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Safety, Efficacy and Toxicological Evaluation of a Novel, Patented Anti-Diabetic Extract of Trigonella Foenum-Graecum Seed Extract (Fenfuro(TM)).

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Abstract Safety and anti-diabetic efficacy of a novel, proprietary Trigonella foenum-graecum seed extract [novel fenugreek extract (FE), Fenfuro(TM), CR0010810) enriched in furostanolic saponins (>60% w/w, HPLC) were assessed. Concerning safety, we undertook studies dealing with acute oral toxicity, 28-day sub-chronic toxicity and Ames' bacterial reverse mutation assay that revealed no toxicity. Concerning efficacy, we examined beneficial effects of the extract on rats with type 2 diabetes (T2D). Male Sprague-Dawley rats received a high-fat diet for two weeks followed by streptozotocin (STZ, 35 mg/kg i.p.) to produce T2D. Seven days post STZ, rats showing > 300 mg/dl fasting plasma glucose level (PGL) were included in the study. FE (150- or 450 mg/kg po) and glipizide (5 mg/kg po) were administered once daily for 20 days and then twice daily for another 10 days (total 30 days). Blood samples were collected at 0, 10, 20 and 30 days of treatment and estimated for fasting plasma triglyceride (PTG), total cholesterol and insulin levels. After 30 days, FE and glipizide-treated diabetic animals were treated in combination with or without metformin (100 mg/kg) twice daily for another 10 days. FE didn't influence body weight, feed and water intake. FE (150 mg/kg po) reduced PTG levels in T2D rats by 22%, 24.6% and 29% at 10-, 20- and 30-days of treatment, respectively, while glipizide (5 mg/kg po) reduced the PTG levels by 57.4%, 46.2% and 39.4% at these time points. FE (450 mg/kg) treatment in STZ-induced diabetic rats produced significant hypoglycemic activity (approximately 31.5%) as compared to insulin (48.2% with 1 U/kg i.p.). FE (150 mg/kg po) and Metformin (100 mg/kg po) combined produced significant reduction (20.7%) of plasma glucose level in T2D rats. No adverse effects were observed. We conclude after extensive in vitro and in vivo safety and efficacy studies that FE is safe and effective in treating T2D.
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ISSN: 1537-6516 (print), 1537-6524 (electronic)
Toxicol Mech Methods, 2014; 24(7): 495–503
!2014 Informa Healthcare USA, Inc. DOI: 10.3109/15376516.2014.943443
RESEARCH ARTICLE
Safety, efficacy and toxicological evaluation of a novel, patented
anti-diabetic extract of Trigonella Foenum-Graecum seed extract
(Fenfuro)
Anand Swaroop
1
, Manashi Bagchi
1
, Pawan Kumar
2
, Harry G. Preuss
3,4
, Kiran Tiwari
2
, Palma Ann Marone
5
, and
Debasis Bagchi
1,6
1
Research and Development, Cepham Inc., Piscataway, NJ, USA,
2
Chemical Resources, Panchkula, Haryana, India,
3
Department of Biochemistry,
Georgetown University Medical Center, Washington, DC, USA,
4
Department of Medicine and Pathology, Georgetown University Medical Center,
Washington, DC, USA,
5
Eurofins/Product Safety Laboratories, Dayton, NJ, USA, and
6
Department of Pharmacological and Pharmaceutical Sciences,
University of Houston College of Pharmacy, Houston, TX, USA
Abstract
Safety and anti-diabetic efficacy of a novel, proprietary Trigonella foenum-graecum seed extract
[novel fenugreek extract (FE), Fenfuroä, CR0010810) enriched in furostanolic saponins (460%
w/w, HPLC) were assessed. Concerning safety, we undertook studies dealing with acute oral
toxicity, 28-d sub-chronic toxicity and Ames’ bacterial reverse mutation assay that revealed no
toxicity. Concerning efficacy, we examined beneficial effects of the extract on rats with type 2
diabetes (T2D). Male Sprague–Dawley rats received a high-fat diet for 2 weeks followed by
streptozotocin (STZ, 35 mg/kg i.p.) to produce T2D. Seven days post-STZ, rats showing
300 mg/dl fasting plasma glucose level (PGL) were included in the study. FE (150- or 450- mg/
kg p.o.) and glipizide (5 mg/kg p.o.) were administered once daily for 20 d and then twice daily
for another 10 d (total 30 d). Blood samples were collected at 0, 10, 20 and 30 d of treatment
and estimated for fasting plasma triglyceride (PTG), total cholesterol and insulin levels. After
30 d, FE and glipizide-treated diabetic animals were treated in combination with or without
metformin (100 mg/kg) twice daily for another 10 d. FE did not influence body weight, feed and
water intake. FE (150 mg/kg p.o.) reduced PTG levels in T2D rats by 22%, 24.6% and 29% at 10,
20 and 30 d of treatment, respectively, while glipizide (5 mg/kg p.o.) reduced the PTG levels by
57.4%, 46.2% and 39.4% at these time points. FE (450 mg/kg) treatment in STZ-induced diabetic
rats produced significant hypoglycemic activity (approximately 31.5%) as compared to insulin
(48.2% with 1 U/kg i.p.). FE (150 mg/kg p.o.) and metformin (100 mg/kg p.o.) combined
produced significant reduction (20.7%) of PGL in T2D rats. No adverse effects were observed.
We conclude after extensive in vitro and in vivo safety and efficacy studies that FE is safe and
effective in treating T2D.
Keywords
28-d sub-chronic toxicity, acute oral toxicity,
Ames’ bacterial reverse mutation assay,
animals, Fenfuro (FE), glipizide, metformin,
type 2 diabetes
History
Received 29 May 2014
Revised 28 June 2014
Accepted 28 June 2014
Published online 11 September 2014
Introduction
Diabetes mellitus or simply diabetes is a metabolic disorder
wherein the body does not produce or adequately use the
hormone insulin leading to a dysregulation of glucose (Bagchi
& Sreejayan, 2012; Brunetti et al., 2014). The disease is
growing at epidemic rate, affecting over 220 million people
worldwide, and is expected to double by 2030. In 2010, 10.9
million people over 65 years of age had diabetes and 215 000
people under the age of 20 years had either type 1 or type 2
diabetes. A review of the data from the 2011 National
Diabetes Fact Sheet, which was released on 26 January 2011,
demonstrated that a total of 25.8 million children and adults
in the United States, which is approximately 8.3% of the
population, have diabetes, while a total of 79 million people
are pre-diabetic (http://www.diabetes.org/diabetes-basics/
statistics/#sthash.KvZJ61P1.dpuf; http://www.diabetes.org/
diabetes-basics/statistics/).
Diabetes has a very large economic impact worldwide
(Bagchi & Sreejayan, 2012; Paneni et al. 2014).
Approximately, 25.8 million people suffer from Diabetes in
USA, 76 million people are diagnosed to be pre-diabetic, and
366 million people around the World suffer from diabetes.
In USA alone, the cost of diagnosed diabetes in 2012 is
estimated at $ 245 billion, including $ 176 billion in direct
medical costs. In 2013, the average diabetes-related expend-
itures per person in the US were $ 9800 (http://www.dia-
betes.org/diabetes-basics/statistics/). The more alarming news
is that about 7 million of those 25.8 million diabetics remain
undiagnosed until they develop a major complication such as
Address for correspondence: Debasis Bagchi, PhD, MACN, CNS,
MAIChE, University of Houston College of Pharmacy, Houston, TX
77204, USA. Tel: 925 948 6951. E-mail: debasisbagchi@gmail.com
Toxicology Mechanisms and Methods Downloaded from informahealthcare.com by University of Georgia on 09/19/14
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stroke, peripheral circulatory disorders, atherosclerosis, neur-
opathy, nephropathy, retinopathy, amputation or blindness,
which further adds to the economic burden of diabetes on
society (http://www.diabetes.org/diabetes-basics/statistics/).
In addition, diabetes is a polygenic disorder, and the
pathogenesis of diabetes involves magnitudes of both genetic
and environmental factors that adversely affect insulin
secretion and tissue response to insulin. Genomic explorations
have attempted to identify genetic variants that may contrib-
ute to the development of diabetes (Brunetti et al., 2014).
A thorough understanding of the intricate mechanistic aspects
of diabetes and its complications will help the scientists/
clinicians to design effective therapeutic strategies to curb the
epidemic (Bagchi & Sreejayan, 2012).
Diabetes is characterized by metabolic dysfunction pri-
marily mediated through carbohydrate metabolism, mani-
fested by hyperglycemia resulting from compromised insulin
secretion or impaired insulin action (Bagchi & Sreejayan,
2012). If untreated, diabetes may lead to a series of
complication affecting vascular systems, eyes, nerves and
kidneys leading to cardiomyopathy, neuropathy, nephropathy,
retinopathy, limb amputation or even mortality (Bagchi &
Sreejayan, 2012).
Type 1 diabetes is a condition wherein the pancreas
produces little or no insulin, whereas type 2 diabetes (T2D),
the more prevalent form, results when the body becomes
resistant to the effects of insulin or does not produce sufficient
insulin (Bagchi & Sreejayan, 2012). A critical appraisal of the
various pharmacological modalities of the management of
diabetes, which includes structurally diverse drugs and
pharmacological agents that targets diverse pharmacological
agents, that targets different mechanisms in the underlying
pathogenesis of diabetes. However, most of these drugs have
serious adverse effects (Brunetti et al., 2014). On the other
hand, scientists have suggested alternative therapies including
balanced diets enriched in fresh fruits, vegetables, dietary
fiber, anti-oxidants and structurally diverse phytopharmaceu-
ticals, in conjunction with regular physical exercise (Bagchi &
Sreejayan, 2012).
Fenugreek (Trigonella foenum-graecum) seeds have
demonstrated novel anti-diabetic efficacy in various experi-
mental model. Studies in animal models have demonstrated
that fenugreek has strong anti-diabetic potential. Furthermore,
human studies also confirmed blood glucose and cholesterol
lowering property of fenugreek seeds and leaves (Elujoba &
Hardman, 1987; Nour & Magboul, 1986; Sauvaire et al.,
1991; Yadav & Baquer, 2014). Fenugreek (Trigonella foenum-
graecum) seeds are high in soluble fiber which helps to lower
blood sugar by slowing digestion and absorption of carbohy-
drate and reduce gastrointestinal absorption of glucose, which
suggests that it might be effective in the treatment of diabetes
(Madar et al., 1988; Sauvaire & Baccou, 1978; Sharma et al.,
1990, 1991, 1996; Yadav & Baquer, 2014). Fenugreek seeds
are also rich sources of vitamins, minerals and anti-oxidants,
which help to protect the cells from free radical-induced
oxidative injury (Bordia et al., 1997; Yadav & Baquer, 2014).
We have developed a novel, water-soluble Trigonella
Foenum-Graecum seed extract, FE enriched in 460%
furostanolic saponins, in our laboratories. We conducted
acute oral toxicity, 28-d sub-chronic toxicity and Ames’
bacterial reverse mutation assay to demonstrate the broad
spectrum safety. The anti-diabetic efficacy of FE was also
investigated in streptozotocin-induced T2D rat model (STZ,
35 mg/kg i.p.) in two different doses (150- and 450- mg/kg
body weight p.o.) over a period of 30 consecutive days.
Efficacy of FE was investigated, individually, and in
combination with glipizide (5 mg/kg p.o.) and metformin
(100 mg/kg p.o.), respectively.
Materials and methods
Preparation of a novel Fenugreek Extract
M
FenfuroÔ(FE) is a unique, water-soluble, patented, novel
supplement derived from Fenugreek (Trigonella foenum-
graecum) seeds (Cepham Inc., Piscataway, NJ). FE is
manufactured using a novel, patented process by solvent
extraction followed by column purification (US Patent
8,217,165).
Animals and treatment
Safety studies including acute oral toxicity and repeated dose
28 d sub-chronic toxicity studies were conducted at Eurofins/
Product Safety Laboratories (Dayton, NJ) and National
Toxicology Center (National Institute of Pharmaceutical
Education and Research NIPER, S.A.S. Nagar, Punjab,
India) in compliance with the Good Laboratory Practices
(GLP) as defined in 21CFR58 by the US Food and Drug
Administration (FDA, 1987) and in accordance with the
Organization for Economic Cooperation and Development
(OECD) guidelines for testing of chemicals (OECD, 1998) and
Testing of Chemicals (No. 407 ‘‘Repeated dose 28-d oral
toxicity study in rodents’’ adopted on 27 July 1995). Ames
bacterial reverse mutation assay was conducted in INTOX Pvt.
Ltd. (Pune, India) in compliance with the OECD Guidelines for
Testing of Chemicals (No. 471, Section 4: Health Effects).
Animals were cared in accordance with the most recent Guide
for the Care and Use of Laboratory Animals DHEW (NIH). All
necessary animal protocol approvals were obtained for these
studies. Detailed animal protocols are provided in individual
toxicological assessments.
For anti-diabetic efficacy, male Sprague–Dawley rats
(140–160 g) were obtained from the central facility of
National Institute of Pharmaceutical Education and
Research (NIPER, S.A.S. Nagar, Punjab, India), maintained
under standard environmental conditions (temperature
20–24 C, relative humidity 50–60% and 12-h light and
12-h dark cycle) and acclimatized to laboratory environment
1 week prior to the study. All the rats were housed in
polypropylene rat cages, fed with rodent chow and drinking
water was purified using aqua guard ad libitum. All the
experimental protocols of the study was approved by the
institutional animal ethics committee (IAEC) and performed
in accordance with the rules and guidelines on animal
experimentation.
Acute oral toxicity
The acute oral toxicity evaluation (Up and Down Procedure)
was conducted in rats and mice to determine the potential of
FE to produce acute oral toxicity from a single dose through
496 A. Swaroop et al. Toxicol Mech Methods, 2014; 24(7): 495–503
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the oral route. The rats and mice were singly housed in
suspended stainless steel cages with mesh floors conforming
to the size recommendations in the most recent Guide for the
Care and Use of Laboratory Animals (National Research
Council, 2011). Litter paper was placed beneath the cage and
was changed at least three times per week. The animal had
free access to standard rodent chow (Purina Rodent Chow#
5012) and filtered tap water ad libitum, and were maintained
at controlled temperature (19–25 C), relative humidity
(30–66%) and light cycle (12 h light/12 h dark). The animals
were acclimated to laboratory conditions prior to initiation
of dosing.
A total of 48 Sprague–Dawley rats (24 male and 24 female;
initial body wright 158–172 g) were obtained from Central
Animal Facility (S.A.S. Nagar, Punjab, India). Male and
female rats were administered a single oral dose of either 0,
500 mg/kg, 1000 mg/kg or 2000 mg/kg by gastric intubation.
The treated animals were observed at regular time intervals
on the day of dosing after the administration of the test item
at about 30 min, 1 h, 2 h, 4 h and 6 h for signs of gross toxicity.
Thereafter, all animals were observed once a day over a
period of 14 d. Cage side observations include changes in
the skin, fur and mucous membranes, and also respiratory
changes. The body weight of each rat was recorded prior
to the dosing on day 1, day 8 and on the terminal sacrifice
day 15. Group mean body weights were calculated. Food and
water intakes were recorded twice a week. For necropsy
analysis, all animals were kept on fasting for approximately
4 h before sacrifice. At the end of the observation period, all
animals were euthanized by CO
2
asphyxiation. All animals
were examined for gross pathological changes, if any.
In another separate and parallel experiment, a total of 48
Swiss albino mice (24 male and 24 female; initial body wright
18–20 g) were obtained from Central Animal Facility (S.A.S.
Nagar, Punjab, India). Male and female mice were adminis-
tered FE in a single oral dose of either 0, 500 mg/kg,
1000 mg/kg or 2000mg/kg by gastric intubation. The treated
animals were observed at regular time intervals on the day of
dosing after the administration of the test item at about 30 min,
1 h, 2 h, 4 h and 6 h for signs of gross toxicity. Thereafter, all
animals were observed once a day over a period of 14 d. Cage
side observations include changes in the skin, fur and mucous
membranes, and also respiratory changes. The body weight of
each mouse was recorded prior to the dosing on day 1, day 8
and on the terminal sacrifice day 15. Group mean body weights
were calculated. Food and water intakes were recorded twice a
week. For necropsy analysis, all animals were kept on fasting
for approximately 4h before sacrifice. At the end of the
observation period, all animals were euthanized by CO
2
asphyxiation. All animals were examined for gross patho-
logical changes.
In another set of experiment, three healthy young adult
female, nulliparous and non-pregnant albino Sprague–Dawley
rats (initial body weight 159–168 g) were obtained from Ace
Animals, Inc. (Boyertown, PA). Female rats were selected for
the test because they are frequently more sensitive to the
toxicity of test compounds than males. FE was orally
administered a single oral dose of 5000 mg/kg body weight
by gastric intubation. Before each dosing, rats were fasted
overnight, examined through the fasting period for health, and
weighed (initial). Individual doses were calculated based on
the specific gravity and concentration of the test mixture.
An initial limit dose of 5000 mg/kg body weight was
administered to one healthy female rat by oral gavage.
At the end of the observation period, all animals were
euthanized by CO
2
asphyxiation. All animals were examined
for gross pathological changes.
All the animals were observed for mortality, signs of gross
toxicity and behavioral changes during the first several hours
post-dosing and at least once daily for 14 d after dosing.
Individual body weights were recorded prior to administration
and again on days 7 and 14 (termination) following dosing.
Necropsies were performed on all animals at terminal
sacrifice. Observations included gross evaluation of skin
and fur, eyes and mucous membranes, respiratory, circulatory,
autonomic and central nervous systems, somatomotor activity
and behavioral pattern. Particular attention was directed to
observations of tremors, convulsions, salivation, diarrhea and
coma. All animals were euthanized by CO
2
inhalation at the
end of the 14-d observation period and gross necropsies were
performed on all animals. Tissues and organs of the thoracic
and abdominal cavities were examined.
Repeated dose 28-d sub-chronic toxicity study
The repeated dose 28-d study was conducted in 20 male and
20 female Sprague–Dawley rats, 5- to 6-weeks-old male
(initial body weight range 176–206 g) and female (initial body
weight range 163–184 g). The animals were acclimatized for
5 d and housed 2–3 rats per sex per cage in sterilized solid
bottom polypropylene cages with stainless steel grill tops,
facilities for food and water bottle and bedding of clean paddy
husks (temperature ranges between 22 ± 3 C; relative humid-
ity 30–70%; illumination cycle 12 h light/12h dark). The
cages were suspended on stainless steel racks. Standard rodent
chow and potable tap water filtered through Aquaguard’s
Reviva Reverse Osmosis System (water filter cum purifier)
was provided ad libitum. After acclimatization and veterinary
examination, the rats were randomly assigned to control and
treatment groups with individual weight of animals not
exceeding ±20% of the average weight in each sex and group
at the beginning of the study.
FE was orally administered at 0, 250, 500 and 1000 mg/kg
body weight/day in a dose volume of 5ml/kg body weight for
28 consecutive days by gastric intubation each day around the
same time. FE is free water soluble and distilled water was
used as a vehicle in all groups. The dosage administered to
individual rat as per the weekly body weight. The subsequent
change in the dose administration was again adjusted after
1 week depending on the change in the body weight.
The control animals were administered with distilled water
as vehicle.
The rats were daily subjected to general cage side clinical
examinations to characterize onset and duration of clinical
signs. All signs of behavioral changes or reaction to treatment
were recorded for individual animals. Clinical signs were
recorded on weekly clinical observation and detailed abnor-
mality observation sheets for individual animals. All cages
were checked twice daily (morning and evening) for dead
animals. Opthalmological examinations were performed for
DOI: 10.3109/15376516.2014.943443 Safety and efficacy of FenfuroÔ497
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all rats prior to the treatment and last week of treatment
period (24th and 25th day of the treatment period for male and
female, respectively). Body weight of each animal was
recorded weekly once during the course of the treatment
and before sacrifice. Feed consumption was recorded weekly
once during the course of the treatment. The measured
quantity of 150 ± 5 g food was offered in each cage and
leftover quantity was measured after 24 h and food intake
per rat was calculated. Water intake was also recorded. Each
animal from control and treatment groups was placed in
digital photoactometer for 5 min and scores were recorded.
The motor activity scores were recorded before treatment and
the last week of post-treatment period.
On completion of 28 consecutive days of treatment and
prior to necropsy, blood samples were withdrawn after
anesthesia from the orbital sinus of male and female rats of
each group. For hematology study, blood was collected with
potassium EDTA and for clinical chemistry blood was
collected with heparin. All animals were fasted overnight
before necropsy.
Hematology parameters including hemoglobin (Hb), total
count, differential count, RBC count, hematocrit (PCV), platelet
count, mean corpuscular volume (MCV), mean corpuscular
hemoglobin (MCH) and mean corpuscular hemoglobin con-
centration (MCHC) were performed using a Hematoanalyzer
MS-9-5 (Melet Schloesing Laboratories, Osny, France).
Analysis was done using in-built veterinary software for various
hematology parameters. The clinical chemistry evaluation was
performed on plasma isolated from blood samples. Plasma was
separated and analyzed for sodium and potassium, glucose,
cholesterol, blood urea nitrogen, creatinine, total protein,
albumin, alanine aminotransferase and aspartate aminotransfer-
ase using the biochemistry semi-autoanalyzer ‘‘Microlab – 300
(MERCK)’’ with kits manufactured by Merck Specialities Pvt.
Ltd, Ambernath, Mumbai, India. Urine samples were collected
by using metabolic cages and appearance/color, glucose,
bilirubin, ketone, specific gravity, blood and pH. Protein and
urobilinogen was assessed using Uriscan strip (YD Diagnostics,
Kyunggi-Do, South Korea).
On completion of 28 d of treatment, rats were sacrificed by
CO
2
asphyxiation. Completed necropsy was carried out on all
animals to score the gross lesions. Tissues including brain,
stomach, large intestine, small intestine, liver, kidneys,
adrenal glands, spleen, heart, thymus, lungs, testis, ovary
and uterus were collected from all animals by necropsy and
preserved in 10% formalin. Individual weights of the liver,
adrenals, spleen, heart, kidney, brain, testes, ovaries and
thymus were recorded after necropsy. Values of these organs
as percent of terminal body weights were estimated (relative
organ weights). The required tissues for histology slide
preparations were embedded in paraffin wax, 5-mm tissue
sections were cut and stained with haematoxylin and eosin.
The histological examination was conducted on the specified
lists of tissues from the control and treated animals.
Mutagenicity test Ames’ bacterial reverse mutation
assay
The Salmonella typhimurium reverse mutation test (Maron &
Ames, 1983, Ames Test) was carried out in compliance with
OECD guidelines for testing of chemicals (No 471, Section 4:
Health effects) on conduct of ‘‘Bacterial Reverse Mutation
Assay’’, adopted on 21 July 1997, and ICH draft consensus
guideline S2 (R1) on Genotoxicity Testing.
FE was evaluated in the Ames Test carried out as a pre-
incubation assay to determine its ability to induce reverse
mutation at selected histidine loci in five tester strains of
S. typhimurium, namely TA 1535, TA 97a, TA 98, TA 100 and
TA 102 in the presence and absence of a metabolic activation
system (S9) (Clive et al., 1983). Analytical grade water
was used as a vehicle. Following the preliminary solubility/
precipitation test and a cytotoxicity test, five tester strains
were exposed to the test article in triplicate cultures at the
doses of 5000, 1500, 500, 150 and 50 -mg/plate in the presence
and absence of an exogenous metabolic activation system
(S9 mix).
Anti-diabetic efficacy of FE in type 2 diabetic
rat model
Male Sprague–Dawley rats were fed high-fat diet (HFD; 58%
fat on calorie basis) for the duration of 2 weeks followed by
low dose of streptozotocin (STZ, 35 mg/kg, i.p.) dissolved in
citrate buffer (pH 4.4) to induce diabetes, while normal
control group rats were fed on normal pellet diet. Control rats
(n¼6) received only citrate buffer (1 ml/kg, i.p.). All the
animals were checked for plasma glucose level (PGL) 2 d
post-STZ injection to confirm diabetes. The rats having
300 mg/dl of PGL were only considered diabetic and
included in the study. The body weight was regularly
monitored.
The diabetic animals were randomly divided into four
groups consisting each of 12 rats post 1 week STZ/vehicle
injection based on their mean 6 h fasting plasma triglyceride
(PTG) and plasma total cholesterol (PTC) before FE and
glipizide treatment, while the normal control rats were
injected with citrate buffer only. FE at two different doses
(150- and 450 mg/kg body weight, p.o.) and glipizide (5mg/
kg body weight, p.o.) were administered in diabetic rats once
daily for 20 consecutive days and then continued twice daily
for 10 d, totally over a period of 30 d, while the respective
control rats were given vehicle distilled water 2 ml/kg body
weight. Body weights, feed (g/d) and water intake (ml/d) were
monitored at regular intervals. Blood samples were collected
at 0-, 10-, 20- and 30- d of treatment for fasting PTG and PTC
using commercially available colorimetric diagnostic kits
from Accurex Biomedical Pvt. Ltd, Thane, India. The
remaining plasma samples were stored at 20 C until the
plasma insulin (PI) was determined using ELISA rat insulin
kit (Linco Research, St Charles, MO).
Effect of chronic treatment of combination therapy
(FE ± metformin) on plasma glucose level in
diabetic rats
In order to evaluate the effect of combination therapy
(FE ± Metformin), all the diabetic animals at the end of 30 d
of study were randomly re-grouped (within each respective
vehicle-treated group) in such a way that each group
consisting of six animals (total nine groups as mentioned in
the Table 3). FE at two different doses (150- and 450- mg/kg
498 A. Swaroop et al. Toxicol Mech Methods, 2014; 24(7): 495–503
Toxicology Mechanisms and Methods Downloaded from informahealthcare.com by University of Georgia on 09/19/14
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body weight, p.o.) and glipizide (5 mg/kg body weight, p.o.)
were continued with or without metformin (100 mg/kg p.o.)
twice daily for the next 10 d. Blood was collected for
estimating 6 h fasted PGL to evaluate the effect of different
combination treatment strategies among various treatment
groups and vehicle-treated groups.
Statistical analysis
The results are expressed as mean ± S.E.M. All the data of
different treated diabetic groups as compared to the control
groups were analyzed using one-way analysis of variance
followed by Tukey’s post-hoc test. A value of p50.05 was
considered statistically significant.
Results
Acute oral toxicity
Acute oral toxicity of FE was extensively investigated
in Sprague–Dawley rats and Swiss Albino mice following
Up and Down procedure. FE did not induce any mortality
or gross toxicity at 500, 1000 and 2000 mg/kg body weight
dose in male and female Sprague–Dawley rats or Swiss
Albino mice. No adverse pharmacological effects or abnormal
behavior was observed. At a higher dose level of 5000 mg/kg
body weight, FE did not cause any mortality and did not
demonstrate any signs of gross toxicity, adverse pharmaco-
logic effects or abnormal behavior in the treated female rats
following dosing and during the observation period of 14 d
thereafter. All animals survived, gained normal body weight
and appeared active and healthy during the study. No gross
abnormalities or pathological alterations were noted for any
of the rats when necropsied at the conclusion of the 14-d
observation period. Based on these results and under the
conditions of this study, the acute oral LD
50
of FE is greater
than 5000 mg/kg of body weight in female rats.
Repeated dose 28-d sub-chronic toxicity study
All animals survived throughout the treatment period of 28 d.
No significant changes in the body weights, feed and water
intake were observed among any groups. There was no
abnormality found in any animal during ophthalmic examin-
ation before and after treatment except one. The reddening
was found only in one eye of one animal after low-dose
(250 mg FE/kg body weight/day) treatment. This was not a
dose-dependent effect and not found in other animals. So, this
was not treatment related. No statistically significant differ-
ence was observed in the motor activity scores for any dose
group when compared with the control animals. No changes
were observed among the parameters of urine analysis in any
of the groups when compared with the control groups.
The absolute organ weights of ovaries from mid- (500 mg/
kg body weight) and high-(1000 mg/kg body weight) dose
groups of female rats and spleen from high (1000 mg/kg body
weight) dose in male and female rats were found to be slightly
high, but not statistically significant when compared with
respective control groups. The relative organ weight of
ovaries does not show any significant difference.
The relative organ weights (organ to body weight ratio)
of spleen in mid- (500 mg/kg body weight) and high-
(1000 mg/kg body weight) dose groups in male rats were
found to be marginally high, but not statistically significant.
However, no microscopic abnormities were observed for
spleen and ovaries in histology evaluations, so the changes in
the organ weight have no toxicological relevance.
Hematology
In hematology, no statistical significant changes were seen in
any group of male rats when compared with the control
groups. However, marginal changes were observed in hem-
atocrit of low-dose group and eosinophils in mid- and high-
dose groups of female rats as compared to the control groups.
These changes were not considered to be toxicologically
relevant since effect also observed in the absence of clear
dose-related distribution, and also not present in male rats.
Clinical biochemistry
Among the various clinical biochemistry parameters esti-
mated, marginally high differences were only observed in the
cholesterol of high-dose group and creatinine in mid- and
high-dose groups of male rats as compared to control. In
female rats, albumin, ALT and potassium levels were
marginally higher in high-dose groups, but not statistically
significant, as compared to the control animals. The changes
observed in cholesterol of high-dose group in male rats and
albumin, ALT and potassium in high-dose group in female
rats were not considered to be toxicologically relevant since
these also occurred in the absence of clear dose-related
distribution. Furthermore, the changes in creatinine in mid-
and high-dose groups were minor and not significant in nature
and not found in female rats.
Gross pathological changes
At terminally sacrificed rats, the incidence of necropsy
findings in lungs like diffused red patches, congestion and
hemorrhage in liver were observed in few animals, but these
changes also occurred in control animals. These changes
observed in the organs might be because of adaptive
metabolic and physiological changes, anoxic/hypoxic condi-
tions during anesthesia and terminal sacrifice of the animals,
which affect the normal structure of the organs and were
considered to be dose-independent. So, these findings were
considered to be of no toxicological significance.
Microscopic histopathological findings
No remarkable changes were observed in control as well as in
high dose-group animals. However, few microscopic findings
observed in control animals including minimal focal collec-
tion of leucocytes in lobule and portal tract in liver and
increase in kupffer cells, hyperplasia of bronchial lymphoid
tissue and minimal focal increase in bronchi. In high-dose
group animals, hyperplasia of bronchial lymphoid tissue and
minimal focal infiltration of lymphocytes in lobule of liver
were observed in few animals. These findings were con-
sidered to be within the range of normal background lesions,
which may be seen in rats of this strain and during the aging
process of this study and were considered incidental in nature
with CO
2
inhalation and terminal changes at sacrifice,
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Toxicology Mechanisms and Methods Downloaded from informahealthcare.com by University of Georgia on 09/19/14
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reflecting the usual individual variability. As high-dose group
animals did not exhibit any toxic lesions on histological
examinations when compared with their respective control
group, hence further histological investigation was not
extended to the two other lower dose treatment groups.
Mutagenicity test: Ames’ bacterial reverse mutation
assay
No toxic effects of FE were noted in any of the five tester
strains used up to the highest dose group evaluated (with and
without metabolic activation). No biologically relevant
increases in revertant colony numbers of any of the five
tester strains were observed following treatment with FE at
any concentration level, in neither the presence nor absence of
metabolic activation. Therefore, FE did not cause gene
mutations by base pair changes or frameshifts in the
genome of the tester strains used, indicating that FE is non-
mutagenic. Also, no evidence of cytotoxicity was observed
in the form of reduction in the bacterial background lawn.
Finally, it is concluded that FE is non-mutagenic under the
conditions described for S. typhimurium Reverse Mutation
Assay (Ames test).
Anti-diabetic efficacy of FE in type 2 diabetic rat
model
Feeding of HFD for initially 2 weeks significantly (p50.05)
elevated body weight, PTG and PTC levels compared to
control group. The single dose of STZ (35 mg/kg, i.p.)-
induced diabetes (PGL 300 mg/dl) in almost all HFD-fed
animals with significant increase in 6-h fasting PGL, PTG
and plasma cholesterol levels as compared to the control
animals. However, no significant differences of plasma
insulin levels were observed between diabetic and normal
animals.
Chronic administration of FE (150 and 450 mg/kg body
weight) did not have significant influence on body weight
of animals, whereas glipizide treatment led to increase in
body weight as compared to vehicle-treated respective
control animals. No significant influence on feed intake
was observed in both FE- and glipizide-treated rats, while
glipizide-treated rats alone exhibited significant reduction
in water intake as compared to the corresponding
control animals.
FE (150 mg/kg body weight, p.o.) lowered PTG by
approximately 22.1%, 24.6% and 29.1% in STZ-induced
diabetic rats at 10-, 20- and 30- d of treatment, respectively,
while under these same conditions approximately 8.7%, 1.0%
and 12.4% reductions were observed at 10-, 20- and 30- d of
treatment with FE (450 mg/kg body weight, p.o.), respectively
(Table 1). Glipizide (5 mg/kg, p.o.) reduced PTG by 57.4%,
46.2% and 39.4% in diabetic rats at 10-, 20- and 30- d of
treatment, respectively (Table 1). FE (150 and 450 mg/kg
body weight) did not reduce PTC, while glipizide (5 mg/kg,
p.o.) reduced PTC by 23.9%, 37.1% and 19.7% in diabetic rats
at 10-, 20- and 30- d of treatment, respectively (Table 2). No
significant effect on 6-h fasting PI levels was observed upon
treatment with any of the treatment group as compared to the
corresponding control group. The chronic administration of
FE (150 and 450 mg/kg body weight) did not significantly
reduce PGLs in T2D rats as compared to the control animals
(Table 3).
Table 2. Effect of chronic administration on plasma total cholesterol levels (PTC) in streptozotocin-induced T2D rats.
PTC (mg/dl) at different intervals (% Reduction)
Groups Day 0 Day 10 Day 20 Day 30
Control 40.9 ± 2.0 38.8 ± 3.3 46.5 ± 3.2 50.4 ± 3.7
Diabetic control 168.5 ± 16.2* 191.6 ± 20.5* 198.6 ± 27.3* 181.4 ± 15.0*
Fenfuro (150 mg/kg) p.o. 161.2 ± 17.9* 212.1 ± 27.5* 164.7 ± 28.7* 155.4 ± 13.4*
Fenfuro (450 mg/kg) p.o. 167.1 ± 15.7* 207.4 ± 27.1* 176.0 ± 31.8* 174.4 ± 23.8*
Glipizide (5 mg/kg) p.o. 167.5 ± 17.5* 127.0 ± 12.6y(23.9%) 105.6 ± 13.4* (37.1%) 134.9 ± 16.6* (19.74%)
Fenfuro (150 and 450 mg/kg p.o.) and glipizide (5 mg/kg p.o.) were orally administered once daily for 20 consecutive days
and continued twice daily for the next 10 d, respectively, in streptozotocin-induced type 2 diabetic rats. Values are mean ± SEM
of 6–12 animals per group.
*p50.05 compared to the control group; yp50.05 compared to the diabetic control group.
Table 1. Effect of chronic administration on plasma triglyceride levels (PTG) in streptozotocin-induced T2D diabetic rats.
Groups Plasma triglyceride (mg/dl) at different intervals (% Reduction)
Day 0 Day 10 Day 20 Day 30
Control 55.6 ± 2.2 67.5 ± 7.4 65.6 ± 17.1 65.6 ± 7.2
Diabetic control 201.33 ± 22.5* 197.3 ± 21.2* 177.3 ± 19.1* 190.4 ± 21.9*
Fenfuro (150 mg/kg) p.o. 199.3 ± 15.8* 155.3 ± 14.2*y(22.1%) 150.0 ± 18.3*y(24.6%) 141.0 ± 23.0*y(29.1%)
Fenfuro (450 mg/kg) p.o. 185.0 ± 19.5* 161.6 ± 26.1*y(8.7%) 183.3 ± 29.4* (1.0%) 162.8 ± 18.0*y(12.4%)
Glipizide (5 mg/kg) p.o. 188.1 ± 9.1* 80.6 ± 10.4y(57.4%) 101.3 ± 8.4*y(46.2%) 114.7 ± 9.8y(39.4%)
Fenfuro (150 and 450 mg/kg p.o.) and glipizide (5 mg/kg p.o.) were orally administered once daily for 20 consecutive days and
continued twice daily for the next 10d, respectively, in streptozotocin-induced T2D rats. Values are mean ± SEM of 6–12
animals per group.
*p50.05 compared to the control group. yp50.05 compared to the diabetic control group.
500 A. Swaroop et al. Toxicol Mech Methods, 2014; 24(7): 495–503
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Effect of combination therapy (FE plus metformin) on
plasma glucose level
FE (150- and 450- mg/kg body weight, p.o.) extended for 10 d
did not significantly reduce plasma glucose level (PGL) in
diabetic rats, whereas metformin (100 mg/kg p.o.)-treated
animals exhibited insignificant reduction of PGL (Table 3).
Interestingly, oral administration of FE (150 mg/kg body
weight) in conjunction with metformin (100 mg/kg body
weight) twice daily for 10d exhibited marked and significant
reduction of 20.7% in PGL, while a combination of glipizide
and metformin significantly reduced PGL by 18% as
compared to the corresponding control group (Table 3). FE
(450 mg/kg body weight) in conjunction with metformin
exhibited a significant reduction of 14.3% (Table 3). These
results demonstrated a synergism between metformin and FE
at a 150 mg/kg body weight dose (Table 3). This potentiating
effect of FE on the anti-diabetic efficacy of metformin is
very interesting and warrants further investigation.
Another important point is that all the animals were found
healthy, active and normal as observed during quarantine and
acclimatization period of 7 d prior to the start of the study
and later throughout the study. No adverse events or mortality
was observed in any of the treated rats as compared to the
vehicle-treated rats.
Discussion
Fenugreek (Trigonella foenum-graecum, family Fabaceae) is
an annual plant cultivated worldwide as a semi-arid crop with
leaves consisting of three small obovate to oblong leaflets,
and cuboid-shaped, yellow-to-amber colored seeds (Adamska
& Lutomski, 1971; Singhal et al., 1982). Both fenugreek
leaves and seeds have long been used in Ayurvedic and
Chinese medicines in the treatment of diabetes. The leaves
and seeds are also regularly consumed as foods, vegetables
and spices in the Indian subcontinent, China and middle-
eastern countries (Stark & Madar, 1993). It is important to
mention that India is the largest producer of Fenugreek (http://
en.wikipedia.org/wiki/Fenugreek). For culinary purpose,
fenugreek seeds are often roasted to reduce bitterness and
enhance flavor. Fresh fenugreek leaves are integral constituent
in some Indian curries, while sprouted seeds and microgreens
are used in salads (Bordia et al., 1997).
Active constituents of fenugreek seeds include soluble
dietary fiber, protein, vitamin C, niacin, potassium, 4-hydro-
xyisoleucine, lysine and selected amino acids, L-tryptophan,
and selected steroidal saponins including diosgenin, yamo-
genin, tigogenin and neotigogenin, which have been demon-
strated to inhibit both cholesterol absorption in the intestine
and cholesterol production by the liver (Gupta et al., 1986;
Valette et al., 1984). It has been reported that the fenugreek
seeds contain a gel-like soluble fiber which combines with
bile acids and lowers the TG and LDL levels. These amino
acids are useful in boosting insulin sensitization and glycogen
synthesis. Hypoglycemic activity of fenugreek has mainly
been attributed to dietary fibers and saponins. Especially
the dietary fiber may significantly contribute to fenugreek’s
activity in lowering blood glucose and cholesterol
(Madar et al., 1988).
Numerous therapeutic efficacy and medicinal properties of
fenugreek on metabolic disorders have been demonstrated.
In recent years, fenugreek is being used in the Western world
as a medicinal herb or as a spice (Bordia et al., 1997; Elujoba
& Hardman, 1987). Fenugreek has been reported to reduce
serum cholesterol level and attenuate blood glucose level and
improves lipid metabolism. It has also been reported that
fenugreek reduces lipid level in plasma and liver leading to
the improvement of insulin sensitivity in rats with metabolic
disorders (Madar et al., 1988; Prasanna, 2000).
Animal studies suggest that fenugreek may also contain a
constituent which stimulates insulin production or sensitiza-
tion (Gupta et al., 1986; Sauvaire et al., 1991). Earlier studies
in animals demonstrated that fenugreek seed extract has the
potential to slow the enzymatic digestion of carbohydrates,
reduced gastrointestinal absorption of glucose and thus reduce
post-prandial glucose level (Elujoba & Hardman, 1987;
Sauvaire et al., 1991).
Multiple human trials have found that fenugreek may help
lower total cholesterol in people with moderate atheroscler-
osis or insulin- or non-insulin-dependent diabetes. One
human double-blind trial has demonstrated that defatted
fenugreek seeds may raise the beneficial HDL cholesterol
(Neelakantan et al., 2014; Sharma et al., 1991). Another
human study found that two different dosage of fenugreek of
25 or 50 g per day of defatted fenugreek seed powder
significantly lowers serum cholesterol after 20 d
Table 3. Effect of chronic administration of Fenfuro and Metformin, singly and in combination, on plasma glucose
levels (PGL) in streptozotocin-induced T2D rats.
Groups Plasma glucose level (PGL mg/dl) (% Reduction)
Control 104.3 ± 5.1 100.4 ± 5.2
Diabetic control 404.3 ± 22.8* 424.3 ± 35.1*
Metformin (100 mg/kg) p.o. 412.8 ± 10.0* 370.9 ± 6.1* (%)
Fenfuro (150 mg/kg) p.o. 405.7 ± 18.8* 372.5 ± 13.0* (8.1%)
Fenfuro (150 mg/kg) ± Metformin (100 mg/kg) p.o. 420.6 ± 14.9* 333.3 ± 15.6*y(39.4%)
Fenfuro (450 mg/kg) p.o. 409.4 ± 21.4* 420.1 ± 14.5* (2.6%)
Fenfuro (450 mg/kg) ± Metformin (100 mg/kg) p.o. 428.2 ± 13.9* 367.9 ± 23.8*y(14.3%)
Glipizide (5 mg/kg) p.o. 430.3 ± 19.5* 409.7 ± 27.4* (4.8%)
Glipizide (5 mg/kg) ± Metformin (100 mg/kg) p.o. 411.1 ± 17.0* 337.6 ± 1.9 (18.0%)
Fenfuro (150 and 450 mg/kg p.o.), metformin (100 mg/kg p.o.) and glipizide (5mg/kg p.o.) were orally administered
once daily for 20 consecutive days and continued twice daily for the next 10 d, respectively, in streptozotocin-
induced type 2 diabetic rats. Values are mean ± SEM of 6–12 animals per group.
*p50.05 compared to the control group; yp50.05 compared to the diabetic control group.
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(Prasanna, 2000). In a controlled trial, people suffering from
T2D taking 15 g of powdered fenugreek in meals reduced the
rise in blood glucose after the meal. Another similar
controlled trial found that taking 2.5 g of fenugreek twice a
day for 3 months reduced blood sugar level in people with
mild T2D (Sauvaire & Baccou, 1978; Sauvaire et al., 1991).
In another double-blind study with T2D subjects, 1 g per day
of an extract of fenugreek seeds for 2 months improved blood
sugar and insulin function. Fenugreek has been shown to
lower total and LDL cholesterol and triglyceride level in
people with high lipid level (Bordia et al., 1997; Neelakantan
et al., 2014). A small randomized trial demonstrated that
fenugreek treatment of 100 g per day lower elevated blood
glucose level, triglyceride and various lipids (Neelakantan
et al., 2014; Prasanna, 2000). Fenugreek treatment in humans
and animals demonstrated significant attenuation of glucose
tolerance and improvement in the glucose-induced insulin
response, suggesting a promising hypoglycemic effect of
fenugreek (Adamska & Lutomski, 1971; Neelakantan et al.,
2014; Prasanna, 2000).
The current study demonstrated the broad-spectrum safety
of FE in animals over the dose levels and routes of
administration tested. Acute oral toxicity did not reveal any
significant changes in multiple species for all examined
tissues. Based on the results, the oral LD
50
of FE was
concluded to be 45000 mg/kg in female rats. Repeated dose
28-d oral administration of FE to Sprague–Dawley rats at 0,
250, 500 and 1000 mg/kg/day caused no mortality, no
toxicologically relevant changes, hematological, histopatho-
logical or clinical abnormalities in the rats tested under the
conditions of the experiment. Extensive blood chemistry
analysis demonstrated broad spectrum safety. Ames’ Bacterial
Reverse Mutation Assay using five strains of S. typhimurium
(TA97A, TA98, TA100, TA102, TA1535, TA1537 and
TA102) was used to evaluate the mutagenic potential of FE
in the presence and absence of metabolic activation in the
L5178Y mouse lymphoma cell line. FE did not induce
mutagenic effects either with or without metabolic activation.
Taken together, these studies demonstrate the broad spectrum
safety of FE. FE (150 mg/kg p.o.) reduced the PTG levels in
T2D rats by 22%, 24.6% and 29% at 10-, 20- and 30-d
of treatment, respectively, while glipizide (5 mg/kg p.o.)
reduced the PTG levels by 57.4%, 46.2% and 39.4% at these
time points.
Furthermore, male Sprague–Dawley rats were given HFD
and STZ (35 mg/kg i.p.) to induce T2D, FE (150 and 450 mg/
kg p.o.) and glipizide (5 mg/kg p.o.) were administered over a
period of 30 d and fasting plasma TG, TC and insulin levels
were assessed. No effect was observed on body weight, feed
and water intake in T2D rats. FE (150 mg/kg p.o.) reduced the
PTG levels in T2D rats by 22%, 24.6% and 29% at 10-, 20-
and 30-d of treatment, respectively, while glipizide (5 mg/kg
p.o.) reduced the PTG levels by 57.4%, 46.2% and 39.4% at
these time points. This demonstrated that FE provided a
steady time-dependent improvement in the PTG levels. FE
(450 mg/kg) demonstrated significant hypoglycemic activity
(31.5%) as compared to insulin (48.2% with 1 U/kg i.p.). FE
(150 mg/kg p.o.) and metformin (100 mg/kg p.o.) combination
produced significant reduction (20.7%) on PGL in T2D rats.
No adverse effects were observed.
Currently, we are exploring the mechanistic insight to
understand the intricate pathophysiology. Human clinical
trials are in progress to assess the efficacy of FE at
different doses.
Acknowledgements
This study was presented in part in the 53rd Annual Meeting
of the Society of Toxicology in Phoenix, AZ, USA.
Declaration of interest
A.S., P.K., K.T., M.B. and D.B. are working in the company,
while Dr. Preuss is a professor at Georgetown University
Medical Center, Washington, DC. Dr. P.A. Marone is the
executive director of Eurofins/Product Safety Labs, Dayton,
NJ, and directed the safety studies.
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... As a result, the compound treatment significantly decreased TC, LDL, HDL, glucose-6-phosphate, sorbitol dehydrogenase, and aldose reductase while enhancing the level of the liver and muscle glycogen, hexokinase, glucokinase, pyruvate kinase, malic enzyme, glucose-6-phosphate dehydrogenase, SOD, and GPx and restored normal glucose level. Swaroop et al. (2014) assessed the antidiabetic effect of Trigonella seed extract on STZ induced T2D in male Sprague-Dawley rats. The rats were administrated with 150 mg/kg of the extract, noticed with 22%, 24.6%, and 29% reduction of plasma triglyceride at 10, 20 and 30 days of therapy, respectively. ...
... The extract also showed an excellent tolerability profile (Nathan et al., 2014). Swaroop et al. (2014) examined the antidiabetic effect of Trigonella seed extract on STZ induced T2D in male Sprague-Dawley rats. The extract showed a significant antidiabetic effect without any adverse effects. ...
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Trigonella foenum-graecum (fenugreek) belonging to the family Fabaceae, is widely used for both culinary as well as clinical purposes since antiquity. Folkloric medicines across globe particularly use this plant for boosting immunity and combating digestive and reproductive impairments. The plant is rich reservoir of different phytoconstituents attributed to their diverse pharmacological effects. Therefore, the present article is planned on its ethnomedicinal uses, botanical description, phytochemistry, pharmacology, toxicology, clinical efficacy, mechanism of action and nanoparticle synthesis. In all pharmacological studies, the dose, major bioactive, type of extract and possible outcomes is also discussed, to establish its specific role against a particular ailment. It was hypothesized that the nanoparticle synthesis will lead to the enhanced pharmacology. Results showed that ethnomedicinal data well supports the different pharmacological aspects of Trigonella formulations in different countries. Trigonelline (phytoestrogen) renders most of therapeutic potential of Trigonella. The reported therapeutics can also be accounted as the synergistic pharmacology of different bioactives. Nanoparticle synthesis significantly improves its pharmacological efficacy. Clinical studies well validated its antidiabetic and reproductive health improving efficacies. Though no serious toxic effects were observed with the use of this plant but further well-designed placebo trials are still needed to demonstrate its full therapeutic potential.
... Trigonella foenumgraecum is the fourth plant used by the HAC population. Several studies had demonstrated the hypoglycemic effects of fenugreek (Kumar et al., 2012;Swaroop et al., 2014), moreover, the results of the study of Sharma et al. (1990) had concluded that fenugreek grains were useful in the control of glycemia. Recent study, published in 2017, concluded that Rosmarinus officinalis is effective in the traditional treatment of diabetes mellitus (Hamidpour et al., 2017). ...
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Context: Diabetes mellitus is considered one of the scourges of the third millennium in the world for several years. Aims: To evaluate, identify and preserve the experiences gained over the centuries. It is about therapeutic use of medicinal plants exploited for the treatment of diabetes, and to clarify its toxicities, in order to sensitize the population of the High Atlas Central (HAC) of Morocco not only on the risk but also the benefit of the use of phytotherapy. Methods: The surveys ethnopharmacological were conducted 834 the interviewees, using semi-structured survey, by application of quantitative indices such as Consensus index (CI %), Use value (UVi), family UV (FUV), relative frequency of citation (RFC), plant part value (PPV), and informant consensus factor (ICF) to assess the exact value of medicinal plants (MP). Results: We found that 144 medicinal plants in 121 genera and 52 plant families were traditionally used to treat diabetes, of which seven species were endemic to the study area, and 32 were being cited first ever to treating diabetes. In addition, we collected toxicological information on 99 antidiabetic plants, of which 41 species showed no toxic activity and 43 were toxic and sometimes lethal in high doses. The Ranunculaceae family showed the highest use value (FUV =0.139). Leaves were the most used plant parts (PPV =0.282) and infusion was the dominant method of preparation. The frequently used species were Olea europaea L. (UVi =0.172), Salvia officinalis L. (UVi =0.156) and Euphorbia resinifera Berg (UVi =0.150). Conclusions: These results are a rich source of information. They contribute to the knowledge of the antidiabetic medicinal flora of our study area, and to the preservation of the local popular know-how of the word that tends to disappear. They may also represent a database that consists of purifying and identifying the characterization of active compounds of herbal extracts with antidiabetic activity.
... Trigonella foenumgraecum is the fourth plant used by the HAC population. Several studies had demonstrated the hypoglycemic effects of fenugreek (Kumar et al., 2012;Swaroop et al., 2014), moreover, the results of the study of Sharma et al. (1990) had concluded that fenugreek grains were useful in the control of glycemia. Recent study, published in 2017, concluded that Rosmarinus officinalis is effective in the traditional treatment of diabetes mellitus (Hamidpour et al., 2017). ...
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Resumen Context: Diabetes mellitus is considered one of the scourges of the third millennium in the world for several years. Aims: To evaluate, identify and preserve the experiences gained over the centuries. It is about the therapeutic use of medicinal plants exploited for the treatment of diabetes, and to clarify its toxicities, in order to sensitize the population of the High Atlas Central (HAC) of Morocco not only on the risk but also the benefit of the use of phytotherapy. Methods: The surveys ethnopharmacological were conducted 834 the interviewees, using semi-structured survey, by application of quantitative indices such as Consensus index (CI %), Use value (UVi), family UV (FUV), relative frequency of citation (RFC), plant part value (PPV), and informant consensus factor (ICF) to assess the exact value of medicinal plants (MP). Results: We found that 144 medicinal plants in 121 genera and 52 plant families were traditionally used to treat diabetes, of which seven species were endemic to the study area, and 32 were being cited first ever to treating diabetes. In addition, we collected toxicological information on 99 antidiabetic plants, of which 41 species showed no toxic activity and 43 were toxic and sometimes lethal in high doses. The Ranunculaceae family showed the highest use value (FUV =0.139). Leaves were the most used plant parts (PPV =0.282) and infusion was the dominant method of preparation. The frequently used species were Olea europaea L. (UVi =0.172), Salvia officinalis L. (UVi =0.156), and Euphorbia resinifera Berg (UVi =0.150). Conclusions: These results are a rich source of information. They contribute to the knowledge of the antidiabetic medicinal flora of our study area, and to the preservation of the local popular know-how of the word that tends to disappear. They may also represent a database that consists of purifying and identifying the characterization of active compounds of herbal extracts with antidiabetic activity. Contexto: La diabetes mellitus se considera uno de los flagelos del tercer milenio en el mundo desde hace varios años. Objetivos: Evaluar, identificar y preservar las experiencias adquiridas a lo largo de los siglos. Se trata del uso terapéutico de las plantas medicinales explotadas para el tratamiento de la diabetes, y de aclarar sus toxicidades, a fin de sensibilizar a la población del Alto Atlas Central (HAC) de Marruecos no sólo sobre el riesgo sino también sobre el beneficio del uso de la fitoterapia. Métodos: Las encuestas etnofarmacológicas se realizaron 834 a los entrevistados, utilizando una encuesta semiestructurada, mediante la aplicación de índices cuantitativos como el índice de consenso (CI %), el valor de uso (UVi), la familia UV (FUV), la frecuencia relativa de citación (RFC), el valor de la parte de la planta (PPV) y el factor de consenso de los informantes (ICF) para evaluar el valor exacto de las plantas medicinales (MP). Resultados: Encontramos que 144 plantas medicinales en 121 géneros y 52 familias de plantas se usaban tradicionalmente para tratar la diabetes, de las cuales siete especies eran endémicas del área de estudio y 32 se citaban por primera vez para tratar la diabetes. Además, recopilamos información toxicológica sobre 99 plantas antidiabéticas, de las cuales 41 especies no mostraron actividad tóxica y 43 eran tóxicas y, a veces, letales en dosis altas. La familia Ranunculaceae mostró el valor de uso más alto (FUV = 0,139). Las hojas eran las partes de las plantas más utilizadas (VPP = 0,282) y la infusión el método de preparación predominante. Las especies más utilizadas fueron Olea europaea L. (UVi = 0,172), Salvia officinalis L. (UVi = 0,156) y Euphorbia resinifera Berg (UVi = 0,150). Conclusiones: Estos resultados son una rica fuente de información. Contribuyen al conocimiento de la flora medicinal antidiabética de la zona de estudio, y a la preservación del saber popular local de la palabra que tiende a desaparecer. También pueden representar una base de datos para depurar e identificar la caracterización de compuestos activos de extractos de hierbas con actividad antidiabética.
... Plants having alkaloids, terpenoids, glycosides and flavonoids exhibit antioxidant properties which is responsible for driving their hypoglycaemic actions, by regenerating the damaged beta pancreatic cells in addition to saponin induced inhibition of Na-Glucose co transporter-1 in intestine. [17][18][19][20] Phytochemical screening of both Achillea and Caralluma confirmed the presence of flavonoids, alkaloids and saponin, suggesting similar modes of hypoglycaemic action. To determine the exact underlying mechanisms, a detailed phytochemical and pharmacological profiles of these plants are therefore necessary to establish. ...
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Diabetes mellitus affects approximately 20 million people in the US, or nearly 7% of the population. It is expected to increase by 70% within the next 25 years and numerous epidemiologic studies have demonstrated that type 2 diabetes increases the risk of cardiovascular morbidity and mortality. It is estimated to cost over $92 BILLION in health care costs and lost productivity. The increased risk is due to the detrimental vascular effects of prolonged exposure to a hyperglycemic, oxidant rich environment yielding associated cardiovascular risk factors: atherosclerosis, hypertension and clotting abnormalities. Hypertension and dyslipidemia in diabetic patients produces substantial decreases in cardiovascular and micro vascular diseases. NUTRITIONAL AND THERAPEUTIC INTERVENTION OF DIABETES AND METABOLIC SYNDROME provides an overview of the current epidemic, outlines the consequences of this crisis and lays out strategies to forestall and prevent diabetes, obesity and other intricate issues of metabolic syndrome. The contributing experts from around the world give this book relevant and up-to-date global approaches to the critical consequences of metabolic syndrome and make it an important reference for those working with the treatment, evaluation or public health planning for the effects of metabolic syndrome and diabetes. Scientific discussion of the epidemiology and pathophysiology of the relationship between diabetes and metabolic syndrome Includes coverage of Pre-diabetes conditions plus both Type I and Type II Diabetes Presents both prevention and treatment options.
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