEA/IU).
In vivo hypolipidemic effects of the dried DL-extracts
Thirty-six female Sprague-Dawley rats weighing 125–135 g
were used in this study. The experimental protocol for animal experiments was approved by the Animal Ethic Committee,
Faculty of Pharmacy, Integral University Lucknow. Six rats
per cage were housed in cages in an animal room at the
Faculty of Pharmacy, Integral University. Rats were fed with
commercial pellet diet (National Institute of Nutrition,
Hyderabad, India) and tap water ad libitum. DL-extracts at a
dose of 100, 200, 400 mg/kg and 5 mg/kg standard
(Atorvastatin) drugs daily administered orally using animal
feeding needles at the same time of day for a total 60 days.
Briefly animals were divided into normal control (Group-I)
administered laboratory diet, control obese group induced
with high fat diet [HFD (National Institute of Nutrition, Hyderabad, India)] (group-II), standard drug plus HFD rats
(group-III) and DL-extract (100 and 200 and 400 mg/kg) plus
HFD rats (group-IV, V and VI). At the end of study, rats
were anesthetized with diethyl ether, and blood was collected
by tail vein into glass tubes with Na2EDTA.
Observation and examination method
In-vivo estimation of lipid profile
Rats were anaesthesias with diethyl ether the blood sample
was collected from tail vein at the end of the experiment. The
serum level of TC, HDL, LDL and VLDL were estimating
[16, 17] by methods and TG was estimated by [18] method.
In-vivo effects on serum glucose, blood urea nitrogen and
creatinin of DL-extract At the end of experiments rats were anesthetized with diethyl
ether and blood was collected by tail vein into append drop
tubes and estimation of Blood urea nitrogen and serum
creatine were commercially available span diagnostic kits [19] and glucose was estimated by [20] method.
In-vivo effects on AST and ALT levels of DL-extract
At the end of experiments rats were anesthetized with diethyl
ether and blood was collected by tail vein into append drop
tubes and estimation of AST and ALT were perform
according to the [21] method.
Results and discussion
In-vivo hypolipidemic effects of DL-extract
In the period of induction of hypercholesterolemia through feeding high fat diet in rats, serum cholesterol level increased
over the time period in all groups due to the growth of rats.
The 8 weeks of the induction period, rats in the high fat diet
group exhibited significantly increased serum cholesterol
level, as compared with rats in normal control group
(163.04±2.65 mg/dl p < 0.01) group-I. The administering 5
mg/kg of atorvastatin, used as a positive control, in
hypercholesterolemic rats significantly decreased the levels
of cholesterol (117.22±4.39), HDL (72.87±3.46), LDL
(50.64±3.97), VLDL (9.96±0.79) and TG (64.09±3.29) in
group-III. The administering 200 and 400 mg/kg daily of DL-extract in hypercholesterolemic rats significantly (p<0.01 and
0.05) decreased serum cholesterol level 159.41±1.76 and
152.22±3.93 respectively after 8 weeks of treatment in group
IV-V (Figure 1 & 2). On the other hand, treatment in group
VI with DL-extract 100 mg/kg did not showed significant (p
> 0.05) hypercholesterolemic (159.41±1.76) and
hypotriglyceridemic (71.75±2.16) effect as compared with
hypercholesterolemic rats. Recently, a number of clinical
studies suggest that the increased risk of coronary heart
disease is associated with a high serum concentration of TC,
LDL-C and triglyceride. The abnormally high concentration
of serum lipids is mainly due to the increase in the mobilization of free fatty acids from the peripheral depots
51 International Journal of Natural Products Research 2011; 1 (4):49-54
Figure 1. Effects of DL-extract on serum TC, HDL, LDL
and VLDL levels
Figure 2. Effects of DL-extract on serum triglyceride level
Figure 3. Effects of DL-extract on serum glucose, blood
urea nitrogen and creatinin level.
Figure 4. Effects of DL-extract on serum AST and ALT
levels
52 International Journal of Natural Products Research 2011; 1 (4):49-54
[22]. On the other hand, low serum concentration of HDL-C
is also responsible for coronary heart disease [23]. Recently,
a number of clinical studies suggest dyslipidemia as one of
the major risk factors for coronary disease and preclinical
observations demonstrate that hypercholesterolemia promotes
accumulation of oxidative modified low density lipoprotein (Ox-LDL) in the arterial wall, promoting endothelial cell
(EC) dysfunction and the development of atherosclerosis [24,
25]. In the present study, atorvastatin was used as a positive
control because it is a potent hypolipidemic drug with known
mechanism of action and effects including inhibition of
HMG-CoA reductase, the rate-limiting step in cholesterol
biosynthesis, and resultant increases in LDL-receptors. In
addition, it possesses an inhibitory effect on the inhibition of
LDL oxidation [26, 27]. The treatment of
hypercholesterolemic rats with 200 and 400 mg/kg of DL-
extract along with high fat diet loading for 8 weeks
significantly decreased serum cholesterol, HDL, LDL, VLDL and triglycerides levels but not at 100 mg/kg. Since
phytoconstituent β-sitosterol compound have been reported to
possess hypocholesterolemic effect in-vivo, it may be
speculated that the hypolipidemic effects of DL-extract are
attributed to these compounds. β-sitosterol, a plant sterol
which is structurally similar to cholesterol except for the
substitution of an ethyl group at C24 of its side chain, has
been suggested to reduce cholesterol by lowering the
concentration of LDL cholesterol [28]. The cholesterol level
decreased significantly in liver and plasma without any side
effects in rats fed with the mixture of 0.25% β-sitosterol in diet for days [29]. Thus, a result of the decrease in bile acids
content, cholesterol biosynthesis in rat hepatocytes was
enhanced. Treatment with the DL-extracts (200 and 400
mg/kg) daily result, significantly (p < 0.05) reduced in serum
HDL level but 100 mg/kg did reduced. Hypercholesterolemia
is one of the risk factors of atherosclerosis, in which Ox-LDL
plays a crucial role. In addition, changes of lipid composition
in LDL may also influence the oxidation process. Reduction
of cholesterol and triglycerides contents in LDL may have the
beneficial effect of a decrease in LDL oxidation [30]. The
increased susceptibility of LDL oxidation may be due to changes of triglycerides and cholesterol compositions in rat
LDL particles after induction of hypercholesterolemia.
In-vivo effects on serum glucose, blood urea nitrogen and
creatinin of DL-extract
Glucose, blood urea nitrogen (BUN) and serum creatinin was
found to be significantly increased (101.60±3.16, 48.39±2.17
and 2.06±0.17) when compare with normal control group.
Treatment of 5mg/kg/day atorvastatin showed significantly
decrease (p<0.01) in glucose, BUN and creatinin
(85.93±2.37, 35.28±1.48 and 1.03±0.17 mg/dl) levels in
group III. On treatment of DL-extract 200mg/kg and 400mg/kg daily showed significantly decrease (p<0.01 and
p<0.05) in serum glucose, BUN and creatinin (90.02 ± 0.98,
41.92 ± 1.74 and 1.37 ± 0.04; 92.31±1.63 and 44.63±2.13,
1.46±0.02) in groups V and VI (Figure 3). When
administration of 100mg/kg daily administered, observed non
significant of glucose, BUN and creatinine (98.67 ± 1.43,
49.52±1.74 and 1.76 ± 0.02) in groups IV and compare with obese control group-II. The hypoglycaemia is characterized
by a reduction in insulin-mediated glucose disposal in type-2
diabetes patients [31]. In HFD group more food intake blood
glucose level high as compare to the normal group. The
administration of DL-extract (200 and 400 mg/kg/day) have
been significantly decreased (p<0.01 and p<0.05) the glucose
in circulating blood. The serum creatinine and blood urea
nitrogen levels were impaired due to abnormal regulation,
including elevated glycosylated protein tissue levels and
homodynamic changes within the kidney tissue, and
increased oxidative stress [32]. The HFD induced SD rats
exhibited significantly higher serum creatinine and blood urea nitrogen levels compared to the control normal group.
However, the DL-extract supplement (200 and 400 mg/kg)
daily significantly (p<0.01 and p<0.05) reduced the serum
level of creatinine and blood urea nitrogen in HFD groups V
and VI. Thus, it would appear that the DL-extract supplement
lowered the serum creatinine and blood urea nitrogen levels
by enhancing the renal function that is generally impaired in
diabetes.
In-vivo effects on serum AST and ALT levels of DL-extract
The significantly increase (p<0.01) in the serum AST and ALT levels when compare to normal control group.
Treatment of 5mg/kg/day atorvastatin showed significantly
decrease (p<0.01) in AST and ALT (87.36±3.54 and
32.51±3.98 U/ml) levels in group III. On treatment of DL-
extract (200 and 400 mg/kg) daily showed significantly
decrease (p<0.01 and p<0.05) in serum AST and ALT
(104.79±0.93, 54.17±0.90 and 108.13 ± 1.34, 58.28 ±0.78) in
groups V and VI (Figure 4). When administration of
100mg/kg daily administered, observed non significant of
serum AST and ALT (118.21±0.74 and 65.26±1.34) in
groups IV when compare with obese control group-II. The enzymes AST and ALT are present with higher
concentrations in the liver under normal conditions whereas
during hepatic necrosis or membrane damage, these enzymes
will be released into the systemic circulation, as indicated by
elevated serum enzyme levels [33]. AST is an enzyme that is
present in high quantities in the cytoplasm and mitochondria
of liver, also present in the heart, skeletal muscle, kidney.
ALT is a hepatospecific enzyme that is principally found in
the cytoplasm [34]. These results indicated that DL-extract
have liver protective effect as they significantly reduced
(p<0.01 and p<0.05) the level of both the enzymes on
administration of DL-extract (200 and 400 mg/kg) daily in HFD groups V and VI.
53 International Journal of Natural Products Research 2011; 1 (4):49-54
Conclusion
Thus to conclude, the study showed that administration of
DL-extract at dose level 200mg/kg daily is effective as
hypolipidemic and hypoglycaemic activity. The active
ingredient present in plant may recover the disorders in lipid metabolism noted in hyperlipidemic state and further work
would be necessary to evaluate the active constituents
responsible for the activity and mechanisms of these effects.
Acknowledgement
Authors are very thankful to Honb’le Vice Chancellor,
Integral University, Lucknow for providing research facilities
in university premises for research.
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Source of support: Nil; Conflict of interest: None declared