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Research Article
Preclinical Safety of the Root Extract of Polygala tenuifolia
Willdenow in Sprague-Dawley Rats and Beagle Dogs
Ki Young Shin,1,2 Beom Young Won,1Hyun Jee Ha,1Yeo Sang Yun,1and Hyung Gun Lee1
1Research & Development Center, Braintropia Co. Ltd., Anyang-si, Gyeonggi-do 431-716, Republic of Korea
2Department of Microbiology, College of Natural Science, Dankook University, Cheonan-si, Chungnam 330-714, Republic of Korea
Correspondence should be addressed to Ki Young Shin; newsky@braintropia.com
Received July ; Revised October ; Accepted October ; Published November
Academic Editor: Khalid Rahman
Copyright © Ki Young Shin et al. is is an open access article distributed under the Creative Commons Attribution License,
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
e root of Polygala tenuifolia Willdenow has been used for the treatment of insomnia, depression, and amnesia. However, the
toxicological properties of the herb have been overlooked, because it has been used for a long time for various purposes. In this
study, we evaluated the preclinical safety of the root extract in rats and beagle dogs. First, the acute oral toxicity was tested in
both rats and dogs. In the rats, only one female of g/kg died, but no treatment-related death or clinical and gross ndings were
observed aer the administration. No toxicological changes or mortalities related to the test substance were also observed aer
the administration in the dogs. Although vomiting, discoloration, or hemorrhage was found in some dogs, there were no serious
abnormalities. Second, the subchronic toxicity was investigated in the rats. Two animals were found dead in the female group of
, mg/kg/day, but there were no abnormal ndings associated with the test substance. ere also were no adverse eects on the
clinical signs, body weight, and hematological and biochemical ndings. erefore, our results showed that the acute or subchronic
toxicity of the root extract of Polygala tenuifolia might not be toxic to rats and dogs.
1. Introduction
Traditional oriental herbal prescriptions have regained popu-
larity over the past decade. ey are used widely for the treat-
ment and prevention of various diseases [,], because their
long history of clinical application and natural origin seem to
guarantee that the prescriptions will be eective and nontoxic
[].Eachplantusedinsuchherbalprescriptionscanprovide
opportunities for the development of herbal food products,
dietary supplements, and functional foods []. However,
the toxicological aspects of medicinal herbs or naturally
occurring functional foods have been neglected due to their
long history of use. It has been demonstrated that medicinal
plants may have undesirable properties [], because some
individuals taking herbal medicines have reported certain
side eects. erefore, the use of any plant for medicinal
purposes by no means guarantees the safety of such a plant.
is fact raises concerns about the potential toxic eects
resulting from short-term and long-term use of such medic-
inal plants. Data from acute and subchronic toxicity studies
on medicinal plants or naturally occurring functional foods
should be obtained to increase condence in the safety of
their use, particularly in the development of pharmaceuticals
[]. Polygala tenuifolia Willdenow is a perennial herbaceous
plant distributed widely in China and Korea. In Asia, it is a
well-known traditional medicine for the treatment of phlegm
and detumescence. Traditional medicines are prepared from
the roots of P. t e n u i f o l i a because of its expectorant, tonic,
tranquilizing, and antipsychotic properties [–]. In particu-
lar, the herb is used against insomnia, neurasthenia, amnesia,
depression, anxiety-related palpitations, restlessness, disori-
entation, dementia, and memory failure [,]. Previously,
we reported that the root extract of P. t e n u i f o l i a was shown
to enhance memory and cognitive function in two animal
modelsandtwohumanmodels[–]. e extract has been
shown to contain C-glycosides, triterpene saponins, sucrose
esters, and oligosaccharide esters [,]. It also contains
various substances, such as tenuigenin, tenuifolin, DISS (,-
disinapoyl sucrose), and TMCA (,,-trimethoxycinnamic
acid), which have been shown to have proliferative and
protective eects on hippocampal neurons [,]. Several
Hindawi Publishing Corporation
Evidence-Based Complementary and Alternative Medicine
Volume 2014, Article ID 570134, 15 pages
http://dx.doi.org/10.1155/2014/570134
Evidence-Based Complementary and Alternative Medicine
researchers have demonstrated the eects of P. t e n u i f o l i a ,but
information on its safety is lacking. erefore, systematic
evaluation of the safety of the root extract of this herb
is necessary for the development of new foods or drugs.
In this study, an alcohol extract from the dried root of P.
tenuifolia was prepared, and its safety was evaluated using
an acute oral toxicity test and a subchronic oral toxicity test
in Sprague-Dawley rats and beagle dogs in accordance with
two standards: the Korea Food and Drug Administration
Notication numbers - and -.
2. Materials and Methods
2.1.PreparationoftheRootExtractofPolygalatenuifolia. e
dried root extract of Polygala tenuifolia ( g) was reuxed
with % ethanol for hr in a boiling water bath. is
procedure was repeated twice and the ethanol solution was
concentrated under a vacuum. e resulting concentrated
ethanol fraction ( g) of the plant root was used for this
study [].
2.2. Experimental Animals and Animal Husbandry. Six-
week-old specic pathogen-free Sprague-Dawley (HSD; SD)
rats were used. Six animals of each sex for the acute oral toxi-
citystudyandanimalsofeachsexforthesubchronicoral
toxicity study were received from Koatech Co. (Pyeongtaek-
si, Gyeonggi-do, Korea), Ltd. For the acute oral toxicity study,
sixbeagledogsofeachsexatmonthsofagewereobtained
fromtheHanlim Laboratory Animals Company (Hwaseong-
si, Gyeonggi-do, Korea). e animal room was maintained at
atemperatureof±∘C, with a relative humidity of ±
%, air ventilation of – times/h, and ambient light (–
Lux) controlled to produce a h light/dark cycle. Ani-
mals were given irradiation-sterilized pellet feed and ground
water disinfected by an ultraviolet sterilizer and ultraltered
ad libitum. e dogs were reared in stainless steel cages (
× × mm), and identication cards showing their
study number and animal number were attached to the cages.
2.3. Study Design Overview. is study was conducted in
compliance with the Good Laboratory Practice (GLP) and
Test Guidelines of the Organization for Economic Coopera-
tion and Development [,] and the Korea Food and Drug
Administration [,]attheGLPInstituteapprovedbythe
KFDA (Korea Food and Drug Administration). e study
protocol was approved by the Institutional Animal Care and
Use Committee of the institute, which is accredited by the
Association for Assessment and Accreditation of Laboratory
Animal Care International, .
2.3.1. Acute Oral Toxicity Study in Rats. Aer a -day quaran-
tine and acclimatization period, ten healthy animals of each
gender at weeks of age were used. e study included two
groups: a control group given no treatment and an exper-
imental group given , mg/kg, based on a preliminary
study (data not shown). Each group consisted of ve rats
of each sex, and body weight ranges at dosing were .–
. g for males and .–. g for females. e test
substance was administered orally to each group. ereaer,
clinical signs and body weight were observed throughout the
-day experimental period, and gross ndings were observed
on the scheduled necropsy day. is study was carried
out in accordance with the standard operating procedures
of SNUH-GLP (Non-Clinical Research Center, Biomedical
Research Institute, Seoul National University Hospital).
2.3.2. Acute Oral Toxicity Study in Beagle Dogs. Six male
and six female dogs were divided into three groups of four
animals; then, two dogs of each gender were allocated. e
study included three groups: a control group given no treat-
ment and experimental groups exposed to , mg/kg and
, mg/kg. e body weight ranges at dosing were .–
.kgformalesand.–.kgforfemales.etest
substance was administered orally to each group. ereaer,
clinical signs, mortality, and body weight were observed
throughout the -day experimental period, and gross nd-
ings were observed on the scheduled necropsy day. is study
was carried out in accordance with the standard operating
procedures of Preclinical Research Center, ChemOn (Yongin,
Gyeonggi-do, Korea) Inc.
2.3.3. Subchronic Oral Toxicity Study in Rats. Aer a -day
quarantine and acclimatization period, forty healthy animals
ofeachgenderatweeksofagewereused.Adoseof
, mg/kg/day was set as the high dose considering the
results of a preliminary -week subchronic oral toxicity study
(data not shown) and the characteristics of the test substance.
Doses of and mg/kg/day were set as the middle and
low doses, respectively, using a common ratio of .. e
vehicle-treated group was administrated with sterile water
for injection only. Each group consisted of rats of each
sex, and rats of each sex and group were killed aer a -
day treatment period. Body weight ranges at the beginning
of dosing were .–. g for males and .–. g
for females. e test substance was administered repeatedly
by the oral route to each group for days. e following
items were examined during the experimental period: clinical
signs, body weight, food and water consumption, ophthalmic
examination, urinalysis, hematology, blood clotting time,
serum biochemistry, necropsy ndings, organ weight, and
histopathology. is study was carried out in accordance with
the standard operating procedures of Preclinical Research
Center, ChemOn Inc.
2.4. Observation and Examination Items of the Acute Oral
Toxicity Study in Rat s
2.4.1. Clinical Signs. In the rat studies, clinical signs and
mortality were observed continuously for the rst h aer
administration and every hour for h. Each animal was
observed daily throughout the entire -day experimental
period. Abnormal type and severity of signs, as well as
the observation day and time, were recorded. e day of
administration was set as day .
2.4.2. Body Weight. Each rat was measured on days , , ,
and using an electronic balance (Sartorius Co., Gottingen,
Lower Saxony, Germany).
Evidence-Based Complementary and Alternative Medicine
2.4.3. Necropsy. On the scheduled necropsy day (day ), all
surviving animals were euthanized by exsanguination from
the abdominal aorta and abdominal vena cava under a CO2
gas overdose and examined grossly for abnormalities of the
body surface and internal organs.
2.5. Observation and Examination Items of the Acute Oral
Toxicity Study in Dogs
2.5.1. Clinical Signs. In the dog studies, the clinical signs
and mortalities were checked every hour for hours aer
administration. Each animal was observed daily throughout
the entire -day experimental period. Abnormal type and
severity of signs, as well as the observation day and time, were
recorded. e day of administration was set as day .
2.5.2. Body Weight. All dogs were individually weighed
beforeadministrationandondays,,,andaerthe
administration.
2.5.3. Necropsy. In the dog studies, the animals were anes-
thetized with ketamine (Yuhan Yanghaeng) and rompun
(Bayer Korea) and euthanized by exsanguinations from the
axillary artery and vein. All organs in the thoracic and
abdominal cavities were observed grossly and the gross
ndings were recorded.
2.6. Observation and Examination Items of the Subchronic
Oral Toxicity Study in Rats
2.6.1. Clinical Signs. All animals were observed daily for
clinical signs and mortality. e type, date of occurrence, and
the severity of the signs were recorded individually. e initial
day of administration was set as day .
2.6.2. Body Weight. Animals were weighed on the initial day
of administration, once per week during the experimental
period, and on the day of the necropsy. e body weight at
the necropsy was measured aer an overnight fast.
2.6.3. Food and Water Consumption. Food and water con-
sumption were measured on the initial day of administration
and then once per week during the experimental period. e
amounts of food and water were measured before they were
suppliedtoeachcage,andthefoodandwaterremainingthe
next day were measured to calculate the dierence, which was
regarded as daily food and water consumption (g/rat/day).
2.6.4. Ophthalmic Examination. In the last week of obser-
vation, a mydriatic (Ocuhomapin, Lot number , Samil
PharmCo.,Korea)wasdroppedintobotheyesofeachanimal
to facilitate mydriasis aer observing the external appearance
of the eyes of ve males and ve females per group. e
anterior parts of the eyes, the optic media, and the ocular
fundus were then observed with an ophthalmoscope and a
fundus camera (Genesis, Kowa Co., Tokyo, Japan). No abnor-
mal signs were observed during the ophthalmic examination.
erefore, no eye photographs or further examinations were
performed.
2.6.5. Urinalysis. Five males and ve females per group were
housed in metabolic cages for urine collection in the last week
of obser vation, and fresh urine samples (about mL) were
collectedforhandusedforurinalysisandurinesediment
tests. Additionally, the total urine volume was measured
from urine samples collected for h. Test strips (Multistix
SG, Siemens, Washington, DC, USA) were dipped in
about . mL of urine and the specic gravity, pH, protein,
glucose, ketone bodies, occult blood, bilirubin, urobilinogen,
and nitrite levels were analyzed with an automatic analyzer
(CliniTek , Siemens, Ames Division, Miles Laboratory,
USA). Urine color was observed with the naked eye, and the
result was input into the automatic analyzer. Approximately
. mL of urine was centrifuged (Hanil MF, Seoul, Korea)
for min. e sediment was stained using the Sternheimer-
Malbin method (Sternheimer and Malbin, ) and red
blood cells (RBCs), white blood cells (WBCs), epithelial cells,
and casts were observed under a microscope.
2.6.6. Hematology. Approximately mL of the blood taken
during the necropsy was placed into a CBC bottle (Vacutainer
mL, Becton Dickinson, Franklin Lakes, NJ, USA) contain-
ing EDTA-K anticoagulant. e following parameters were
measured with a Coulter counter (ADVIA , Siemens,
Ames Division, Miles Laboratory, USA): white blood cell
count, red blood cell count, hemoglobin, hematocrit, mean
corpuscular volume, mean corpuscular hemoglobin, mean
corpuscular hemoglobin concentration, red cell distribu-
tion width, hemoglobin distribution width, platelets, mean
platelet volume, and WBC dierential count (neutrophils,
lymphocytes, monocytes, eosinophils, basophils, and large
unstained cells). . mL of the blood taken during the
necropsy was dispensed into a microtube containing . mL
.% sodium citrate, and plasma was obtained from cen-
trifugation (Eppendorf, Hamburg, Germany) for min to
assess the blood clotting time. e prothrombin time and
activated partial thromboplastin time were measured in sec-
onds from plasma using the nephelometric analysis method
(Woziwodzki, ) with a coagulation time analyzer (ACL
, Instrumentation Laboratory, Bedford, MA, USA).
2.6.7. Serum Biochemistry. More than mL of the blood taken
duringthenecropsywasaddedtoamLVacutainertube
(IMPROBE, Germany) containing a clot activator. e blood
was coagulated by maintaining it at room temperature for
– min and was then centrifuged (, rpm, , RCF,
MF, Hanil, Korea) for min. e following parameters
were measured with a serum biochemistry analyzer (AU,
Olympus, Tokyo, Japan): aspartate aminotransferase, alanine
aminotransferase, alkaline phosphatase, creatine phosphoki-
nase, total bilirubin, glucose, total cholesterol, triglycerides,
total protein, albumin, albumin/globulin ratio, blood urea
nitrogen, creatinine, inorganic phosphorus, and calcium
ions. e electrolytes were measured with the electrolyte
autoanalyzer ( Na, K, Cl Analyzer, Ciba-Corning, USA).
2.6.8. Necropsy and Organ Weight. Before the scheduled
necropsy, all surviving animals were fasted overnight (for
Evidence-Based Complementary and Alternative Medicine
– h) and euthanized with isourane (Ifran liquid, Hana
Pharm. Co., Korea) inhalation on the day of the necropsy.
Aer anesthesia was conrmed, blood was taken from the
posterior vena cava for hematology and serum biochemistry
analyses. e abdominal aorta and posterior vena cava were
cuttoeuthanizetheanimals.Allorgansofthebodysurface,
subcutis, head, and all internal organs of the abdominal and
thoracic cavities were observed grossly. Next, the pituitary
gland, ovaries, uterus, adrenal glands, thymus, prostate gland,
testes, epididymides, spleen, kidneys, heart, lung, brain, and
liver were removed and weighed with an electronic balance
(BPS, Sartorius Co., Gottingen, Lower Saxony, Germany),
and all paired organs were measured separately. e absolute
organ weights were converted to relative organ weights based
on the organ-to-fasted body weight ratios.
2.6.9. Histopathology. Microscopic examinations were per-
formedonthepreservedorgansandtissuestakenfrom
all animals in the vehicle-treated and high dose-treated
groups. All gross lesions as dened by the study pathologist
were also included in the examination. e eyes and the
optic nerves were preserved in Davidson’s xative, and the
testes and epididymides were preserved in Bouin’s xative.
e following organs and tissues were xed in % neutral
buered formalin solution: the thymus, spleen, pancreas,
stomach, duodenum, jejunum, ileum, cecum, colon, rectum,
mesenteric lymph nodes, mandibular lymph nodes, salivary
gland, thyroid gland (including parathyroid gland), Harde-
rian gland, heart, lung, kidneys, adrenal glands, liver, aorta,
brain, pituitary gland, tongue, trachea, esophagus, sternum,
thoracic spinal cord, femorotibial joint, peripheral nerve
(sciatic), skeletal muscle (femoral), prostate gland, seminal
vesicles, ovaries, uterus, vagina, urinary bladder, and skin
(including mammary gland).
2.7. Statistical Analysis. In the rat studies, data are presented
as mean ±standard deviation. Body weight, food and water
consumption, total volume of urine, hematology and serum
biochemistry,andabsoluteandrelativeorganweightswere
assumedtobenormallydistributedandwereanalyzedbya
one-way analysis of variance (ANOVA) []. e assumption
of homogeneity was tested using Levene’s test []. If the
overall ANOVA was signicant and the assumption of homo-
geneity of variance was met, Duncan’s multiple-range test
was used as a post hoc test to identify signicantly dierent
groups from the vehicle control group []. Schee’s test was
used if the sample size was unequal between the groups [].
If the assumption of homogeneity of variance was not met,
Dunnett’s T test was used as the post hoc test []. Student’s
𝑡-test was used to analyze the dierences in means between
the two independent groups. e urinalysis data were rank-
transformed and analyzed by the nonparametric Kruskal-
Wall i s 𝐻-test []. If a statistically signicant dierence was
observed between groups, the Mann-Whitney 𝑈test was used
to identify the groups that were signicantly dierent from
the vehicle control group []. SPSS .K was used for all
statistical analyses (Chicago, IL, USA). A 𝑃 < 0.05 was
considered signicant.
In the dog studies,data are presented as mean ±standard
deviation. Body weight, food and water consumption, total
volume of urine, hematology and serum biochemistry, and
absolute and relative organ weights were assumed to be
normally distributed and were analyzed by a one-way analysis
of variance (ANOVA) []. SPSS .K was used for all
statistical analyses (Chicago, IL, USA). A 𝑃 < 0.05 was
considered signicant.
3. Results
3.1. Acute Oral Toxicity Study in Rats. e test substance
was orally administered to a single group of ve male and
vefemaleSDratsatweeksofageindosagelevels
of g/mL/kg B.W. Only one female rat given g/mL/kg
displayed piloerection and activity decrease day aer the
treatment and died within the same day. However, the gross
necropsy results showed no treatment-related changes and
revealed no evidence of specic toxicity related to the test
substance. Also, there was a statistically signicant decrease
in the body weights at day in all test group rats (Tab l e ).
However, the body weights of the test groups recovered.
erefore, this was a temporary side eect from the test
substance. But, all rats except the aforementioned female
rat were not noted in the mortality, clinical, and gross
ndings.
3.2. Acute Oral Toxicity in Beagle Dogs
3.2.1. Clinical Signs and Mortality. No dogs died during
the experimental period. Aer the administration of the
test substance, vomiting was observed. In the male group
given , mg/kg (Table ), very slight vomiting of the test
substanceathrandvomitingofthefoodathrwere
observed in one male. In addition, in the male group given
, mg/kg, severe vomiting was observed in two males at
min, and further vomiting of the food was observed in a
male on day . As shown in Table , vomiting of the food was
observed in one vehicle-treated female on day , moderate
vomiting of the test substance was observed in one female
given , mg/kg at hr, and severe vomiting of the test
substance was observed in two females given , mg/kg
at min. Vomiting of the food was observed in all female
groups on day .
3.2.2. Body Weights. e body weight was observed through-
out the -day experimental period (Table ). In the males, a
loss of body weight was observed in a male given , mg/kg
on days and and in a male given , mg/kg on day .
In the females, a loss of the body weight was observed in a
vehicle-treated female on day , in a female given , mg/kg
ondaysand,andinafemalegiven,mg/kgondays
and .
3.2.3. Necropsy. As shown in Tabl e ,hemorrhageofthe
mucosa of the duodenum and dark red discoloration in
therightlobuleofalungwereobservedinonemale.
However, no abnormalities were observed in other males and
females.
Evidence-Based Complementary and Alternative Medicine
T : Body weight values of Sprague-Dawley rats orally treated with Polygala tenuifolia extracts in acute oral toxicity study.
Group summary of body weight
Dosage in days (Vehicle-treated group, g/ mL/kg/B.W.) (Test substance-treated group, g/ mL/kg/B.W.)
Mean ±S.D. 𝑁Mean ±S.D. 𝑁
Male
−. ±. . ±.
. ±. . ±.
. ±. .∗±.
. ±. .∗±.
. ±. . ±.
Female
−. ±. . ±.
. ±. . ±.
. ±. .∗±.
. ±. . ±.
. ±. . ±.
S.D.: Standard deviation.
𝑁:Numberofanimals.
∗P<. compared to the vehicle-treated group.
(Unit: g).
3.3. Subchronic Oral Toxicity Study in Rats
3.3.1. Clinical Signs and Mortality. Aratwasfounddead
on day and another on day aer administration in
the female group with ,mg/kg/day. In both dead rats,
abnormal fur, dirty noses, and hypothermia were observed
on the day before death. ere were no abnormal ndings
that were associated with the administration of the test
substance other than minor observed signs such as scratched
wounds, crust formation, scarring, and the loss of teeth,
which occurred at a low frequency with no dose-relationship
during the study period (data not shown).
3.3.2. Body Weight and Food and Water Consumption. As a
result of the observations of body weight changes and body
weight gains for weeks during the administration and
observation periods, there were no statistically signicant
changes in all treatment groups of both sexes compared to
the vehicle-treated group. e food consumption in the male
group of mg/kg/day was signicantly increased at weeks
and aer the administration compared to the vehicle-
treated group (Tables and )(𝑃 < 0.05 or 𝑃 < 0.01). ere
was no signicant change in water consumption in all groups
of both sexes compared to the vehicle-treated group.
3.3.3. Ophthalmic Examination. In all groups, there were no
particular signs observed from external eye examinations
at grouping and ophthalmic examinations using a fundus
camera in the nal week of administration (data not shown).
3.3.4. Urinalysis. ere were no particular signs associated
with the administration of the test substance as a result of
urinalysis performed in the nal administration week (data
not shown).
3.3.5. Hematology. e RDW (red cell distribution width)
in the male group administered with , mg/kg/day sig-
nicantly decreased compared to the vehicle-treated group
(Tables and )(𝑃 < 0.01),butnoconsistentchangeswere
observedinthemaleorfemalegroupsinassociationwith
the administration of the test substance in other examination
items. ere were no signicant changes between the groups
as a result of measurement of PT (prothrombin time) and
APTT (activated partial thromboplastin time) (data not
shown).
3.3.6. Serum Biochemistry. As a result of serum biochemical
examinations using the serum of collected blood during
the necropsy (Tables and ), dose-related decreasing
tendencies of ALT, BUN, GLU, AST, ALP, CHO, PRO, and
CPK and dose-related increases of A/G, Na+,andCl
−were
observed in the males. In addition, statistically signicant
dose-dependent decreases of ALT, BUN, and GLU were
observed compared to the vehicle-treated group (𝑃 < 0.05
or 𝑃 < 0.01) and signicant dose-dependent increases of
A/G, Na+,andCl
−were observed compared to the vehicle-
treated group (𝑃 < 0.01).Inthefemalegroups,nostatically
signicant changes were observed in all measurement items.
3.3.7. Necropsy. As a result of the scheduled necropsy of live
animals at weeks aer the administration, no notable dose-
related changes were observed in either males or females. For
males, enlargement of the spleen was observed in one case in
the vehicle-treated group and one case in the me/kg/day-
treated group, a solid brown nodule of the prostate gland
wasobservedinonecaseinthevehicle-treatedgroup,and
diuse red spots or a are-up of the thymus was observed
inonecaseinthevehicle-treatedgroupandonecaseinthe
test substance-treated groups. For the females, the retention
Evidence-Based Complementary and Alternative Medicine
T : Incidence of clinical signs of male beagle dogs orally treated
with Polygala tenuifolia extracts in single dose oral toxicity study.
Sex: male
Day Signs observed Group
, ,
- (. hours) Appe ars normal / ∗/ /
Vomiting (severe) / / /
– ( hour) Appears normal / / /
– ( hours) Appears normal / / /
– ( hours) Appears normal / / /
Vomiting (mild) / / /
– ( hour) Appears normal / / /
– ( hours) Appears normal / / /
Vomiting (food) / / /
– ( hours) Appears normal / / /
Appears normal / / /
Appears normal / / /
Appears normal / / /
Appears normal / / /
Appears normal / / /
Vomiting (food) / / /
Appears normal / / /
Appears normal / / /
Appears normal / / /
Appears normal / / /
Appears normal / / /
Appears normal / / /
Appears normal / / /
Appears normal / / /
Appears normal / / /
∗Number of animals with the sign/number of animals examined.
of a clear uid in the uterus was observed in the vehicle-
treatedgroupandthe,,and,mg/kg/day-treated
groups(,,,andcases,resp.)anddiuseredspotsinthe
thymus and dark yellowish brown discoloration of the lung
were observed in the mg/kg/day-treated group in two
cases and one case, respectively. Additionally, enlargement of
thesubmandibularlymphnodewasobservedinonecasein
the,mg/kg/day-treatedgroup.Asaresultofthenecropsy
of two dead females of the , mg/kg/day-treated group,
diuse red spots or dark red coloration in the lung and diuse
red spots in the anterior stomach and in the thymus were
observed along with gas retention due to postmortal changes
(data not shown).
3.3.8. Organ Weight. For males, the absolute and relative
weights of the thymus showed signicant increases in the
and mg/kg/day-treated groups compared to the vehicle-
treated group (Tables and )(𝑃 < 0.05 or 𝑃 < 0.01)and
therelativeweightoftheleepididymisshowedasignicant
decrease in the mg/kg/day-treated group compared to the
vehicle-treated group (𝑃 < 0.05), but these changes were
not dose-related. For females, the relative weight of the liver
T : Incidence of clinical signs of female beagle dogs orally
treated with Polygala tenuifolia extracts in single dose oral toxicity
study.
Sex: female
Day Signs observed Group
, ,
- (. hours) App ears normal /∗/ /
Vomiting (severe) / / /
– ( hour) Appears normal / / /
Vomiting (moderate) / / /
– ( hours) Appears normal / / /
– ( hours) Appears normal / / /
– ( hours) Appears normal / / /
– ( hours) Appears normal / / /
– ( hours) Appears normal / / /
Appears normal / / /
Vomiting (food) / / /
Appears normal / / /
Appears normal / / /
Appears normal / / /
Appears normal / / /
Appears normal / / /
Vomiting (food) / / /
Appears normal / / /
Appears normal / / /
Appears normal / / /
Appears normal / / /
Appears normal / / /
Appears normal / / /
Appears normal / / /
∗Number of animals with the sign/number of animals examined.
showed a signicant increase in the , mg/kg/day-treated
group compared to the vehicle-treated group (𝑃 < 0.05).
3.3.9. Histopathology. As a result of histopathological exami-
nation of rats with a scheduled necropsy, in the males in the
vehicle-treated group, severe granulomatous inammation of
the prostate gland, moderate pyelonephritis in the kidney,
sightcystsinthepituitarygland,andmoderategranuloma
in the liver were observed in one case, along with moderate
inammatory cell inltration in the Hadrian gland and slight
inammatory cell inltration in the heart (one case each).
For the females, two cases of slight tubular regeneration in
thekidneywereobservedinonecaseeach.Inthemales
with a scheduled necropsy from the , mg/kg/day-treated
group, slight tubular remnants in the pituitary gland, slight
granuloma in the liver, and a slight dilation of the lumen in
the uterus were observed in one case in each group. In the
low-andmedium-dosegroups,paleyellowishbrowndiscol-
orationofthelelobeofthelungs(identiedasfocalalve-
olitis) was observed in one female from the mg/kg/day-
treated group, and diuse red spots in the thymus were
observed in two cases (identied as focal hemorrhages). No
abnormal ndings were observed in the abnormal organs.
Evidence-Based Complementary and Alternative Medicine
T:BodyweightvaluesofbeagledogsorallytreatedwithPolygala tenuifolia extracts in single dose oral toxicity study.
Sex: male
Group Body weights (kg)
(mg/kg) Animal ID d d d d d Gain
(𝑛=2)
. . . . . .
. . . . . .
Mean . ±. . ±. . ±. . ±. . ±. . ±.
,
(𝑛=2)
. . . . . .
. . . . . .
Mean . ±. . ±. . ±. . ±. . ±. . ±.
,
(𝑛=2)
. . . . . .
. . . . . .
Mean . ±. . ±. . ±. . ±. . ±. . ±.
Sex: female
Group Body weights (kg)
(mg/kg) Animal ID d d d d d Gain
(𝑛=2)
. . . . . .
. . . . . .
Mean . ±. . ±. . ±. . ±. . ±. . ±.
,
(𝑛=2)
. . . . . −.
. . . . . −.
Mean . ±. . ±. . ±. . ±. . ±. −. ±.
,
(𝑛=2)
. . . . . .
. . . . . .
Mean . ±. . ±. . ±. . ±. . ±. . ±.
T : Gross ndings of beagle dogs orally treated with Polygala tenuifolia extracts in single dose oral toxicity study.
Sex: male
Group Gross observation Frequency
(mg/kg) Location Gross ndings Death Survivors
No gross ndings ∗/ /
, No gross ndings / /
No gross ndings / /
, Duodenum Hemorrhage on mucous membrane / /
Lung Dark red discoloration of right lobule / /
Sex: female
Group Gross observation Frequency
(mg/kg) Location Gross ndings Death Survivors
No gross ndings ∗/ /
, No gross ndings / /
, No gross ndings / /
∗Number of animals with the sign/Number of animals examined.
As a result of the histopathological examinations on the two
dead female rats from the , mg/kg/day-treated group,
it was found that the rats had suered severe postmortal
changes, and therefore an accurate evaluation of the lesions
wasdicult.However,inratsthatdiedondayaer
the administration, diuse vacuolation in the adrenal gland,
atrophy of the spleen, and inammatory cell inltration in
the heart were observed to a minor extent, and no abnormal
ndings were observed in a dead animal on day aer the
administration.
4. Discussion
Polygala tenuifolia root is a famous traditional medicine.
Althoughmanystudieshavereportedthepharmacological
ecacy of the root extract, there is no information on its
safety, such as its acute and subchronic oral toxicity. First, we
investigated the acute and subchronic oral toxicity of the root
extract in rats. e root extract was administered orally at
or g/kg body weight for the acute oral toxicity test and at
, , , or , mg/kg body weight for the subchronic
oral toxicity test. In the acute oral toxicity study, one female
Evidence-Based Complementary and Alternative Medicine
T : Food consumption of male rats.
Food consumption (g)
Study: -RR- Dose (mg/kg/day) Sex: male
Weeks ,
Mean ±S.D. 𝑁Mean ±S.D. 𝑁Mean ±S.D. 𝑁Mean ±S.D. 𝑁
.±. . ±. . ±. . ±.
. ±. . ±. . ±. . ±.
. ±. . ±. . ±. . ±.
. ±. . ±. . ±. . ±.
. ±. . ±.∗∗ . ±. . ±.
.±. . ±. . ±. . ±.
. ±. . ±. . ±. . ±.
.±. . ±. . ±. . ±.
.±. . ±. . ±. . ±.
.±. . ±. . ±. . ±.
. ±. . ±.∗ . ±. . ±.
. ±. . ±. . ±. . ±.
. ±. . ±. . ±. . ±.
. ±. . ±. . ±. . ±.
∗P<. and ∗∗P<. compared to the vehicle-treated group.
T : Food consumption of female rats.
Food consumption (g)
Study: -RR- Dose (mg/kg/day) Sex: female
Weeks ,
Mean ±S.D. 𝑁Mean ±S.D. 𝑁Mean ±S.D. 𝑁Mean ±S.D. 𝑁
. ±. . ±. . ±. . ±.
.±. . ±. . ±. . ±.
.±. . ±. . ±. . ±.
.±. . ±. . ±. . ±.
.±. . ±. . ±. . ±.
.±. . ±. . ±. . ±.
.±. . ±. . ±. . ±.
.±. . ±. . ±. . ±.
. ±. . ±. . ±. . ±.
.±. . ±. . ±. . ±.
. ±. . ±. . ±. . ±.
. ±. . ±. . ±. . ±.
. ±. . ±. . ±. . ±.
. ±. . ±. . ±. . ±.
rat given g/kg of the root extract exhibited piloerection
and decrease in activity levels day aer treatment and died
withinthesameday.egrossndingsoftheautopsyshowed
no evidence of specic toxicity related to the test substance. In
addition, there was a statistically signicant decrease in body
weight at day in all test-group rats, but the body weights
later recovered. Hence, this nding may show a temporary
eect of the test substance. However, all rats except the afore-
mentionedfemaleratwerenotnotedinthemortality,clinical,
and gross ndings. ese results suggest that the approximate
lethal dose for the root extract of P. t e n u i f o l i a may be ≥g/kg
in male rats and ≤ g/kg in female rats under the conditions
used in this study. In the subchronic oral toxicity study,
there were no abnormal signs associated with administration
of the test substance. No specic ndings for body weight,
consumption of food and water, ophthalmic examination,
urinalyses, and blood clotting time were observed in relation
to administration of the test substance. In the female group
of , mg/kg/day, rats that died on day and day
showed normal clinical signs, body weight changes, and
consumption of food and water during the study period
but demonstrated abnormal fur, dirty noses or mouths, and
hypothermia the day before death. Autopsies revealed dark
redcolorationordiuseredspotsinthelungs,aswell
Evidence-Based Complementary and Alternative Medicine
T : Hematological values (mean ±standard deviation) of male rats in the subchronic oral toxicity study of the root of Polygala tenuifolia.
Parameter Dose (mg/kg/day)
,
WBC (/𝜇L) . ±. . ±. . ±. . ±.
RBC (/𝜇L) . ±. . ±. . ±. . ±.
Hemoglobin (g/dL) . ±. . ±. . ±. . ±.
Hematocrit (%) . ±. . ±. . ±. . ±.
MCV (fL) . ±. . ±. . ±. . ±.
MCH (pg) . ±. . ±. . ±. . ±.
MCHC (g/𝜇L) . ±. . ±. . ±. . ±.
RDW (%) . ±. . ±. . ±. . ±.∗∗
HDW (g/dL) . ±. . ±. . ±. . ±.
Platelet (/𝜇L) . ±. . ±. . ±. . ±.
MPV (fL) . ±. . ±. . ±. . ±.
Reticulocyte (%) . ±. . ±. . ±. . ±.
𝑁
WBC: white blood cell count; RBC: red blood cell count; MCV: mean corpuscular volume; MCH: mean corpuscular hemoglobin concentration; MCHC; mean
corpuscular hemoglobin concentration; RDW: red cell distribution width; HDW: hemoglobin distribution width; MPV: mean platelet volume.
∗∗Signicant dierence at P<. compared with the vehicle-treated group.
T : Hematologicalvalues (mean ±standard deviation) of female rats in the subchronic oral toxicity study of the root of Polygala tenuifolia.
Parameter Dose (mg/kg/day)
,
WBC (/𝜇L) . ±. . ±. . ±. . ±.
RBC (/𝜇L) . ±. . ±. . ±. . ±.
Hemoglobin (g/dL) . ±. . ±. . ±. . ±.
Hematocrit (%) . ±. . ±. . ±. . ±.
MCV (fL) . ±. . ±. . ±. . ±.
MCH (pg) . ±. . ±. . ±. . ±.
MCHC (g/𝜇L) . ±. . ±. . ±. . ±.
RDW (%) . ±. . ±. . ±. . ±.
HDW (g/dL) . ±. . ±. . ±. . ±.
Platelet (/𝜇L) . ±. . ±. . ±. . ±.
MPV (fL) . ±. . ±. . ±. . ±.
Reticulocyte (%) . ±. . ±. . ±. . ±.
𝑁
WBC: white blood cell count; RBC: red blood cell count; MCV: mean corpuscular volume; MCH: mean corpuscular hemoglobin concentration; MCHC: mean
corpuscular hemoglobin concentration; RDW: red cell distribution width; HDW: hemoglobin distribution width; MPV: mean platelet volume.
as diuse red spots in the anterior stomach and thymus
gland. A more accurate evaluation of the lesions was made
dicult by severe postmortem changes in two rats during
the histopathological studies. In the rat that died days
aer administration of the root extract, diuse vacuolation
in the adrenal glands, atrophy of the spleen, and inltration
of the inammatory cells in the heart were observed. In
the rat that died on day , abnormal ndings were not
observed.Asmentionedabove,theclinicalsigns,bodyweight
changes, food and water consumption, necropsy ndings,
and histopathological ndings obtained for the dead rats did
not indicate any toxicity of the test substance. erefore, the
deaths of the rats were not associated with the administration
of the test substance and were judged to be accidental due
to administration errors. is hypothesis is supported by the
fact that no abnormal ndings were observed in relation to
the administration of the test substance in the remaining
rats in the same dose group. We demonstrated that the
test substance could be used as a food supplement, because
some famous food ingredients were toxic at high doses, (e.g.,
lemongrass (Cymbopogon citratus)essentialoil,Roystonea
regia fruit, and 𝛼-glycerylphosphorylcholine (AGPC)) [–
]. e results of the serum biochemical examinations of the
rat sera revealed reductions in the levels of AST and ALT in
themalegroups.ASTandALTarecoenzymesofpyridoxal
phosphate (PALP). PALP is a derivative of vitamin B6and
ASTandALTareactivatedbyunionwithPALP,butnon-
linked aminotransferases are not activated. Hence, reductions
inthelevelsofALTandASToccurifthereislowgeneration
of aminotransferases or PALP or if the linkage between the
Evidence-Based Complementary and Alternative Medicine
T : Serum biochemical values (mean ±standard deviation) of male rats in the subchronic oral dose toxicity study of the root of Polygala
tenuifolia.
Parameter Dose (mg/kg/day)
,
Number of animals examined
AST (IU/L) . ±. . ±. . ±. . ±.
ALT (IU/L) . ±. . ±. . ±.∗. ±.∗∗
ALP (IU/L) . ±. . ±. . ±. . ±.
BUN (mg/dL) . ±. . ±.∗. ±.∗. ±.∗
CRE (mg/dL) . ±. . ±. . ±. . ±.
GLU (mg/dL) . ±. . ±.∗∗ . ±.∗. ±.∗
CHO (mg/dL) . ±. . ±. . ±. . ±.
PRO (g/dL) . ±. . ±. . ±. . ±.
CPK (IU/L) . ±. . ±. . ±. . ±.
ALB (g/dL) . ±. . ±. . ±. . ±.
BIL (mg/dL) . ±. . ±. . ±. . ±.
TG (mg/dL) . ±. . ±. . ±. . ±.
IP (mg/dL) . ±. . ±. . ±. . ±.
Ca+ (mg/dL) . ±. . ±. . ±. . ±.
A/G (ratio) . ±. . ±.∗∗ . ±.∗∗ . ±.∗∗
Na+(mmol/L) . ±. . ±.∗∗ . ±.∗∗ . ±.∗∗
K+(mmol/L) . ±. . ±. . ±. . ±.
Cl−(mmol/L) . ±. . ±.∗∗ . ±.∗∗ . ±.∗∗
𝑁
∗/∗∗Signicant dierence at P<./. compared with the vehicle-treated group.
T : Serum biochemical values (mean ±standard deviation) of female rats in the subchronic oral dose toxicity study of the root of
Polygala tenuifolia.
Parameter Dose (mg/kg/day)
,
Number of animals examined
AST (IU/L) . ±. . ±. . ±. . ±.
ALT (IU/L) . ±. . ±. . ±. . ±.
ALP (IU/L) . ±. . ±. . ±. . ±.
BUN (mg/dL) . ±. . ±. . ±. . ±.
CRE (mg/dL) . ±. . ±. . ±. . ±.
GLU (mg/dL) . ±. . ±. . ±. . ±.
CHO (mg/dL) . ±. . ±. . ±. . ±.
PRO (g/dL) . ±. . ±. . ±. . ±.
CPK (IU/L) . ±. . ±. . ±. . ±.
ALB (g/dL) . ±. . ±. . ±. . ±.
BIL (mg/dL) . ±. . ±. . ±. . ±.
TG (mg/dL) . ±. . ±. . ±. . ±.
IP (mg/dL) . ±. . ±. . ±. . ±.
Ca+ (mg/dL) . ±. . ±. . ±. . ±.
A/G (ratio) . ±. . ±. . ±. . ±.
Na+(mmol/L) . ±. . ±. . ±. . ±.
K+(mmol/L) . ±. . ±. . ±. . ±.
Cl−(mmol/L) . ±. . ±. . ±. . ±.
𝑁
Evidence-Based Complementary and Alternative Medicine
T : Absolute and relative organ weight of males in the subchronic oral dose toxicity study of the root of Polygala tenuifolia.
Parameter Dose (mg/kg/day)
,
Body weight . ±. . ±. . ±. . ±.
Adrenal gland, le . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Adrenal gland, right . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Pituitary gland . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
ymus . ±. . ±.∗∗ . ±.∗∗ . ±.
Per body weight (%) . ±. . ±.∗. ±.∗∗ . ±.
Prostate . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Testis, le . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Testis, le . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Epididymis, le . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Epididymis, right . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Spleen . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Kidney, le . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Kidney, right . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Heart , ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Lung . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Brain . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Liver . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
𝑁
Body weight before necropsy and aer fasting.
∗/∗∗Signicant dierence at P<./. compared with the vehicle-treated group.
aminotransferases and PALP is disturbed. Cephalosporin-
like antibiotics are known to induce reductions in ALT levels,
but in toxicity studies, reductions have been mentioned in
general and toxicological signicance has not been attributed
[,]. e blood urea nitrogen (BUN) levels in the blood
represent the nitrogen content in relation to urea, which
is a metabolite of nitrogen and is closely related to heart
diseases. e BUN value might show a decrease, due to a
reduction in urea synthesis during toxic hepatitis, severe hep-
atic impairment, and consuming a low protein diet. However,
in the present study, no abnormal ndings related to dose
were observed. In general, the decrease in BUN level is not a
critical factor in toxicity studies []. Moreover, these changes
were not regarded as adverse eects because they remained
within normal ranges [,]. Hematological examinations
revealed reductions in red blood cell volume distribution
width(RDW)inthemalegroupgiven,mg/kg/day.
Decreasing tendencies in the levels of GLU, ALP, CHO, PRO,
ALB, and CPK and increments in the levels of A/G, Na+,
and Cl−in male treatment groups were within the normal
range [,] and also remained within the normal range
when compared with the historical data []availableat
ChemOn Inc. In the vehicle-treated group, increments or
reductions in some measurement items were also observed
accidentally and were thought to be somewhat contributory
to the above results []; therefore, it was considered that
these changes were toxicological and were not associated with
the administration of the test substance. RDW decreased
Evidence-Based Complementary and Alternative Medicine
T : Absolute and relative organ weight of females in the subchronic oral dose toxicity study of the root of Polygala tenuifolia.
Parameter Dose (mg/kg/day)
,
Body weight . ±. . ±. . ±. . ±.
Ovary, le (g) . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Ovary, right . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Adrenal gland, le . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Adrenal gland, right . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Pituitary gland . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
ymus . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Uterus . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Spleen . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Kidney, le . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Kidney, right . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Heart . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Lung . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Brain . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.
Liver . ±. . ±. . ±. . ±.
Per body weight (%) . ±. . ±. . ±. . ±.∗
𝑁
Body weight before necropsy and aer fasting.
∗Signicant dierence at P<. compared with the vehicle-treated group.
signicantly, but the changes remained within the limits
of normal biological variations [,]. Signicant changes
intheabsolute/relativeweightsofthethymusglandin
themalegroupsgivenandmg/kg/day,therelative
weightsoftheleepididymisinthemalegroupsgiven
mg/kg/day, and the relative weights of the liver in the
female group given , mg/kg/day were not accompanied
by abnormal autopsy or histopathological ndings. A rela-
tionshipwithsexordosewasnotobserved,sothesechanges
were not considered to be a toxicological feature of the test
substance. Even though the weights of the thymus gland
and le epididymis changed signicantly, they were within
the limits of normal biological variations; this nding was
in accordance with results reported by Wang et al. [].
Regardingrelativeorganweight,anincreaseintheliverinthe
female rats, compared with Banpungtongseong-san reported
organweightsanalysis,showedthattherewerestatistically
signicant changes in the absolute weights of the thymus in
themalegroupof,mg/kg/day.However,thesechanges
were not regarded as adverse eects because they were minor,
remained within normal historical control ranges for Fischer
rats, and were not correlated with pathological lesions in
therespectiveorgans[,]. Autopsy and histopathological
examinations revealed abnormal ndings in some organs, but
the prevalence of these ndings was low and these changes
were spontaneous and nonspecic [,]. erefore, the
toxicological importance of these ndings was negligible.
No abnormal ndings were observed in association with
administration of the test substance upon histopathological
examination of abnormal organs in low- and medium-dose
groups.
Second, acute oral toxicity of the root extract P. t e n u i f o -
lia in beagle dogs was investigated. e root extract was
administered orally at , or ,mg/kg body weights;
the vehicle-treated group was only given gelatin capsules for
the single dose oral toxicity test. As a result of the present
Evidence-Based Complementary and Alternative Medicine
study, vomiting and inhibition of body weight gain were
observed aer the administration of the test substance, but
other abnormalities in clinical signs, deaths, and abnormal
ndings in the necropsy results were not observed in relation
to administration of the test substance. e results of clinical
sign examinations of the dogs observed vomiting in one male
given,mg/kgathrandinonemalegiven,mg/kg
on day . is was aer the administration was considered
to be unrelated to the test substance because vomiting was
also observed in the vehicle-treated female group on day
, and vomiting like this is commonly observed in dogs
which have a well-developed vomiting system. However,
these changes were not considered serious when compared
with the previous study, as some famous oriental medicine
and food compositions were found to be toxic, including
Paecilomyces sinclairii [], SH-B [], epigallocatechin
gallate (EGCG) [], novel thiazolidinedione (MCC-)
[], and nelumbinis semen (NS, the seeds of Nelumbo
nucifera)[]. Vomiting of the food occurred sporadically
in some dogs in all groups, including the controls. is
frequency was typical for this colony of dogs []. In general,
the incidence of vomiting did not suggest an eect of the
test substance []. Vomiting is a common presenting sign
in small animal practice []. Moreover, many peripheral
stimuli of abdominal structures will initiate vomiting in dogs
[,]. e “chemoreceptor trigger zone” of the brainstem
hasbeenidentiedastheareapostremathatislocatedon
the dorsal surface of the medulla oblongata adjacent to the
caudal end of the fourth ventricle []. erefore, it was
not considered to be related to the treatment with the test
substance. e loss of body weight was observed in males and
females, but because vomiting of the test substance occurred
more frequently at ,mg/kg than at , mg/kg, the
absorption of test substance seemed to be relatively higher
at , mg/kg than at , mg/kg. Also, the loss of body
weight showed a temporary serious tendency on days and
aer the administration. However, the body weights of
the male , mg/kg and , mg/kg groups recovered.
In addition, the vehicle-treated group of females showed a
tendency to decrease in body weight from approximately
days. In all groups, increment and reduction tendencies
were also observed, and these changes were observed [–
].echangesinbodyweightswereprobablytheresult
of irritation of the alimentary tract []. We also considered
gastroenteric issues from vomiting as accordingly inducing
a disorder of the nutriments absorption []. A hemorrhage
of the mucosa of the duodenum and the dark discoloration
of the lung observed in a male given , mg/kg were not
considered as test substance-related changes, because these
were occasionally found in domestic beagle dogs without
administration of any of the test substances. Dog experiment
ndings were generally observed as environmental causes,
because these results are ignored []. In the necropsy results,
adarkredspotonthelungandalightgraycolorchange
were observed. However, this change was not dose-related
[],butitwasanaccidentalchange.Also,thelungsshowed
discoloration(dark,mottled,andfocus),andthestomach
showed discoloration [,]. e gross lesion of the mass
or nodule corresponded to the microscopic lesion of the
thrombus. ese were direct eects of the drug or were
secondary to the vascular damage caused by dehydration
(with hemodynamic shock) from vomiting and diarrhea [].
Histopathology changes and hemorrhages in the lung were
observed by Werley et al. []. However, they were considered
to be typical of spontaneously arising background ndings,
and no other biologically signicant eects were observed
by histopathology studies on the tissues and organs. e
discolorations in the other various organs and tissues were
within the range of normal background lesions in dogs of this
strain and age and were therefore not considered to be related
to the test substance [].
In this experiment, the administration of the root extract
of Polygala tenuifolia Willdenow ( g/kg/day) did not result in
acute oral toxicity in the SD rats. e minimum lethal dose
(MLD) was also considered to be higher than , mg/kg,
since vomiting of the test substance occurred at , mg/kg.
In the subchronic oral toxicity test, no obvious toxic changes
due to administration of the root extract of P. t e n u i f o l i a
were observed in any of the parameters tested (clinical
signs, mortality, body weight changes, food and water con-
sumption, ophthalmic examination, urinalyses, hematology,
serum biochemistry, organ weights, autopsy, and histopathol-
ogy). erefore, under the experimental conditions, the no-
observed-adverse-eect level (NOAEL) of the root extract of
P. t e n u i f o l i a was determined to be , mg/kg/day for both
sexes, but the target organ was not established. In conclusion,
thepresentstudydemonstratedthattheacuteorsubchronic
toxicity of the root extract of Polygala tenuifolia was not toxic
in rats and beagle dogs.
Conflict of Interests
e authors of this paper have no conict of interests.
Acknowledgments
e collection, analysis, and interpretation of data of the
acute toxicity and the subchronic toxicity were supported
by SNUH-GLP (Non-Clinical Research Center, Biomedical
Research Institute, Seoul National University Hospital) and
Preclinical Research Center, ChemOn Inc., respectively.
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