521

Journal of Health Science, 57(6) 521–531 (2011)— Regular Article —Preliminary Research for the Anti-obesity Effect of Puerariae
Flos Extract in Humans
Tomoyasu Kamiya,∗, a Yuki Matsuzuka, a Nobutaka Kusaba, a Motoya Ikeguchi, a
Kinya Takagaki, a and Kazuo Kondo baResearch and Development Division, Toyo Shinyaku Co. Ltd., 7–28 Yayoigaoka, Tosu-shi, Saga 841–0005, Japan and b Institution ofEnvironmental Science for Human Life, Ochanomizu University, 2–1–1 Ohtsuka, Bunkyo-ku, Tokyo 112–8610, Japan(Received June 10, 2011; Accepted August 12, 2011; Published online August 19, 2011)

This research is preliminary study to examine whether puerariae flos extract (PFE) makes any action to body fat
of man or not. We conducted the double-blind placebo controlled study on eighty mildly obese subjects for 8 weeks.
The subjects were randomly divided into 4 groups: I, II, III, and IV. Subjects in groups I, II, and III consumed test
food containing 100, 200, and 300 mg PFE, respectively, while those in group IV were given placebo food for
8 weeks. All subjects were instructed to restrict their total-energy intake to within 2650 kcal/day in the case ofmales and 2300 kcal/day in the case of females during the test period. Haematological and biochemical markersof blood, urinary markers, and physical markers were examined at 0, 4, and 8 weeks during the test period. As
a result, no adverse effects were noted in any of the groups. For physical examinations, we analysed only males
whose initial body mass index (BMI) values were over 24. BMI value and body weights of the subjects in group
III were significantly reduced, and total fat area and subcutaneous fat area of the subjects in groups II and III weresignificantly reduced over the 8-week test period. Moreover, as a result of comparison among groups by unpaired
t-test, significant difference appeared between group III and group IV in BMI. Total fat area in group III tend to
decrease compared with group IV. By these results, the reduction of BMI in group III may reflect fat area reduction.
To confirm the anti-obesity effect of PFE, we need to research particularly by extensive studies hereafter.
Key words —— anti-obesity, puerariae flower extract, kudzu, isoflavone, abdominal fat area

INTRODUCTION
Obesity is a major public health issue, as it isclosely related to mortality. Obesity is caused bya variety of factors, one of which is an imbalance
between energy intake and expenditure. Therefore,
it is important for the prevention of obesity to suppress energy intake and increase its expenditure.
Diet therapy is generally advocated for suppressing
energy intake, while exercise is recommended for
ensuring adequate energy expenditure. In addition,
the use of a dietary supplement that can reduce energy intake by inhibiting the absorption of sugar andfat and/or elevate energy metabolism in the liver andmuscles via β-oxidation is recommended.
∗

To whom correspondence should be addressed: Research andDevelopment Division, Toyo Shinyaku Co. Ltd., 7–28 Yayoigaoka, Tosu-shi, Saga 841–0005, Japan. Tel.: +81-942-813555; Fax: +81-942-81-3554; E-mail: kamiyat@toyoshinyaku.
co.jp

Pueraria flos extract (PFE) is an extract fromflowers of kudzu (Puerariae thomsonii). Kudzu, aleguminous plant distributed in Japan, China, andother areas, has long been used in folk medicine.
In particular, the Puerariae flower is used inJapanese and Chinese folk medicine for curinghangovers.1–3)
Niiho et al. confirmed that the consumption of
the Puerariae lobata flower reduces glutamic oxaloacetic transaminase (GOT) or glutamic pyruvic
transaminase (GPT) levels in individuals with liver
injury due to carbon tetrachloride (CCl4 ) or a highfat diet in animal study.1, 2) In East Asia, this flower
has also recently been used in herbal medicines forthe management of menopausal symptoms.4, 5)
Puerariae thomsonii is known to contain
7 isoflavones (4 isoflavone glucosides: tectoridin, tectorigenin 7-O-xylosylglucoside, 6hydroxygenistein-6,7-diglucoside, and glycitin; and3 aglycones: tectorigenin, glycitein, and genistein)C 2011 The Pharmaceutical Society of Japan

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Table 1. Analysis Value of Nutrient Composition for Test Food (1.0 g per Packet)

Energya) (kcal)
Moisture (g)Proteinb) (g)Fat (g)Carbohydrate (g)Test food(containing 100 mgPFE)
3.94
0.036
0.019
0.033
0.891

Test food(containing 200 mgPFE)
3.88
0.039
0.039
0.036
0.851

Test food(containing 300 mgPFE)
3.84
0.040
0.059
0.040
0.812

Placebo food
3.96
0.037
0.001
0.031
0.918

a) Calorie conversion factor: Protein 4, Fat 9, Carbohydrate 4. b) Nitrogen-to-protein conversion factor: 6.25.

and 3 saponins (soyasaponin I, kaikasaponin III,and kakkasaponin I).6)
It has been reported that isoflavones such as
genistein affect lipid metabolism in the liver and inhibit adipocyte differentiation,7, 8) and that saponins
have an inhibitory effect on lipase activity.9) Therefore, PFE consumption is expected to provide thedual benefits of reducing energy intake and increasing its expenditure.
Wang et al. confirmed that flavones derived
from puerariae radix have inhibitory effects on bodyweight, abdominal fat, and lipids in the liver.10)
However, the impact of PFE on body fat is notknown. This clinical study was preliminary conducted on eighty mildly obese subjects to view the
body fat reducing action of PFE.

MATERIALS AND METHODS
Design and Subjects —— We had drew up a clinical protocol for this study. Then this study had been
approved by Institutional Review Board of Kurume
clinical pharmacology clinic (July 28, 2005) before
its launch. Furthermore this study was conducted inaccordance with the Helsinki Declaration, and informed consent was obtained from all subjects. We
entrusted all affairs of this study to Iberica Holdings Co., Ltd. (Fukuoka, Japan). They carried it out
at Kurume clinical pharmacology clinic (Kurume,
Japan).
Volunteers who live in around Kurume city had
a questionnaire related to previous disease and administration of drugs, blood tests, body checks anda medical interview by a doctor. Subjects aged between 20 and 65 years were recruited for the study,
according to fixed inclusion and exclusion criteria. The inclusion criteria pertained to the following prospective subjects: those having a body massindex (BMI) value of 23–30 or a waist circumference larger than 85 cm for males or 90 cm for fe-

males, those who had visited a hospital for treatment
and provided informed consent for study enrolment,
those who were deemed to be suitable for the study
by a doctor, those who had been certified as healthy
by a doctor during a prior clinical inspection, andthose who were not under any drug treatment.
The exclusion criteria pertained to the following
prospective subjects: those using functional foods,
cosmetics, or instruments that may influence lipid
metabolism (e.g., hypolipidaemic compounds andantihypertensive agents), those with liver dysfunction, pregnant women or those suspected to be pregnant, those who were considered unsuitable for the
study due to an illness or the possibility of developing serious side effects, and those who were deemed
to be inappropriate subjects by a doctor.
Test Food —— Tablets containing PFE (Ohta’s Isan
Co. Ltd., Ushiku, Japan), reduced palatinose, cellulose, fatty acid esters of sucrose, and silicon dioxide were used as the test food. The placebo foodconsisted of tablets containing caramel as a dye, instead of PFE. Table 1 shows analysis value of nutrient composition. The test food and the placebo
food were distributed with plain aluminium packets
sealed individually containing 1.0 g per packet. The
subjects in each group were required to consume theallotted tablet once per day.
Experimental Design —— Double-blind placebo
controlled study was conducted in this case. Figure 1 shows the test schedule. Following a 2-week
pre-observation period (i.e., at “week 0”), haematological, urine, and physical examinations and bloodbiochemical analyses were conducted at the hospital. Further, 80 subjects with mild obesity were randomly divided into 4 groups using block randomization. Subjects in groups I, II, and III were required
to ingest 100, 200, and 300 mg of PFE, respectively,
while those in group IV were required to ingest
the placebo food once per day for 8 weeks. Subjects were restricted to take medicines and supplements as a general rule and had dietary instruction

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No. 6

Fig. 1. Test Schedule

to prevent from rapid change of calories consumed
during the intake of PFE. We distributed pedometers to the subjects to instruct not to make rapidchange to their amount of exercise. Regarding otherdaily habit, we instructed to have a life as usual.
Subjects were also asked to keep the daily recordof test food consumption, rational symptoms, allfoods consumed including articles of taste, amountof exercise and intake of prohibited medicinesor supplements.
Examinations were conducted at 4 and 8 weeks
after week 0 (i.e., during week 4 and week 8).
Blood biochemical analyses and physical examinations were conducted at week 4 and additional
haematological and urine examinations and bloodbiochemical analyses were conducted at week 8.
Furthermore, subjective symptoms wererecorded 2 weeks after the study period.
Dietary Instruction —— The subjects were instructed to limit their total energy intake to under 2650 kcal/day in the case of males, and 2300
kcal/day in the case of females, and also instructed
not to reduce food intake substantially basing on the
Dietary Reference Intakes for Japanese, 2005. The
details of their meal intake are provided as followings. Breakfast: bread or rice, a side dish, and milk,
Lunch: nutritious foods for which the total energy
intake could be measured and the nutrients classified, and Dinner: a home-delivered meal (main dish,
300 kcal; side dish, 320 kcal).
With regard to alcohol consumption, the subjects were permitted less than 1 large bottle of beeror the equivalent. Alcohol consumption was forbidden on the day before a blood examination or med-

ical interview.
Haematological Examinations and Biochemical
Examinations of Blood —— Haematological examinations were conducted at weeks 0 and 8. Bloodbiochemical analyses were conducted at weeks 0, 4,
and 8.
The parameters assessed in the haematological examinations were the white blood cell (WBC)count, red blood cell (RBC) count, haemoglobin(Hb) level, and haematocrit (Ht) value. Those assessed in the blood biochemical analyses performed
at week 4 were the triglyceride, total cholesterol(T-Cho), low-density-lipoprotein cholesterol (LDLCho), high-density-lipoprotein cholesterol (HDLCho), remnant-lipoprotein cholesterol (RLP-Cho),leptin, fasting blood glucose (FBG), and insulin levels. In addition, the parameters assessed in the
blood biochemical analyses performed at week 8
were the total proteins, and the GOT, GPT, γglutamyl transpeptidase (γ-GTP), albumin, creatinine, total bilirubin (TB), blood urea nitrogen(BUN), uric acid, creatine kinase (CPK), alkaline
phosphatase (ALP), lactate dehydrogenase (LDH),
triglyceride, T-Cho, LDL-Cho, HDL-Cho, RLPCho, leptin, FBG, insulin, and haemoglobin A1c(HbA1c) levels.
Urine Examinations —— Urine examinations
were conducted at weeks 0 and 8. The parametersassessed were the urine sugar and protein content
and the presence of blood in the urine.
Rational Symptoms —— Subjective symptoms,
which occurred during the study, were tallied.
Symptoms, which confirmed a relation to the test
food by a doctor, were also tallied.

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Vol. 57 (2011)

Table 2. Subject Characteristics

NumberAge (year-old)
Body weight (kg)BMI (kg/m2 )Waist circumference (cm)Hip circumference (cm)
Total fat area (cm2 )Visceral fat area (cm2 )Subcutaneous fat area (cm2 )Systolic blood pressure (mmHg)
Diastolic blood pressure (mmHg)

Group I(n = 18)
Male: 16
Female: 2
40.3 ± 7.975.2 ± 9.1
26.7 ± 2.6
95.2 ± 8.5
99.9 ± 5.4
343.0 ± 82.4
128.5 ± 38.1
214.5 ± 65.1
128.9 ± 11.4
82.2 ± 7.3Group II(n = 15)
Male: 12
Female: 337.9 ± 9.1
74.2 ± 11.8
26.4 ± 3.2
95.2 ± 10.1
98.6 ± 5.7
323.0 ± 126.4
133.8 ± 115.6
189.2 ± 52.4
128.5 ± 17.3
80.3 ± 15.2

Group III(n = 18)
Male: 15
Female: 3
39.3 ± 12.4
72.0 ± 9.4
25.5 ± 2.9
93.3 ± 7.6
99.8 ± 4.4
306.6 ± 92.0
111.3 ± 44.2
195.3 ± 65.2
120.1 ± 9.4
74.9 ± 9.6Group IV(n = 18)
Male: 15
Female: 336.7 ± 9.4
76.1 ± 8.6
26.5 ± 2.5
96.8 ± 8.4
101.4 ± 4.2
331.4 ± 85.4
122.8 ± 53.5
208.6 ± 59.4
126.1 ± 15.4
80.8 ± 12.6

See Fig. 1 for Groups. Values are means ± S.D.

Physical Examinations —— Physical examinations were conducted at weeks 0 and 8, and theparameters assessed were height (data not shown),body weight, BMI, waist circumference, hip circumference, total fat area, visceral fat area, subcutaneous fat area, systolic blood pressure, and diastolicblood pressure.
The hip circumference was measured using acalibrated tape. The waist circumference, total fat
area, visceral fat area, and subcutaneous fat area
were measured by performing computerized tomography (CT) while the subjects held their breath following expiration. The CT scan data were analysedusing the Fat scan software (East Japan Institute ofTechnology Co. Ltd., Hitachi, Japan).11)
Statistical Analysis —— We performed a repeatedmeasures one-way analysis of variance (ANOVA) toinvestigate the changes over the time course. Whenstatistically significant differences were detected,
a post-hoc pairwise comparison across treatments
was performed using Fisher’s probability of leastsignificant difference (PLSD) test for multiple comparisons. For the comparison among groups, analyses were performed by Tukey-Kramer test on all test
items. However, this study is preliminary. Therefore, distinctions of weight index relating to bodyfat, BMI, waist circumference, hip circumference,
total fat area, subcutaneous fat area and visceral fat
area between the group consumed test food (100,200, and 300 mg) and the group consumed placebo
food were analysed by unpaired t-test.
All statistical analyses were performed using
Statview ver. 5.0 (SAS Institute Japan Ltd., Tokyo,
Japan), and significance was set at p < 0.05.

RESULTS
Exclusion from the Study
One subject in group II withdrew his consent
for participation and was excluded from the study;
however, this was not due to any problem with thetest food.
Subjects who had not eaten home-delivered
meals for more than 10% of all meals were excluded
from the study. The number of subjects was 18, 15,
18, and 18 for groups I, II, III, and IV, respectively.
Subject characteristics are listed in Table 2.
Haematological Examinations and Blood Biochemical Analyses
The results of the haematological examinations
are shown on Table 3. At week 8, the WBC countin group III and the Ht value in group I had changed
significantly from the initial values. However, theWBC count remained within the normal range. Although the average Ht values recorded for group I
at week 8 exceeded the normal values for females,
the values recorded for individual female subjects inthis group were all within the normal range. Thus,
no abnormalities were noted during the haematological examinations (RBC, WBC, Ht, and Hb).
The results of the blood biochemical analyses
are shown in Tables 4 and 5. Although the T-Choand HDL-Cho levels in all the groups had reduced
significantly at week 4 compared to week 0, they
returned to their original values at week 8. The
LDL-Cho levels reduced significantly in groups I
and III at week 4, but returned to their original normal valves at week 8. Although the LDL-Cho levels

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No. 6

Table 3. Hematological Examination
Group
I
II
III
IV

number18
15
18
18

0 week
6.644 ± 1.641
6.693 ± 2.045
6.967 ± 1.304
6.578 ± 1.196

8 week6.600 ± 1.721
6.513 ± 1.904
6.439 ± 1.501∗
6.694 ± 1.555

Red blood cell count(× 106 /µl)

I
II
III
IV

18
15
18
18

4.99 ± 0.38
4.85 ± 0.38
5.00 ± 0.39
4.92 ± 0.44

5.05 ± 0.38
4.94 ± 0.34
5.00 ± 0.39
4.93 ± 0.47

Hemoglobin level(g/dl)

I
II
III
IV

18
15
18
18

15.24 ± 1.23
15.01 ± 1.49
15.16 ± 1.41
14.98 ± 1.22

15.22 ± 1.26
15.14 ± 1.32
15.09 ± 1.40
14.83 ± 1.31

Hematocrit
(%)

I
II
III
IV

18
15
18
18

45.66 ± 3.23
44.62 ± 3.80
45.10 ± 3.77
44.87 ± 3.18

46.65 ± 3.38∗∗
45.79 ± 3.42
45.55 ± 4.11
45.39 ± 3.55

White blood cell count(× 103 /µl)

See Fig. 1 for Groups. Values are means ± S.D. ∗ and ∗∗ indicate significantly different from the value
obtained in week 0 at p < 0.05 and p < 0.01, respectively.

recorded for group III exceeded the normal range
at week 8, these levels were higher than normal,
even at week 0. Therefore, the changes in these levels were not considered abnormal. The leptin levels were significantly lower in group III at both ofweeks 4 and 8, as compared to week 0. Group I
significantly showed low value at weeks 8, as compared to week 0. Since leptin is secreted from fat
tissue, this result may be attributable to a reduction
in body fat. The TB level in group II; GOT and
GPT levels in groups I, II, and III; uric acid levels
in groups I and III; and the ALP level in group III
were all significantly different from the corresponding values at week 0, but remained within the normal range. Although the average γ-GTP values ofgroups I and III were significantly lower than theweek 0 values, they exceeded the normal range forfemales. Nevertheless, the values recorded for individual female subjects in these 2 groups were allwithin the normal range. No significant changes
were noted in the other parameters for any of thegroups.
Urinary Examinations
Sugar was not detected in the urine of any of thesubjects throughout the study period. In addition,
proteins were not detected in the urine of any of thesubjects in groups II and III throughout the study
period. For 1 subject belonging to group I, the status of urinary proteins was detected (±) at week 8;
however, proteins were detected (+) in the urine ofthis subject even at week 0. In addition, in the caseof 1 subject belonging to group IV, the status of urinary proteins was detected (±) at week 8; however,this group was the placebo group. Therefore, we
considered that PFE consumption did not cause anyof the abnormalities detected during the urine examinations.
Blood was detected (+) in the urine of 1 subject
belonging to group III in week 8, but this subject
exhibited bloody urine throughout the study period.
In group II, the status of blood in the urine was assessed as (2+) in 1 subject, (+) in 1 subject, and (±)
in 1 subject at week 8. In the case of the subject forwhom the status was found to be (2+), the blood in
the urine was confirmed to be related to menstruation, according to the recordings in a diary that had
been maintained for this patient. With regard to thesubject for whom the status was assessed as (+), a
status ranging from (±) to (+) was noted throughout
the study period. Further, in the case of the subject for whom the status of blood in the urine wasassessed as ±, a similar status had been noted during prior examinations. Thus, our observations may
have been due to an inherent characteristic of thissubject.
In 1 subject belonging to group IV, the statusof blood in the urine was found to be (2+) at week
8, which was identical to the status found at week
0. Therefore, we consider that our findings may be
attributed to the inherent characteristics of this subject.

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Vol. 57 (2011)

Table 4. Biochemical Examination (Liver and Renal Function)
Group
I
II
III
IV

Number18
15
18
18

Week 07.41 ± 0.36
7.29 ± 0.25
7.41 ± 0.25
7.41 ± 0.26

Week 87.38 ± 0.36
7.25 ± 0.32
7.31 ± 0.33
7.29 ± 0.32

Albumin(g/dl)

I
II
III
IV

18
15
18
18

4.57 ± 0.24
4.50 ± 0.21
4.59 ± 0.17
4.62 ± 0.22

4.62 ±
4.55 ±
4.56 ±
4.61 ±

0.25
0.24
0.18
0.22

Total bilirubin(mg/dl)

I
II
III
IV

18
15
18
18

0.67 ± 0.17
0.72 ± 0.23
0.72 ± 0.35
0.68 ± 0.23

0.68 ±
0.61 ±
0.64 ±
0.65 ±

0.27
0.22∗
0.27
0.22

GOT
(IU/l)

I
II
III
IV

18
15
18
18

26.4 ± 7.8
25.5 ± 13.6
24.0 ± 11.3
24.8 ± 8.4

22.0 ± 5.3∗∗
19.9 ± 7.0∗
20.4 ± 7.6∗
24.9 ± 15.9

GPT
(IU/l)

I
II
III
IV

18
15
18
18

43.7 ± 24.4
37.3 ± 31.1
38.4 ± 26.5
36.2 ± 24.1

35.1 ± 22.2∗
25.9 ± 16.8∗
29.2 ± 19.7∗∗∗
37.2 ± 29.8

ALP
(IU/l)

I
II
III
IV

18
15
18
18

221.1 ± 39.8
221.2 ± 52.1
195.4 ± 47.9
213.4 ± 62.6

226.7 ± 50.4
224.4 ± 55.3
206.6 ± 50.7∗
220.5 ± 65.3

LDH
(IU/l)

I
II
III
IV

18
15
18
18

195.7 ± 34.4
188.0 ± 33.6
177.0 ± 28.1
190.5 ± 27.7

186.4 ± 25.7
186.7 ± 27.7
170.3 ± 24.1
191.6 ± 26.5

γ-GTP
(IU/l)

I
II
III
IV

18
15
18
18

55.6 ± 33.6
51.3 ± 46.4
43.7 ± 37.7
45.4 ± 32.5

42.9 ± 21.7∗∗
41.3 ± 28.1
34.4 ± 24.2∗
48.9 ± 39.7

BUN
(mg/dl)

I
II
III
IV

18
15
18
18

14.6 ± 4.3
12.5 ± 2.2
12.8 ± 2.1
13.1 ± 2.5

12.7 ±
12.1 ±
13.3 ±
12.8 ±

2.5
2.9
3.0
2.5

Creatinine(mg/dl)

I
II
III
IV

18
15
18
18

0.80 ± 0.13
0.81 ± 0.14
0.79 ± 0.12
0.77 ± 0.10

0.79 ±
0.80 ±
0.79 ±
0.78 ±

0.13
0.14
0.11
0.10

Uric acid(mg/dl)

I
II
III
IV

18
15
18
18

6.55 ± 1.70
6.21 ± 1.51
5.98 ± 1.60
6.09 ± 1.29

5.94 ±
6.05 ±
5.68 ±
6.07 ±

1.55∗∗∗
1.09
1.55∗
1.52

CPK
(IU/l)

I
II
III
IV

18
15
18
18

155.3 ± 81.9
118.0 ± 52.4
110.9 ± 43.2
127.4 ± 52.9

Total protein(g/dl)

156.6 ± 101.7
118.4 ± 47.7
104.6 ± 33.6
124.1 ± 74.3

See Fig. 1 for Groups. Values are means ± S.D. ∗ , ∗∗ , and ∗∗∗ indicate significantly different fromthe value obtained in week 0 at p < 0.05, p < 0.01, and p < 0.001, respectively.

527

No. 6

Table 5. Biochemical Examination (Lipid and Glucose Levels)
Group
I
II
III
IV

Number18
15
18
18

Week 0197.2 ± 25.1
202.1 ± 54.3
215.4 ± 29.9
208.6 ± 43.7

Week 4186.8 ± 26.9∗
191.0 ± 45.0∗∗
194.0 ± 23.2∗∗∗
194.1 ± 33.5∗∗

Week 8198.1 ± 27.7
202.3 ± 54.4
208.1 ± 29.9
209.2 ± 30.8

LDL cholesterol(mg/dl)

I
II
III
IV

18
15
18
18

135.7 ± 24.1
139.4 ± 54.4
146.7 ± 27.1
141.0 ± 34.8

126.2 ± 25.9∗∗
127.9 ± 43.9
132.0 ± 18.0∗∗
131.5 ± 31.0

134.0 ± 26.5
133.8 ± 51.6
139.1 ± 25.5
137.1 ± 28.5

HDL cholesterol(mg/dl)

I
II
III
IV

18
15
18
18

48.8 ± 9.4
52.6 ± 8.3
50.2 ± 10.7
49.7 ± 13.1

43.6 ± 6.8∗∗∗
49.5 ± 8.7∗
47.7 ± 9.9∗
45.8 ± 7.9∗

49.1 ± 8.1
52.9 ± 10.2
50.7 ± 11.4
51.2 ± 9.9

RLP cholesterol(mg/dl)

I
II
III
IV

18
15
18
18

6.4 ± 3.2
4.9 ± 2.4
6.8 ± 5.0
7.1 ± 3.6

6.6 ± 3.6
4.9 ± 2.8
5.6 ± 3.3
5.6 ± 2.4

5.1 ± 2.9
4.9 ± 2.1
6.4 ± 3.9
5.8 ± 2.1

Triglycerides(mg/dl)

I
II
III
IV

18
15
18
18

139.0 ± 71.9
104.4 ± 57.3
148.8 ± 84.6
154.5 ± 61.2

160.7 ± 67.2
119.9 ± 63.8
135.4 ± 66.1
138.2 ± 47.2

127.9 ± 71.3
109.6 ± 42.0
137.5 ± 62.2
138.1 ± 44.2

Fasting blood glucose(mg/dl)

I
II
III
IV

18
15
18
18

90.3 ± 5.8
91.1 ± 6.0
89.9 ± 4.8
90.1 ± 4.5

90.7 ± 7.2
91.2 ± 5.9
91.4 ± 8.9
91.3 ± 8.0

91.9 ± 6.4
92.7 ± 8.9
91.1 ± 4.2
89.7 ± 6.6

Insulin(mu/ml)

I
II
III
IV

18
15
18
18

9.2 ± 3.5
7.0 ± 3.0
8.1 ± 3.3
11.6 ± 8.4

9.3 ± 4.2
7.3 ± 3.6
8.2 ± 3.6
11.5 ± 8.7

9.9 ± 5.5
8.1 ± 4.0
9.1 ± 4.5
11.1 ± 5.6

HbA1c
(%)

I
II
III
IV

18
15
18
18

4.81 ± 0.22
4.81 ± 0.20
4.77 ± 0.35
4.82 ± 0.23

—
—
—
—

4.76 ± 0.20
4.83 ± 0.25
4.81 ± 0.32
4.84 ± 0.23

Leptin(ng/dl)

I

16
2
12
3
15
3
15
3

6.9 ± 2.4
14.9 ± 8.1
7.3 ± 4.2
14.9 ± 8.1
6.4 ± 4.6
14.2 ± 5.0
7.0 ± 3.4
14.6 ± 5.7

6.0 ± 2.7
9.9 ± 0.1
4.8 ± 2.0
12.8 ± 6.0
4.7 ± 2.9∗∗
10.7 ± 2.1
6.6 ± 3.9
10.6 ± 2.8

5.8 ± 3.0∗
7.6 ± 0.8
4.9 ± 2.4
12.3 ± 4.2
4.5 ± 2.3∗∗
10.1 ± 3.5
6.2 ± 3.8
10.2 ± 2.7

Total cholesterol(mg/dl)

II
III
IV

male
femalemale
femalemale
femalemale
femaleSee Fig. 1 for Groups. Values are means ± S.D. ∗ , ∗∗ , and ∗∗∗ indicate significantly different from the value obtained in week 0 at p < 0.05,
p < 0.01, and p < 0.001, respectively.

Rational Symptoms
The findings with regard to the rational symptoms are shown in Table 6. Various subjective
symptoms occurred in all groups. However, a doctor
judged there were no relations between those symptoms and the test food.

Physical Examination
Here, we describe the rates of change in theparameters that were assessed during the physicalexaminations. To evaluate the effects of PFE consumption on body fat, we analysed 58 subjects with
BMI values > 24, who were defined as obese ac-528Vol. 57 (2011)

Table 6. Number of Occurrence of Rational Symptoms
Cold
Vomiting
Skin eruption
Menstruation
Abdominal discomfort
Gastric pain
Abdominal pain
Abdominal tension
Constipation
Diarrhoea

Group I
79 (0)
0 (0)
0 (0)
2 (0)
0 (0)
2 (0)
0 (0)
2 (0)
0 (0)
25 (0)

Group II
13 (0)
1 (0)
0 (0)
4 (0)
0 (0)
1 (0)
0 (0)
0 (0)
0 (0)
16 (0)

Group III
14 (0)2 (0)0 (0)21 (0)
0 (0)
0 (0)
1 (0)
1 (0)
0 (0)
20 (0)

Group IV
25 (0)0 (0)0 (0)20 (0)
2 (0)
1 (0)
3 (0)
5 (0)
9 (0)
36 (0)

Total sum of rational symptoms (the number of symptoms confirmed a relation to the test
food by a doctor). See Fig. 1 for Groups.

Table 7. Subject Backgrounds (BMI > 24, Male)

Age (years)
Body weight (kg)BMI (kg/m2 )Waist circumference (cm)Hip circumference (cm)
Total fat area (cm2 )Visceral fat area (cm2 )Subcutaneous fat area (cm2 )Systolic blood pressure (mmHg)
Diastolic blood pressure (mmHg)

Group I (male)(n = 14)40.3 ± 8.6
78.2 ± 7.9
27.5 ± 2.4
97.2 ± 8.6
101.2 ± 5.3
360.6 ± 83.6
138.3 ± 37.1
222.4 ± 68.0
130.0 ± 11.1
81.6 ± 8.0Group II (male)(n = 11)37.1 ± 8.0
78.4 ± 10.2
27.1 ± 3.4
96.1 ± 11.5
99.4 ± 5.8
330.6 ± 145.9
151.3 ± 130.5
179.2 ± 39.1
134.8 ± 12.2
84.5 ± 11.1

Group III (male)(n = 11)37.8 ± 11.1
76.6 ± 8.5
26.5 ± 3.2
97.3 ± 6.2
102.0 ± 3.9
349.5 ± 80.2
130.2 ± 40.5
219.4 ± 69.3
118.5 ± 10.0
75.5 ± 10.4

Group IV (male)(n = 14)34.6 ± 9.0
78.5 ± 8.2
26.8 ± 2.7
97.9 ± 8.5
102.2 ± 4.3
335.5 ± 86.9
131.5 ± 56.4
204.0 ± 54.2
127.6 ± 16.8
81.1 ± 14.0

See Fig. 1 for Groups. Values are means ± S.D.

Table 8. Energy Intake and Amount of Exercise (BMI > 24, Male)

Total energy intake(kcal/day)Number of steps(steps/day)Group I (male)(n = 14)2132.5 ± 268.4

Group II (male)(n = 11)2186.3 ± 279.5

Group III (male)(n = 11)2086.2 ± 207.0

Group IV (male)(n = 14)2065.1 ± 209.7

5436.2 ± 2346.5

7112.0 ± 2531.6

6126.7 ± 1635.4

6832.3 ± 2888.6

See Fig. 1 for Groups. Values are means ± S.D.

cording to the criteria calculated by the analysis ofcovariance (ANCOVA).12) We analysed test itemsof physical examination on only male subjects because there were few female subjects. Thus, 14, 11,
11, and 14 subjects in groups I, II, III, and IV, respectively, were assessed as obese. The characteristics of these subjects are shown in Table 7, and energy intake and amount of exercise received during
PFE intake are shown in Table 8. There were no significant differences among the groups with regard tothese parameters.
The results of the physical examinations areshown in Table 9. At weeks 4 and 8, the BMI andbody weight were significantly reduced in group III
only, and not in any of the other groups. The total
fat area and subcutaneous fat area were significantly
reduced in both groups II and III. At week 4, the hip
circumference was significantly reduced in groups I,
II, and III. The visceral fat area was not significantly
different from that recorded at week 0 in any of thegroups. As a result of comparison among groups by
Tukey-Kramer test, no significant differences werenoted among the groups for any of the abovementioned parameters.

529

No. 6

Table 9. Change Ratio in Physical Examination (BMI > 24, Male)
Group
I
II
III
IV

Number14
11
11
14

Week 01.000 ± 0.000
1.000 ± 0.000
1.000 ± 0.000
1.000 ± 0.000

Week 4
0.992 ± 0.019
0.998 ± 0.020
0.989 ± 0.013∗∗
0.995 ± 0.013

Week 80.987 ± 0.027
0.991 ± 0.023
0.975 ± 0.013∗∗∗
0.993 ± 0.018

BMI(kg/m2 )

I
II
III
IV

14
11
11
14

1.000 ± 0.000
1.000 ± 0.000
1.000 ± 0.000
1.000 ± 0.000

0.992 ± 0.020
0.998 ± 0.021
0.988 ± 0.013∗∗
0.995 ± 0.013

0.987 ± 0.027
0.992 ± 0.023
0.975 ± 0.013∗∗∗
0.993 ± 0.017

Waist circumference
(cm)

I
II
III
IV

14
11
11
14

1.000 ± 0.000
1.000 ± 0.000
1.000 ± 0.000
1.000 ± 0.000

—
—
—
—

0.996 ± 0.024
0.993 ± 0.030
0.976 ± 0.021∗∗
0.990 ± 0.037

Hip circumference
(cm)

I
II
III
IV

14
11
11
14

1.000 ± 0.000
1.000 ± 0.000
1.000 ± 0.000
1.000 ± 0.000

0.981 ± 0.029∗∗
0.984 ± 0.021∗∗
0.971 ± 0.036∗∗
0.989 ± 0.021

0.996 ± 0.023
0.999 ± 0.011
0.984 ± 0.036
1.007 ± 0.033

Total fat area
(cm2 )

I
II
III
IV

14
11
11
14

1.000 ± 0.000
1.000 ± 0.000
1.000 ± 0.000
1.000 ± 0.000

—
—
—
—

0.971 ± 0.082
0.951 ± 0.060∗
0.902 ± 0.070∗∗
0.968 ± 0.105

Subcutaneous fat area
(cm2 )

I
II
III
IV

14
11
11
14

1.000 ± 0.000
1.000 ± 0.000
1.000 ± 0.000
1.000 ± 0.000

—
—
—
—

0.979 ± 0.131
0.928 ± 0.078∗
0.886 ± 0.065∗∗∗
0.963 ± 0.122

Visceral fat area
(cm2 )

I
II
III
IV

14
11
11
14

1.000 ± 0.000
1.000 ± 0.000
1.000 ± 0.000
1.000 ± 0.000

—
—
—
—

0.974 ± 0.166
0.958 ± 0.162
0.950 ± 0.140
1.006 ± 0.211

Systolic blood pressure(mmHg)

I
II
III
IV

14
11
11
14

1.000 ± 0.000
1.000 ± 0.000
1.000 ± 0.000
1.000 ± 0.000

0.949 ± 0.063
0.934 ± 0.056∗∗
0.948 ± 0.082
0.976 ± 0.051

0.966 ± 0.084
0.921 ± 0.066∗∗∗
0.965 ± 0.062
0.961 ± 0.086

Diastolic blood pressure(mmHg)

I
II
III
IV

14
11
11
14

1.000 ± 0.000
1.000 ± 0.000
1.000 ± 0.000
1.000 ± 0.000

0.970 ± 0.115
0.982 ± 0.041
0.898 ± 0.106∗∗
0.952 ± 0.142

0.986 ± 0.133
0.936 ± 0.078∗∗
0.901 ± 0.103∗∗0.930 ± 0.140

Body weight(kg)

See Fig. 1 for Groups. Values are means ± S.D. ∗ , ∗∗ , and ∗∗∗ indicate significantly different from the value obtained in week 0 at
p < 0.05, p < 0.01, and p < 0.001, respectively.

The result of comparison among groups by unpaired t-test was appeared in Fig. 2. Regarding
weight, BMI and total fat area, group III exhibited
significantly lower value than group IV (weight data
is not shown).

DISCUSSION
In Japan, the number of obese persons with BMI
values from 25–30 has increased by 2- to 4-fold overthe last 30 years. Based on BMI values, 18.9% of

the female population and 21.4% of the male population are currently considered as preobese. Thismay be due to the westernization of the food culture
in Japan, which has increased the fat intake to twicethat in the 1960s.
Obesity is the cause of many adult diseases associated with lifestyle habits, including diabetes,
hyperlipidaemia, and hypertension. The type-1
plasminogen activator inhibitor (PAI-1) and agentsthat promote thrombus formation are excessively secreted by the cells of visceral fat tissue. This resultsin adiponectin hyposecretion, and this condition in

530

Vol. 57 (2011)

Fig. 2. Change Ratio in BMI and Total Fat Area (Male)
Result of comparison between group III and group IV.

turn induces insulin resistance, abnormalities in glucose metabolism, hypertension, and arteriosclerosis. Therefore, visceral adiposity is considered adangerous condition and is termed the ‘metabolic
syndrome’. To prevent its development, it is important to improve the status of obesity.
It is well known that catechin,13–15) mannooligosaccharides,16, 17) and coffee polyphenols18)
reduce body fat; however, it is unclear whether these
ingredients improve liver function. The incidenceof nonalcoholic steatohepatitis (NASH) is known to
be strongly associated with obesity. Therefore, theconsumption of ingredients that have effects in both
reducing body fat and improving liver function iseffective for the prevention of this condition.
The puerariae flower is used in Japanese andChinese folk medicine for curing hangovers.1– 3)
Additionally, Niiho et al. confirmed that its consumption reduces GOT or GPT levels in individuals with liver injury induced by CCl4 or a high-fat
diet in animal study.2, 3) Further, Lee et al. have reported the hepatoprotective effects of tectoridin andtectorigenin, which are isoflavone constituents ofpuerariae flos.19) Therefore, it is considered that theconsumption of the puerariae flos may improve liverfunction. Since the ability of PFE to reduce body fat
in vivo has been confirmed, it is expected to be efficient for improving the status of both body fat andliver function.
In this study, we preliminary investigated the effects of PFE on body fat in humans. Among thosesubjects with BMI values of more than 24, signifi-

cant reductions were noted in BMI and body weight
only in group III and in the total fat area and the
subcutaneous fat area in groups II and III (Table 9).
Although the visceral fat area was not significantly
reduced in any of the groups, group III exhibitedthe highest reduction rate. No significant difference
was confirmed on a comparison between placebogroup and PFE group by Tukey-Kramer test. On theother hand, BMI value of male subjects in group III
was significantly low compared with male subjectsof group IV by unpaired t-test (Fig. 2). Althoughmuscle mass was estimated, significant differenceswere not confirmed in any groups by impedance
method (data not shown). In addition, significant
differences were not confirmed on creatinine andred blood cell count that related to amount of water within the body. Therefore, BMI reduction may
reflect the reduction of fat area. In fact, value of total fat area of male subjects in group III tended to be
reduced compared with male subjects of group IV
by unpaired t-test (p < 0.10).
GOT, GPT and γ-GTP as hepatic function markers were shown significantly low value on group I
through group III at 8 weeks (Table 4). As describedabove, hepatoprotective effects of puerariae flos has
been reported with the perception from in vivo.2, 3)
Therefore a possibility is considered that reductionof these makers may be attributable to hepatoprotective effect. This is very interesting.
In this study, we conducted the double-blind
placebo controlled study to examine whether PFE
has anti-obesity effect or not preliminary. As a

531

No. 6

result, BMI value of male subjects in 300 mg/day
group was significantly low compared with malesubjects of placebo group. Consequently, PFE intake at 300 mg/day seems to have the possibility ofanti-obesity effect on male. Since most subjects
were male in this study, we could not confirm theactions on body fat of female. We need to confirm
anti-obesity effect of PFE on female and the optimum dose for administration by a larger scale study.
For the study, enough quantity of sample size should
be ensured and equal numbers of male and female
are required as subjects.

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