Acta Scientific MICROBIOLOGY (ISSN: 2581-3226)
Volume 5 Issue 12 December 2022
Biochemical Composition and Effects of Aqueous Extracts of the Leaf, Stem
and Root of Securidaca longipedunculata Fresen (Violet Tree)
on Alloxan Induced Diabetic Rats
Ogbonna AI1*, Madu JM2, Onyimba IA3, Ogbonna USA4, Ejembi EP1,
Nwibari BMW5, Tanko JS6, Peter MK7, Ogbonna CIC1, Azi S8 and Damen
MT1
1
Department of Plant Science and Biotechnology, University of Jos, Nigeria
2
National Biotechnology Development Agency, Lugbe, Airport Road, Abuja, Nigeria
3
Department of Science Laboratory Technology, University of Jos, Nigeria
4
Department of Applied Microbiology and Brewing, Nnamdi Azikiwe University,
Awka, Nigeria
5
Department of Zoology and Environmental Biology, University of Calabar, Nigeria
6
Department of Science Laboratory Technology, Federal College of Forestry, Jos,
Nigeria
7
Department of Forestry Technology, Federal College of Forestry, Jos, Nigeria
8
Department of Pharmacology, University of Jos, Nigeria
*Corresponding Author: Ogbonna AI, Department of Plant Science and
Biotechnology, University of Jos, Nigeria.
Research Article
Received: October 10, 2022
Published: November 07, 2022
© All rights are reserved by Ogbonna AI., et
al.
Abstract
Studies were carried out on the effects of aqueous leaf, stem bark and root extracts of Securidaca longipedunculata. Fresen (violet
tree) on diabetic albino rats. The powdered plant parts were extracted using water as solvent and using maceration method. The
qualitative and quantitative analysis of the biochemical components of the plant parts were carried out using standard methods. The
acute toxicity study was conducted using 24 albino rats for each plant part. The effects of aqueous leaf, stem bark and root extracts of
S. longipedunculata (200 mg/kg) on blood glucose in normoglycemic and alloxan diabetic rats were also investigated using 25 albino
wistar rats. Two mls of each of the extracts were administered orally twice daily for 3 weeks after diabetic induction and the blood
glucose level was measured on daily basis using On-call-plus glucometer. The hypoglycemic activity was evaluated by comparing
the initial blood glucose level with that of the treated and standard. The screening of the aqueous extracts of the plant parts for
biochemical composition revealed the presence of some secondary metabolites of pharmacological significance including alkaloids
(15-18.40 mg/kg), cardiac glycosides (23.30-26.10 mg/kg), flavonoids (23.58-31.05 mg/kg), saponins (215.60-270.59 mg/kg) and
tannins (314.76-339.75 mg/kg) with tannins and saponins in larger quantities. The leaf, stem and root extracts were found to have
LD50 value of 490 mg/kg, 693 mg/kg and 693 mg/kg body weight respectively. The plant part extracts showed significant (p ≤ 0.05)
reduction of Blood Glucose Concentration (BGC) of 3.47, 3.96 and 4.03 mmol/dl for leaf, stem bark and root extracts respectively
after the treatment period indicating that S. longipedunculata has hypoglycemic activity.
Keywords: Securidaca longipedunculata; Biochemical Components; Diabetic; Albino Rats; Extracts
DOI: 10.31080/ASMI.2022.05.1171
Citation: Ogbonna AI., et al. “Biochemical Composition and Effects of Aqueous Extracts of the Leaf, Stem and Root of Securidaca longipedunculata Fresen
(Violet Tree) on Alloxan Induced Diabetic Rats”. Acta Scientific Microbiology 5.12 (2022): 12-22.
Introduction
Diabetes mellitus is a chronic disorder that occurs when the
body is unable to produce or respond to insulin, a hormone that
allows blood glucose to enter the cells of the body and generate
the body’s energy. Diabetes as a metabolic disorder occurs when
the pancreas fails to produce enough insulin or when the body
cells could not respond to the insulin produced by the pancreas
and maintains blood sugar level over a prolonged period [1].
Currently diabetes affects 422 million people worldwide [2] which
shows that approximately 3% of the world population have been
affected by the disease. In 2015, “diabetes was the direct cause
of 1.6 million deaths in the world and about 425 million people
have diabetes in the world” [3]. It was also reported that in 2015,
about 2.0% of the adults in Nigeria have diabetes. This amounts
to about 1,702,900 cases of diabetes. “Symptoms of high blood
sugar include frequent urination, increased thirst and increased
hunger” [4]. Diabetes if not treated well can cause many other
complications. The symptoms of untreated diabetes as reported by
Cooke and Plotnick [5] included “weight loss, polyuria (increased
urination), polydipsia (increased thirst) and polyphagia (increased
hunger) and symptoms may develop rapidly (weeks or months) in
type 1 diabetes mellitus, while they usually develop much more
slowly and may be subtle or absent in type 2 diabetes mellitus”. The
authors also reported that several other signs and symptoms can
mark the onset of diabetes and include blurry vision, excessive loss
of weight, headache, fatigue, itchy skin and slow healing of wounds
or wounds refusing to heal. However, prolonged high blood glucose
can cause the lens of the eye to absorb glucose, resulting in changes
in its shape and changes in vision. Skin rashes that occur in diabetic
conditions are collectively known as diabetic dermadromes [6].
Securidaca longipedunculata (Family: Polygalaceae) is a plant
with several uses in African traditional medicine for treating
diseases such as various sexually transmitted infections, snake bites,
hernias, coughs, fever, anti-helmintic, inflammation, arthritis, skin
infections and others. it is also employed as an aphrodisiac for men
[7]. The Iringa people of Tanzania employ it for the management
of some non-insulin dependent diabetes. S. lonpipedunculata is
known in Nigeria with common names like; violet tree (English),
Uwarmaganigunar (Hausa) which means mother of all drugs,
Ezeogwu (Igbo) and Ipeta (Yoruba). The plant is a medium sized
violet tree, growing up to 6-12 m height, with a characteristic pale
smooth stem bark. Leaves of S. longipedunculata are of varying
sizes and shapes, alternate in arrangement and sometimes spinetipped. The plant presents fine hairs at early stage which they lose
when they are matured. Flowers are in short bunches are always
pink or purple in colour and are sweet scented [8].
Plate 1: Securidaca longipedunculata.
The uses of this plant, Securidaca longipedunculata include a
great variety. There are several uses of the plant both medically
and otherwise around African continent. It was reported that it
can be used to treat ailments as little as headaches or as severe
as inflammatory conditions. It has been known to possess
molluscicidal properties as a result of its saponins component.
The plant has pesticidal activity against beetles in stored grains.
This could be very helpful for small-scale subsistence farmers in
Africa who cannot afford expensive synthetic pesticides. Ojewole
[7] reported that “the roots of the tree can be used for treatments
of human ailments such as coughs, chest pains, toothaches, fevers,
constipation, diabetes and microbial infections. It also possesses
anti-inflammatory properties that help to reduce arthritis pains”.
It has been reported that the methanol extract and the methyl
salicylate component of the roots of S. longipedunculata produce
a potent fish poison and the poison is also used on arrows for
hunting in the wild [9]. Yang., et al. [10] reported that “The
genus Securidaca comprises about 80 species, characterized by
papillionaceous purplish flowers which produce compounds
known as securixanthones with antimicrobial and antioxidant
properties”. Diabetes is eating deep into the society and there is
13
Biochemical Composition and Effects of Aqueous Extracts of the Leaf, Stem and Root of Securidaca longipedunculata Fresen (Violet Tree) on
Alloxan Induced Diabetic Rats
Citation: Ogbonna AI., et al. “Biochemical Composition and Effects of Aqueous Extracts of the Leaf, Stem and Root of Securidaca longipedunculata Fresen
(Violet Tree) on Alloxan Induced Diabetic Rats”. Acta Scientific Microbiology 5.12 (2022): 12-22.
urgent need to search for novel antidiabetic drugs from medicinal
plants or herbal sources. The existing synthetic drugs are now
developing resistance and side effects due to long-term use.
Scientists are in search of bioactive compounds that could aid in
the development and production of novel drugs with hypoglycemic
effects that may control diabetes with little or no side effects. The
paucity of information on the use of S. longipedunculata and the
claims by herbal medical practioners spurred the authors to carry
out this research work.
The main aim of this study was to evaluate various biochemical
constituents and hypoglycemic effects of aqueous leaf, stem bark
and root extracts of Securidaca longipedunculata on animal models.
Materials and Methods
Determination of the biochemical composition of the extracts
Determination of the qualitative biochemical composition of
the extracts
Phytochemical screening of the extracts was done employing
standard methodologies. The tests carried out included those for
alkaloids, cardiac glycosides, saponins, tannins and flavonoids,
anthraquinones, steroids, and carbohydrates [11-14].
Determination of the quantitative biochemical composition of
the extracts
Determination of alkaloids content
The total alkaloids content of the aqueous extract of S.
longipedunculata was determined according to the modified
method of Biradar and Racheti [15]. A weight of 5 g of each sample
was added to 50 ml of a solution containing 10% acetic acid in
ethanol and were mildly stirred for 48 hours. The resultant mixture
was filtered. The extracts were concentrated to one-quarter of the
original volume and 2 ml of 3% sulphuric acid and 8 ml of water
were added to bring the pH to 2.5. This solution was transferred to a
separator funnel and a volume of 10 ml of petroleum ether: diethyl
ether (1:1) solution was added. The bottom phase was collected
and added to ammonium hydroxide solution until precipitate was
completely formed (pH 8.0). The solution was allowed to settle and
the precipitated phase was collected and washed severally with
ammonium hydroxide and chloroform. The precipitated phase was
dried first with sodium sulphate and was then completely dried
by rotavapor and weighed to estimate the percentage of alkaloids
obtained.
Determination of tannins content
About 2 g of each sample was extracted for 20 hours with
anhydrous ether. The resultant residue was boiled in 300 ml water
for 2 hours then cooled. It was diluted to 500 ml and filtered. About
25 ml volume of this infusion was measured into 2 L porcelain
dish; a volume of 20 ml indigo solution was added and also 750 ml
water. One ml of standardized potassium permanganate solution
was added gradually at a time until the blue solution changes to
green. Then few drops of standardized potassium permanganate
solution were added more at a time until solution becomes golden
yellow. The mixture of 20 ml Indigo solution was then titrated with
750 ml water; difference in two titrations was multiplied to obtain
Quercitannic acid [16].
Determination of flavonoids content
A weight of 1 g of each of the sample was weighed and repeatedly
extracted with 100 cm3
of 80% methanol at room temperature. The
mixture was then filtered through into a 250 cm3
beaker using filter
paper. The filtrate was evaporated to dryness using water bath at
60o
C. The resultant extracts were weighed and the percentage (%)
flavonoids were calculated [17].
Determination of cyanogenic glycosides (cyanide) content
using alkaline titration method
No. 20 sieve was used to sieve about 10-20 g portion of each
ground sample into 80 ml Kjedahl flask. A volume of 200 ml
distilled water was added and was allowed to stand for 2 to 4 hours
(Autolysis was conducted with apparatus completely connected
for steam distillation). A volume of 150 to 160 ml distillate was
collected in NaOH solution (0.5g in 20 ml water), and then diluted to
definite volume (250 ml). About 100 ml of the distillate was titrated
by adding 8 ml of 6M NH4
OH. Also add 2 ml of 5% KI solution and
titrate with 0.02M AgNO3, using micro burette. Permanent turbidity
was observed especially against black background [18].
Determination of saponins content
The percentage yield (%) of the total saponins content was
determined by gravimetric method as described by Kaur., et al.
[19]. A weight of 1 g in 10 ml of methanolic extract of each plant
part was macerated for 24 hours and then partitioned in a water
and n-butanol (1:1 ratio) solution. The solution was poured into a
separator funnel and was allowed to stand for 2 hours. The upper
n-butanol layer was separated and the solvent was evaporated to
obtain crude saponins extract which was measured and calculated.
14
Biochemical Composition and Effects of Aqueous Extracts of the Leaf, Stem and Root of Securidaca longipedunculata Fresen (Violet Tree) on
Alloxan Induced Diabetic Rats
Citation: Ogbonna AI., et al. “Biochemical Composition and Effects of Aqueous Extracts of the Leaf, Stem and Root of Securidaca longipedunculata Fresen
(Violet Tree) on Alloxan Induced Diabetic Rats”. Acta Scientific Microbiology 5.12 (2022): 12-22.
Collection of drugs and other chemicals
Alloxan monohydrate was purchased from Sigma Co., USA,
Metformin (Glucophage 500 mg) was procured from a local
pharmacy shop in Jos, Plateau State, Nigeria. The other chemicals
such as the solvents and reagents used were of analytical grade
obtained from reputable vendors.
Collection and processing of the plant materials
The preparation of the plant materials was done according to
the methods of Okeke and Elekwa [20]. The fresh leaves, stem and
roots of Securidaca longipedunculata were collected during the
month of August, 2021 between the hours of 7am-10am GMT at
Shere Hills, Jos, Plateau State, Nigeria. The plant was identified
and authenticated by Mr. J. J. Azila in the Herbarium of Federal
College of Forestry, Jos, Plateau State, Nigeria. The plant materials
were properly dried. The dried samples were then pulverized
to powdered forms. Two grammes of each of the samples were
weighed into separate beakers containing 10 mls of distilled water
and were stirred properly. The beakers were left to stand under
room temperature for 24 hours. They were filtered with Whatman
No 1 filter paper to obtain an aqueous solution. The resultant
crude extracts were stored in clean airtight sample bottles in a
refridgerator (4o
C) prior to use. Fresh crude extracts were prepared
each two days.
Determination of acute toxicity (LD50) of the extracts
The acute toxicity (LD50) of the extracts were carried out using
the method of Lorke [21]. For each of the plant part extract, twentyfour albino rats were used for the acute toxicity test in determining
the LD50 in two phases. The initial phase employed twelve rats
which were divided into four groups of 3 rats each. The first group
that was given distilled water only served as control. Groups 2,
3 and 4 were orally treated with 10, 100 and 1000 mg/kg body
weight of the extract respectively. Each of the rats was given a
dose after 5 days of adaptation and acclimatization. The rats were
observed for 24 hours for any mortality. In the second phase, the
rats were grouped into four of one rat each and treated (orally)
with the extract at varying doses (200, 400, 600 and 800 mg/kg)
per body weight. The animals were observed for any death within
24 hours and the final LD50 value was determined and recorded.
Experimental animals and study protocol
Twenty-five albino rats (Wister stock) were obtained from Animal
unit of Department of Pharmacology, Faculty of Pharmaceutical
Sciences, University of Jos, Nigeria. The animals were maintained
under standard laboratory conditions (temperature 25 ± 2o
C; RH 50
± 5%; 12 hrs light/dark cycle) and were fed on normal rodent diet
specially prepared from chick Grower’s mash and were given water
ad libitum throughout the study period. All rats were acclimatized
for 4 days in laboratory conditions before start of experiment. As
at the starting of the experiment, animals’ weights were taken and
they ranged between 100-150 g.
The experiment was designed to access the effects of 200 mg/kg
body weight of aqueous leaf, stem and root extracts of Securidaca
longipedunculata on alloxan diabetic rats. The animals were divided
into five groups (A-E) with each group containing five animals.
Group A (control) were treated with only distilled water. Group
B (control) were induced with alloxan (150 mg/kg body weight)
and treated with metformin (100 mg/kg). Groups C, D and E were
induced with alloxan and treated with leaf, stem and root extract
(200 mg/kg body weight respectively) via gavage technique (oral
route) for 3 weeks [22]. All treatments were carried out up to 3
weeks (21 days). The animals in the first group were administered
water simultaneously for 3 weeks.
Collection and analysis of blood samples for blood glucose
concentration
At the end of the treatment period, the concentration of
blood glucose of the diabetic rats was determined employing the
method of Baker., et al. [23]. The tail of each of the rat was cut at
the tip aseptically with the aid of a new sterile razor blade after
swabbing with methylated spirit. Drop of blood from the tip of
the tail was placed on the sensor of the blood glucose test strip.
The test trip was inserted into the on-call-plus glucometer with an
arrow indicating ready for use. The readings for the Blood Glucose
Concentration were recorded in triplicates. The test was carried
out in the morning by 8 am before the rats were fed after 12 hours
of not eating.
Results
The biochemical composition of plant parts of Securidaca
longipedunculata
The qualitative screening for the biochemical constituents as
shown in Table 1 revealed that the leaf, stem and root of Securidaca
longipedunculata contained saponins, tannins, cardiac glycosides,
alkaloids and flavonoids. Anthraquinones was found only in the leaf
15
Biochemical Composition and Effects of Aqueous Extracts of the Leaf, Stem and Root of Securidaca longipedunculata Fresen (Violet Tree) on
Alloxan Induced Diabetic Rats
Citation: Ogbonna AI., et al. “Biochemical Composition and Effects of Aqueous Extracts of the Leaf, Stem and Root of Securidaca longipedunculata Fresen
(Violet Tree) on Alloxan Induced Diabetic Rats”. Acta Scientific Microbiology 5.12 (2022): 12-22.
and stem while it was absent in the root extract. Carbohydrates and
steroids were completely absent in the three plant parts extracts.
The results of the quantitative screening of the biochemical
constituents of the plant parts extracts as presented in Table
2 showed a significant difference (p ≤ 0.05) amongst the
constituents. The biochemical constituents from aqueous root
extract exhibited highest mean values of S. longipedunculata and
the order of quantitative estimation are total tannins (339.75%)
saponins (270.59%) > flavonoids (31.05 %) > cardiac glycosides
(23.89 %) > alkaloids (15.00%). This was followed by combined
performance of biochemical constituents from aqueous stem bark
extract and the order of increase are total tannins ((315.36 %) >
saponins (238.79 %) > flavonoids (28.78 %) > cardiac glycosides
(26.10 %) > alkaloids (18.40 %). biochemical constituents from
aqueous leaf extract exhibited least trend on increase and the
order of performance are also total tannins (314.76 %) > saponins
(215.60 %) > flavonoids (23.58 %) > cardiac glycosides (23.30 %)
alkaloids (15.03 %).
Acute toxicity (LD50) test
The LD50 value of the plant parts extract was determined at 490
mg/kg, 693 mg/kg and 693 mg/kg for leaf, stem bark and root
extracts respectively as shown in Tables 3a-3c.
Blood glucose concentration
The results of the blood glucose concentration of the
experimental albino rats before and after treatments are presented
in Table 4 showing the mean effect of blood glucose levels of
experimental rats that received different treatments after induction
with alloxan monohydrate including distilled water (Group A),
Metformin (Group B), 200 mg/kg b. w. aqueous leaf, stem and root
extracts (Group C-E) respectively. The results of the mean effects
of blood glucose levels of experimental rats treated with distilled
water thus revealed a significant difference (p ≤ 0.05) amongst the
glucose level when compared with the standard and the highest
value for each level was obtained at 72 hrs after induction with
Alloxan. It was observed that the blood sugar level increased to
6.78 mmol/dL. The treatment with distilled water did not have any
effect on the blood glucose level of the group. The mean effects of
blood glucose levels of experimental rats treated with metformin
(control group) revealed a significant difference (p ≤ 0.05) amongst
the glucose level when compared with the standard and the highest
value for each level was obtained at 72 hrs after induction with
Alloxan (Table 4). After induction, it was observed that the blood
sugar level increased to 7.06 mmol/dL but after treatment the
blood sugar came down to 3.91 mmol/dL.
The mean effects of blood glucose levels of experimental rats
treated with plant leaf extract of S. longipedunculata showed a
significant difference (p ≤ 0.05) amongst the glucose level when
compared with the standard and the highest value for each level
was obtained at 72 hrs after induction with Alloxan the blood sugar
level increased to 7.74 mmol/dL after induction but after treatment
the blood sugar came down to 3.97 mmol/dL. In the same manner,
the mean effects of blood glucose levels of experimental rats
treated with stem bark extract of S. longipedunculata revealed a
significant difference (p ≤ 0.05) amongst the glucose level when
compared with the standard and the highest value for each level
was obtained at 72 hrs after induction with Alloxan. the blood
sugar level increased to 8.22 mmol/dL but after treatment the
blood sugar came down to 3.96 mmol/dL. The mean effects of
blood glucose levels of experimental rats treated with root extract
of S. longipedunculata revealed a significant difference (p ≤ 0.05)
amongst the glucose level when compared with the standard
and the highest value for each level was obtained at 72hrs after
induction with Alloxan. the blood sugar level increased to 7.87
mmol/dL but after treatment the blood sugar came down to 4.03
mmol/dL (Table 4).
Biochemical
Constituent
Leaf Stem Root
Saponins + + +
Tannins + + +
Anthraquinones + + –
Cardiac glycosides + + +
Alkaloids + + +
Flavonoids + + +
Carbohydrates – – –
Steroids – – –
Table 1: Biochemical Constituents of Aqueous Leaf, Stem and
Root Extracts S. longipedunculata.
- = Present – = Absent.
16
Biochemical Composition and Effects of Aqueous Extracts of the Leaf, Stem and Root of Securidaca longipedunculata Fresen (Violet Tree) on
Alloxan Induced Diabetic Rats
Citation: Ogbonna AI., et al. “Biochemical Composition and Effects of Aqueous Extracts of the Leaf, Stem and Root of Securidaca longipedunculata Fresen
(Violet Tree) on Alloxan Induced Diabetic Rats”. Acta Scientific Microbiology 5.12 (2022): 12-22.
Phytochemical
constituents (%) SD ± Mean
Alkaloids 0.19 ± 15.03d
Total Tannins 8.81 ± 314.75a
Saponins 1.09 ± 215.60b
Flavonoids 0.46 ± 23.58c
Cardiac Glycosides 0.49 ± 23.30c
SE 1.99
p-value 0.000
Table 2: Mean Quantitative estimation of some Phytochemical
constituents from the aqueous leaf extract of S. longipedunculata.
Values (in the same column) with the same subscript letters do
not differ significantly from each other according to the Duncan
multiple range test.
(a)
Doses (mg/kg) No of Treated rats No of Dead rats after
24 hrs
Control (Distilled water)
3 0/3
10 3 0/3
100 3 3/3
1000 3 0/3
(b)
Doses (mg/kg) No of Treated rats No of Dead rats after
24 hrs
200 3 0/3
400 3 0/3
600 3 3/3
800 3 3/3
Table 3a: Lethal Dose (LD50) of the Aqueous Leaf Extract of
Securidaca longipedunculata.
n = number of rats per group.
LD50=
LD50 (Oral) = 490 mg/kg
(a)
Doses (mg/kg) No of Treated
rats
No of Dead rats
after 24 hrs
Control (Distilled
water) 3 0/3
10 3 0/3
100 3 0/3
1000 3 3/3
(b)
Doses (mg/kg) No of Treated
rats
No of Dead rats
after 24 hrs
200 3 0/3
400 3 0/3
600 3 0/3
800 3 3/3
Table 3b: Lethal Dose (LD50) of the Aqueous Stem Extract of
Securidaca longipedunculata.
n = number of rats per group.
LD50=
LD50 (Oral) = 693 mg/kg
(a)
Doses (mg/kg) No of Treated
rats
No of Dead rats after
24 hrs
Control (Distilled
water) 3 0/3
10 3 0/3
100 3 0/3
1000 3 3/3
(b)
Doses (mg/kg) No of Treated
rats
No of Dead rats after
24 hrs
200 3 0/3
400 3 0/3
600 3 0/3
800 3 3/3
Table 3c: Lethal Dose (LD50) of the Aqueous Root Extract of
Securidaca longipedunculata.
n = number of rats per group.
LD50=
LD50 (Oral) = 693 mg/kg
17
Biochemical Composition and Effects of Aqueous Extracts of the Leaf, Stem and Root of Securidaca longipedunculata Fresen (Violet Tree) on
Alloxan Induced Diabetic Rats
Citation: Ogbonna AI., et al. “Biochemical Composition and Effects of Aqueous Extracts of the Leaf, Stem and Root of Securidaca longipedunculata Fresen
(Violet Tree) on Alloxan Induced Diabetic Rats”. Acta Scientific Microbiology 5.12 (2022): 12-22.
Blood Glucose Level (mmol/dL)
Treatment
Control Group
(Water)
A
Control Group
(Metformin)
B
Leaf Extract
C
Stem Bark
Extract
D
Root Extract
E
Before induction of
Alloxan
0.07 ± 3.95c 0.06 ± 3.96c 0.25 ± 3.89d 0.24 ± 3.91c 0.08 ± 4.10c
72hrs after induction of
Alloxan
0.06 ± 6.78a 0.11 ± 7.06a 0.32 ± 7.74a 0.67 ± 8.22a 0.16 ± 7.87a
After treatment – 0.22 ± 3.91d 0.36 ± 3.97c 0.11 ± 3.96c 0.10 ± 4.03c
R/standard x conc. of
standard
0.51 ± 5.32b 0.07 ± 5.51b 0.17 ± 5.71b 0.17 ± 5.71b 0.15 ± 5.77b
SE 0.13 0.04 0.13 0.17 0.06
p-value 0.000 0.000 0.000 0.000 0.000
Table 4: Blood Glucose Levels of the Experimental Rats Treated with Plant Part Extract of Securidaca longepedunculata (SD ± n = 5).
Mean values (in the same column) with the same subscript letters do not differ significantly from each other according to the Duncan
multiple range test.
Discussion
The biochemical analyses on the plant, Securidaca
longipedunculata indicated that the plant parts contained various
biochemicals including saponins, tannins, alkaloids, flavonoids,
cardiac glycosides, anthraquinones, though not contained in the
root extract and the absence of carbohydrates and steroids in all the
three extract. The treatment of diabetes with conventional drugs
has been reported to be very expensive and the side effects are
reportedly high. Marella [24] reported that “out of the secondary
metabolites produced by plants, polyphenols, in particular,
flavonoids are suggested as good therapeutic agents in the
management of diabetes mellitus and its chronic complications”.
This report is similar with the findings of this research since
flavonoids (23.30, 26.10 and 23.89 mg/kg) were isolated from the
leaf, stem and root of S. longipedunculata (Table 2). Flavonoids
can lower blood glucose in diabetic rats [25] and it works by the
repairs of damaged pancreatic beta cells and stimulation of insulin
secretion by the pancreatic beta cells. Ajiboye,., et al. [26] reported
that biochemicals obtained from herbal plants could provide an
alternative for the production of new therapeutic agents against the
ailment, diabetes mellitus. These biochemicals include flavonoids,
tannins, terpenoids, cardiac glycosides, alkaloids, and steroids.
Flavonoids have been known to have strong antioxidants potentials.
Ghosh., et al. [27] studied Bacopa monnieri (L.) Wettst. (family:
Scrophulariaceae) a creeping herb found across India. The authors
reported that “Bacosine, a triterpene isolated from ethyl acetate
fraction of B. monnieri showed pronounced reduction in blood
glucose levels in diabetic rats in a dose-dependent manner and that
Bacosine is known to have antihyperglycemic properties rather
than hypoglycemic activity”. They reported further that “Bacosine
works in a way similar to insulin and that its anti-hyperglycemic
activity might be attributed to the increase in the consumption
of peripheral glucose as well as protect against oxidative damage
in alloxan induced diabetes”. Also, “several compounds including
tetracyclic triterpenoid saponins, Bacosides A and B, Hersaponin,
alkaloids viz. Herpestine and Brahmine and flavonoids have been
isolated from the plant, Bacopa monnieri” [27].
Another bioactive phyto-compound is Charantin which a
cucurbitane (triterpenoid) extracted from M. charantia. This
compound has demonstrated antidiabetic activity and is more
effective than tolbutamide which is a standard oral hypoglycemic
drug [28]. Two other Saponin compounds, 3-hydroxycucurbita-5,
24-dien-19-al-7, 23-di-O-β-glucopyranoside and MomordicineII which were extracted from the corolla of M. charantia produce
antidiabetic activity and have also exhibited insulin-releasing
properties in MIN6 β-cells. Al-Amin., et al. [29] in their study
reported that “several active constituents have been isolated from
18
Biochemical Composition and Effects of Aqueous Extracts of the Leaf, Stem and Root of Securidaca longipedunculata Fresen (Violet Tree) on
Alloxan Induced Diabetic Rats
Citation: Ogbonna AI., et al. “Biochemical Composition and Effects of Aqueous Extracts of the Leaf, Stem and Root of Securidaca longipedunculata Fresen
(Violet Tree) on Alloxan Induced Diabetic Rats”. Acta Scientific Microbiology 5.12 (2022): 12-22.
Zingiber officinale (Ginger) including gingerols, the shogaols, as
well as volatile oils of sesquiterpenes, such as β-bisabolene and
monoterpenes, mainly geranial and neral and that 6-shogaol
and 6-gingerol can suppress the development of diabetic
complications”. Other reports indicating anti diabetic effects of
active principles from medicinal plants include those of Shibano
[30] who isolated radicamines A and radicamines B (alkaloids)
Lobelia chinensis which exhibited α-glucosidase activity and also
demonstrated activity for hyperglycemia. From this research work,
the tannins and saponins were reported to be in larger quantities
than other active principles isolated. The results of the research
work were in consonance with the works of Auwal., et al. [31]. The
quantities of tannins and saponins obtained could probably be due
to the solubility of tannins and saponins in aqueous solution as
reported by Tailang and Sharma [32].
The acute toxicity LD50 test showed that Securidaca
longipedunculata was slightly toxic [33] to the laboratory animals
following median lethal dose (LD50) values of 490 mg/kg, 693
mg/kg and 693 mg/kg for the aqueous leaf, stem bark and root
extracts respectively (Table 3a-3c). The results of the acute toxicity
test recorded for this research work was in agreement with the
findings of Auwal., et al. [31]. However, Lork [21] noted that LD50
greater than 5000 mg/kg b. w. is safe. Bulus., et al. [34] in their
work recorded increased symptoms of toxicity on white albino rats
but there was no death at LD50 of 5000 mg/kg b. w. of Terminalia
avicennioides aqueous extract. According to Kagbo and Ejebe [35],
acute toxicity test is a very useful tool in determining the safety or
toxic potential of a natural product or substance that its toxicity
profile has not been elucidated. Yehya., et al. [36] indicated that
“Preliminary toxicological tests are always necessarily conducted
in order to select the safe doses for antidiabetic studies”. They
went further to report that researches on herbal plants provide
window for the evaluation of safety profile of herbal products and
for setting up an avenue for mapping a safe dose for use in humans
and animals.
There is a growing interest on the use of herbal plants as
alternative medical therapies for lowering blood sugar levels
in patients diagnosed with diabetes mellitus (DM). The present
study assessed the effects of aqueous leaf, stem bark and root of S.
longipedunculata on diabetic albino rats. The normoglycemic rats
were induced for diabetes using alloxan monohydrate within 72 hrs.
Alloxan is a known inducer of hyperglycemia solely by destroying
pancreatic β-cells through redox-mediated mechanisms. ‘This
auto-oxidation of glucose in hyperglycemic condition can trigger
lipid peroxidation and changes in antioxidant defense mechanisms
which could lead to dysfunctions metabolism of glucose” [37].
Metformin which was employed as a standard drug is a biguanide
its duty is to reduce glucose synthesis in the hepatic cells and
also boosting of peripheral insulin sensitivity [38]. Metformin
(Glucophage) is an anti-hyperglycemic agent that improves glucose
tolerance by lowering basal and postprandial plasma glucose
levels in type 2 diabetes [39]. The potentials of herbal medicines
in controlling hyperglycemia cannot be over emphasized. Alam., et
al. [40] reported in their work that plants bioactive compounds can
be rely upon as a steppingstone to manufacture new antidiabetic
therapeutics to help in treating diabetes and their associated
complications. “In Vitro preliminary screening of aqueous extract
of the leaves of Securidaca longipedunculata for anti-hyperglycemic
property” was carried out by Onyeche and Kolawale [41]. Our
research findings have demonstrated that the aqueous extract
of the test plant parts brought about significant lowering of the
blood glucose level in the diabetic rats when compared with the
standard. Medicinal effects of these plant parts are as a result of
the presence of biologically active compounds [42,43]. Flavonoids
have been reported to have antidiabetic effects by the enhancement
of insulin secretion and insulin mediated glucose uptake by cells
after regeneration of pancreatic β-cells [24]. Panda [44] reported
that Moringa oleifera leaf alcoholic extract contain bioactive
compounds like flavonoids, alkaloids, tannins, steroids and
glycosides which are effective in treating diabetic complications.
Quercetin and kaempferol are two major bioactive constituents
which were isolated from M. oleifera had found to reduce blood
glucose (33.34%) in diabetic rat models in few weeks [45].
Also there have been reports to support the antidiabetic activity
of Vernonia amygdalina plant parts. Gyang., et al. [46] observed
that chloroform extract of the plant, V. amygdalina has anti-diabetic
effect in both normoglycemic and alloxan-induced hyperglycemic
rats. Other researchers have reported on the use of herbal plants
for hypoglycemic activity [40,42,47-51,53,54]. Diabetes mellitus
especially that of Type 2 is characterized by decreased physical
activity as well as increased sedentary habits, resulting to elevated
systemic inflammation [55].
19
Biochemical Composition and Effects of Aqueous Extracts of the Leaf, Stem and Root of Securidaca longipedunculata Fresen (Violet Tree) on
Alloxan Induced Diabetic Rats
Citation: Ogbonna AI., et al. “Biochemical Composition and Effects of Aqueous Extracts of the Leaf, Stem and Root of Securidaca longipedunculata Fresen
(Violet Tree) on Alloxan Induced Diabetic Rats”. Acta Scientific Microbiology 5.12 (2022): 12-22.
Bibliography
Conclusion
Aqueous leaf, stem and root extracts of Securidaca
longipedunculata effectively lowered blood glucose levels in wister
albino rats induced with diabetes mellitus. However, further
studies are required to elucidate the particular active compound(s)
of the biochemicals that are responsible for the anti-hyperglycemic
effects of the plant and suggests that the active compounds isolated
from the plant used as lead compound for the production of antidiabetic drugs.
Acknowledgements
Authors are grateful to the staff of Animal Unit, Department of
Pharmacology, Faculty of Pharmaceutical Sciences, University of
Jos especially Mr. Lungfa Ishaku Lar for the technical assistance
they provided to us. We are thankful to Mr. Thomas Yakubu for
preparing the aqueous plant part extracts used in the work. We are
especially grateful to Mr. Sunday Azi for preparing all the extract
and drug dilutions used for the work. We are equally thankful
to Dr. BMW Nwibari who ordered for and purchased the alloxan
monohydrate from Sigma Co. USA.
Conflict of Interest
Authors have declared that no competing interests exist.
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22
Biochemical Composition and Effects of Aqueous Extracts of the Leaf, Stem and Root of Securidaca longipedunculata Fresen (Violet Tree) on
Alloxan Induced Diabetic Rats
Citation: Ogbonna AI., et al. “Biochemical Composition and Effects of Aqueous Extracts of the Leaf, Stem and Root of Securidaca longipedunculata Fresen
(Violet Tree) on Alloxan Induced Diabetic Rats”. Acta Scientific Microbiology 5.12 (2022): 12-22.
