Integrative Molecular Medicine

Research Article ISSN: 2056-6360

Enhancement of physical fitness by black ginger extract

rich in polymethoxyflavones: a double-blind randomized
crossover trial
Kazuya Toda, Marina Kohatsu, Shogo Takeda, Shoketsu Hitoe, Norihito Shimizu, and Hiroshi Shimoda*
Research and Development Division, Oryza Oil & Fat Chemical Co., Ltd., 1 Numata, Kitagata-cho, Ichinomiya, Aichi 493-8001, Japan

Abstract
Background: Black ginger (Kaempferia parviflora) contains polymethoxyflavones, which are flavonoids that exhibit various bioactivities including improvements in
muscular metabolism. We examined the effects of black ginger extract (KPE) rich in polymethoxyflavones on physical fitness and fatigue.
Methods: 24 healthy volunteers were recruited. They were randomly divided into two groups: group A received KPE (30 mg/day) and then a placebo while group B
received a placebo and then KPE in a crossover trial. Each volunteer took one capsule containing KPE or the placebo once a day for 4 weeks. A physical fitness test
(PFT), questionnaire, and blood test were performed at 0, 4, 7, and 11 weeks (wash-out term for 3 weeks).
Results: After a 4-week ingestion period, improvements in left hand grip strength (2.80 vs 0.03 kg), performance in the 30-second chair stand test (6.27 vs 1.71 times),
5-m tandem walking test (-3.17 vs -0.87 sec), and cycle ergometer test (8.54 vs 1.13 kcal) were significantly greater in the KPE group than in the placebo group. In
a fatigue of subjects without an exercise habit, the mean reductions after ingestion of KPE in the daily VAS fatigue score (-9.87 vs +1.52%), post PFT VAS fatigue
score (-10.2 vs -0.91%), and chronic fatigue syndrome (CFS) score (-3.93 vs -2.47) were greater than the placebo group.
Conclusions: The ingestion of KPE was found to enhanced physical fitness, namely, grip strength, leg strength, balance, endurance, and locomotor activity. Furthermore,
KPE intake slightly improved fatigue at the conventional state and post PFT state as well as CFS scores in subjects without an exercise habit

List of abbreviations improve muscular metabolism and reduce oxidation stress.

AICAR: 5-aminoimidazole-4-carboxyamide ribonucleotide; Black ginger, the rhizome of Kaempferia parviflora (Zingiberaceae)
AMPK : AMP-activated protein kinase; KPE: black ginger (Kaempferia has traditionally been used in folk medicines and nourishing foods
parviflora) extract; BCAA: branched chain amino acids; CFS: in Thailand, and contains polymethoxyflavones (PMFs), which are
chronic fatigue syndrome; MCV: mean cell volume; MCH: mean flavonoids that exhibit various bioactivities (i.e. anti-inflammatory,
corpuscular hemoglobin; MCHC: mean corpuscular hemoglobin antioxidant [18-21], anticancer [22,23], muscular metabolism-
concentration; PDE:phosphodiesterase; PFT: physical fitness test; enhancing [24], anti-photoaging [25], phosphodiesterase (PDE)5
PMFs: polymethoxyflavones; QOL: quality of life; TG: triglyceride; inhibitory [26], anti-cardiovascular disease [27], and viral protease
VAS: visual analog scale inhibitory [28] activities). A large number of studies have investigated
the bioactivities of black ginger extract (KPE), which is rich in PMFs
Backgrounds [24,29-37]. We also reported that PMFs in KPE improved muscular
Locomotive dysfunctions, which are related to muscles, bones, and metabolism through AMP-activated protein kinase (AMPK) activation
joints, account for some of the main factors impairing the quality of in myocytes [24]. Furthermore, clinical trials have been conducted on
life (QOL) of individuals. Muscular dysfunctions including sarcopenia the effects of KPE physical performance in athletes and elderly subjects
syndrome may trigger locomotive dysfunctions and are caused by [36,37].
declines in muscular mass and metabolism, which are induced by However, clinical reports have yet to be performed on healthy
aging or insufficient daily exercise [1-5]. Therefore, continual exercise adults aged between 20 and 65 years old. In addition, the effects of KPE
is necessary in order to prevent locomotive dysfunctions [6-9]. In
addition, supportive strategies to enhance exercise performance such
as ingestion of nutrients may be effective. Amino acids and peptide
derivatives such as branched chain amino acids (BCAA) and imidazole Correspondence to: Hiroshi Shimoda, Research & Development Division, Oryza
peptides are often used in dietary supplements prescribed to improve Oil & Fat Chemical Co., Ltd., 1 Numata, Kitagata-cho, Ichinomiya, Aichi 493-
muscle function. These nutritional ingredients were previously reported 8001, Japan, Tel.: +81-586-86-5141; Fax: +81-586-86-6191; E-mail: kaihatsu@
to be effective in clinical trials [10]. One of the mechanisms responsible mri.biglobe.ne.jp
is an increase in the mass of skeletal muscle through protein synthesis. Key words: randomized, double-blind, placebo-controlled crossover trial,
On the other hand, co-enzyme Q10 [11-14], L-carnitine [15,16], and kaempferia parviflora, polymethoxyflavone,exercise,physical fitness, fatigue
polyphenol [17] have been reported as ingredients that effectively Received: April 01, 2016; Accepted: April 19, 2016; Published: April 23, 2016

Integr Mol Med, 2016 doi: 10.15761/IMM.1000215 Volume 3(2): 628-634

 Toda K (2016) Enhancement of physical fitness by black ginger extract rich in polymethoxyflavones: a double-blind randomized crossover trial

on subjects without an exercise habit or on post-exercise fatigue have Table 1. Subject backgrounds.
not yet been investigated. Therefore, we examined the effects of KPE on General characteristics Group A Group B Total
physical fitness and fatigue in healthy volunteers. All subjects
Male 8 8 16
Methods Female 4 4 8
Total 12 12 24
Test design
Ages (year) 35.4 ± 3.5 35.0 ± 2.9 35.2 ± 2.2
This study was performed as a randomized double-blind placebo Subjects with an exercise habit
controlled crossover trial guided by the directions of the 6th revision of Male 5 3 8
the Declaration of Helsinki (2008) and guidelines of the consolidated Female 0 1 1
standards of reporting (CONSORT 2010 Statement, Japan). In Total 5 4 9
addition, the study was registered in the University Hospital Medical Ages (year) 33.6 ± 3.2 37.8 ± 3.3 35.4 ± 2.2
Information Network (UMIN, ID: UMIN000021051). The protocol was Subjects without an exercise habit
approved by the Ethics Committee in Oryza Oil & Fat Chemical Co., Male 3 5 8
Ltd. (October 1, 2015, Approval No.20151001) without contribution Female 4 3 7
to the trial. The study was performed based on the schedule described Total 7 8 15
in Figure 1. Ages (year) 35.4 ± 3.5 35.0 ± 2.9 35.1 ± 2.3
First sample KPE Placebo
Subjects Second sample Placebo KPE
Data for ages were presented as the mean ± S. E.
Subjects were recruited as healthy volunteers from Oryza Oil &
Fat Chemical Co., Ltd., and were aged between 20 and 65 years old.
Key exclusion criteria were 1) subjects receiving medication or having age by a third member. Information on the assignment was concealed
anamnesis of serious diseases requiring medication, 2) subjects with by the third member until fixation of the data. Subject backgrounds in
chronic diseases including asthma and 3) subjects who were allergic to each group were shown in Table 1.
the test sample.
Test samples
The essential number of subjects was determined as 23 by G*Power,
KPE was prepared according to our previously reported method
software for a power analysis. Twenty four subjects (male: 16, female:
[24]. KPE was obtained from the dried rhizomes of black ginger by
8) received full explanations on the purpose and method of the study,
extracting with aqueous ethanol (yield 15.9%). KPE was mixed with
and then participated in the trial with their consent. Male and female
modified starch at a ratio of 3:7 (KPE: modified starch) and then
subjects were randomly distributed into groups A and B in order of
powdered by spray drying. Powdered KPE (100 mg) was packed
into a capsule. The contents of total PMFs in the KPE capsule were
determined by reverse-phase HPLC using a Prominence HPLC system
(Shimadzu, Kyoto, Japan) equipped with a photodiode array detector
(Model SPD-M20A) and Develosil RPAQUEOUS-AR-5 column (4.6
× 150 mm, 5-µm particle size, Nomura Chemical Co., Ltd., Japan).
The mobile phase was a binary gradient and consisted of a mixture of
acetonitrile, water, and acetic acid (35: 62.5: 2.5, v/v) as solvent A and a
mixture of acetonitrile and acetic acid (97.5: 2.5, v/v) as solvent B. The
flow rate was fixed at 1.0 mL/min and the column temperature was set at 35
°C. Gradient conditions were as follows: 0–20 min (solvent A: 99–1%). UV
detection at 263 nm was used. The contents of the determined PMFs were
5-hydroxy-3,7-dimethoxyflavone (0.66%), 5-hydroxy-7-methoxyflavone
(0.52%), 5-hydroxy-3,7,4’-trimethoxyflavone (0.81%), 5-hydroxy-3,7,3’,4’-
tetramethoxyflavone (0.30%), 3,5,7,3’,4’-pentamethoxyflavone (2.94%),
5,7,4’-trimethoxyflavone (3.14%), 3,5,7,4’-tetramethoxyflavone (1.75%), and
5,7-dimethoxyflavone (2.50%), respectively. In the placebo, modified
starch (100 mg) was packed into a capsule. Subjects took one KPE or
placebo capsule once a day for 4 weeks.
Physical fitness test (PFT)
The hand grip strength test and 30-second chair stand test were
performed according to the methods described in a previous study
[37]. In the 5-m tandem walking test, the time to walk 5 m with tandem
steps was measured. In the cycle ergometer test, energy consumption
(kcal) was measured using an exercise bike (aerobike EZ101, Konami
Sports Club Co. Ltd., Tokyo, Japan). Each subject determined the pedal
load by themselves, peddled at more than 60 rpm for 10 min, and then
peddled with full power for 10 seconds every 2 minutes (a total of 5
times). Locomotor activity (energy consumption: kcal) depended on
Figure 1. Schedule for the study.

Integr Mol Med, 2016 doi: 10.15761/IMM.1000215 Volume 3(2): 628-634

 Toda K (2016) Enhancement of physical fitness by black ginger extract rich in polymethoxyflavones: a double-blind randomized crossover trial

the load and the peddling times for 10 min. Data analysis
Questionnaire Data are presented as the mean ± S.E. In statistical comparisons of
PFT with the placebo, questionnaire, and blood test, a paired t-test or
Fatigue was evaluated by a visual analog scale (VAS) fatigue score
Mann-Whitney’s U test was performed.
and chronic fatigue syndrome (CFS) score. The evaluation of VAS was
performed in the conventional state and post PFT state. The CFS score Results
[38] was determined using the questionnaire in Table 2.
Physical fitness and endurance (all subjects)
Blood test
In order to evaluate the effects of KPE on physical fitness and
After PFT and the questionnaire, blood was collected from endurance, PFT consisting of a hand grip strength test, 30-second
subjects, and the following parameters were analyzed: total bilirubin, chair stand test, 5-m tandem walking test, and cycle ergometer test,
total protein, albumin, A/G ratio, AST, ALT, ALP, LDH, γGTP, CPK, was performed. In Table 3, grip strength after a 4-week treatment with
HDL-cholesterol, LDL-cholesterol, total cholesterol, triglyceride (TG), KPE significantly increased (right: +2.2 kg, P < 0.05, left: +2.8 kg, P
phospholipids, free fatty acids, Na, Cl, K, urea nitrogen, creatinine, < 0.01) from baseline. Significant improvements were also observed
uric acid, glucose, ketone bodies, HbA1c, and cortisol. In addition, in the 30-second chair stand test (+6.3 times, P < 0.01), 5-m tandem
the number of leukocytes and red blood cells, hemoglobin level, walking test (-3.2 sec, P < 0.05), and cycle ergometer test (+8.6 kcal, P
hematocrit, mean cell volume (MCV), mean corpuscular hemoglobin < 0.01). The grip strength of right hand was significantly higher after
(MCH), mean corpuscular hemoglobin concentration (MCHC), and the ingestion of KPE (44.6 kg, P < 0.05) than after that of the placebo
platelet count were determined. (43.0 kg).
Net changes in the grip strength of left hand (+2.8 vs +0.0 kg, P <
0.05), 30-second chair stand test (+6.3 vs +1.7 times, P < 0.05), 5-m
Table 2. Questionnaire for CFS. tandem walking test (-3.2 vs -0.9 sec, P < 0.01), and cycle ergometer test
Questionnaire (+8.5 vs +1.1 kcal, P < 0.01) were significantly greater after the intake of
1) Do you have problems with tiredness? KPE than the values after the placebo ingestion. These results indicate
2) Do you need to rest more? that KPE possesses the ability to enhance physical fitness, namely,
3) Do you feel sleepy or drowsy? grip strength, leg muscle strength, balance, endurance, and locomotor
4) Do you have problems starting things? activity (Table 3).
5) Do you start things without difficulty, but get weak as you continue?
6) Are you lacking in energy?
Physical fitness and endurance (subjects with or without an
7) Do you have less strength in your muscles? exercise habit)
8) Do you feel weak?
In order to evaluate the influence of an exercise habit, data were
9) Do you have difficulty concentrating?
recalculated by a differential analysis with or without an exercise habit.
10) Do you have problems thinking clearly?
An exercise habit was defined as exercise performed once or more a
11) Do you make slips of the tongue when speaking?
week. In subjects with an exercise habit (Table 4, upper part), the grip
12) Do you find it more difficult to find the correct word?
strength of the right hand (+2.2 kg), 30-second chair stand test (+6.9
13) How is your memory?
times), 5-m tandem walking test (-1.7 sec), and cycle ergometer test
14) Have you lost interest in the things you used to do?
(+9.5 kcal) were significantly improved by KPE (P < 0.05). However,
Answer
these improvements were not significantly different from those
1) Better than usual (0 point)
2) No more than usual (1 point)
observed in placebo group.
3) Worse than usual (2 points) In subjects without an exercise habit (Table 4, lower part), the
4) Much worse than usual (3 points) grip strength of left hand (+3.1 kg, P < 0.05), 30-second chair stand
This questionnaire was previously described by Chalder et al. [38]. Subjects scored “better test (+5.9 times, P < 0.05), and cycle ergometer test (+7.9 kcal, P <
than usual” (0 point), “no more than usual” (1 point), “worse than usual” (2 points), and
0.01) were significantly improved by the ingestion of KPE. On the
“much worse than usual” (3 points) for each question. The total number of points was
defined as the CFS score.

Table 3. Effects of KPE on physical fitness and fatigue (part 1).

Measured parameters Placebo KPE

Before After Net change (∆) Before After Net change (∆)
All subjects
Tiredness without exercise (%) 34.6 ± 4.3 32.0 ± 3.5 − 2.57 ± 4.5 34.5 ± 4.0 29.4 ± 4.3 − 5.14 ± 4.2
Grip strength (R) (kg) 42.8 ± 2.5 43.0 ± 2.5 0.21 ± 0.7 42.4 ± 2.4 44.6 ± 2.6 *,† 2.17 ± 0.9
Grip strength (L) (kg) 40.1 ± 2.5 40.1 ± 2.5 0.03 ± 0.8 38.9 ± 2.2 41.7 ± 2.2 †† 2.80 ± 0.8 *
30-second chair stand test (sec) 25.3 ± 2.1 27.0 ± 1.8 1.71 ± 0.9 21.4 ± 1.7 27.6 ± 1.7 †† 6.27 ± 1.7 *
5-m tandem walking test (sec) 12.2 ± 0.8 11.4 ± 0.9 − 0.87 ± 0.5 13.8 ± 1.3 10.6 ± 0.9 † − 3.17 ± 1.3 **
Cycle ergometer test (kcal) 47.4 ± 4.5 48.6 ± 4.3 1.13 ± 1.4 44.3 ± 4.0 52.9 ± 4.7 †† 8.54 ± 1.9 **
Tiredness after this test (%) 56.7 ± 4.5 51.0 ± 4.9 − 5.08 ± 4.3 52.7 ± 5.4 46.8 ± 3.9 − 5.87 ± 4.3
Chronic fatigue syndrome score 16.1 ± 1.2 13.7 ± 1.1 − 2.38 ± 1.1 15.7 ± 1.3 13.0 ± 1.2 − 2.75 ± 1.2

Data were presented as the mean ± S.E (n = 24)

Significant differences from the placebo were indicated as *: P<0.05, **: P<0.01, and those from the baseline were indicated as †: P<0.05, ††: P<0.01 (paired t-test).

Integr Mol Med, 2016 doi: 10.15761/IMM.1000215 Volume 3(2): 628-634

 Toda K (2016) Enhancement of physical fitness by black ginger extract rich in polymethoxyflavones: a double-blind randomized crossover trial

Table 4. Effects of KPE on physical fitness and fatigue (part 2).

Measured parameters Placebo KPE

Before After Net change (∆) Before After Net change (∆)
Subjects with an exercise habit
Tiredness without exercise (%) 34.3 ± 4.0 25.0 ± 2.1 − 9.38 ± 3.8 30.1 ± 2.6 32.8 ± 3.9 − 2.74 ± 2.6
Grip strength (R) (kg) 50.0 ± 2.1 50.1 ± 1.6 0.40 ± 0.8 50.0 ± 1.9 52.2 ± 1.9 † 2.21 ± 0.5
Grip strength (L) (kg) 47.8 ± 1.9 47.2 ± 1.6 − 0.60 ± 1.1 46.2 ± 1.6 48.6 ± 1.6 2.39 ± 0.8
30-second chair stand test (sec) 29.9 ± 2.8 31.7 ± 1.9 1.78 ± 1.3 24.2 ± 1.7 31.1 ± 1.4 † 6.89 ± 1.7
5-m tandem walking test (sec) 10.6 ± 0.6 9.31 ± 0.4 − 1.32 ± 0.4 10.2 ± 0.4 8.47 ± 0.3 † − 1.68 ± 0.3
Cycle ergometer test (kcal) 62.9 ± 4.4 65.2 ± 4.0 2.27 ± 1.3 57.7 ± 3.3 67.3 ± 3.8 † 9.52 ± 1.9
Tiredness after this test (%) 53.2 ± 4.9 41.1 ± 3.7 − 12.1 ± 2.7 36.2 ± 3.8 37.6 ± 4.4 1.37 ± 3.1
Chronic fatigue syndrome score 13.6 ± 1.3 11.3 ± 0.4 − 2.22 ± 1.4 12.8 ± 0.8 12.0 ± 0.9 − 0.78 ± 0.6
Subjects without an exercise habit
Tiredness without exercise (%) 34.8 ± 4.6 36.3 ± 3.9 1.52 ± 4.8 37.1 ± 4.7 27.3 ± 4.6 − 9.87 ± 4.7
Grip strength (R) (kg) 38.7 ± 2.5 38.8 ± 2.6 0.11 ± 0.7 37.9 ± 2.3 40.0 ± 2.6 2.15 ± 1.0
Grip strength (L) (kg) 35.5 ± 2.3 35.9 ± 2.5 0.40 ± 0.5 34.5 ± 2.0 37.6 ± 2.1 † 3.05 ± 0.9
30-second chair stand test (sec) 22.5 ± 1.3 24.2 ± 1.6 † 1.67 ± 0.5 19.6 ± 1.5 25.5 ± 1.8 † 5.90 ± 1.8
5-m tandem walking test (sec) 13.2 ± 0.9 12.6 ± 1.1 − 0.60 ± 0.5 15.9 ± 1.4 11.9 ± 1.1 − 4.06 ± 1.6
Cycle ergometer test (kcal) 38.1 ± 3.5 38.6 ± 3.0 0.44 ± 1.4 36.3 ± 3.5 44.2 ± 4.3†† 7.95 ± 1.9 *
Tiredness after this test (%) 57.8 ± 4.3 56.9 ± 5.2 − 0.91 ± 4.9 62.5 ± 5.3 52.3 ± 3.2 − 10.2 ± 4.7
Chronic fatigue syndrome score 17.6 ± 1.1 15.1 ± 1.3 − 2.47 ± 1.0 17.5 ± 1.4 13.5 ± 1.3 − 3.93 ± 1.4

Data were presented as the mean ± S.E (subjects with an exercise habit: n = 9, subjects without an exercise habit: n = 15).
Significant differences from the placebo were indicated as *: P<0.05, and those from the baseline were indicated as †: P<0.05, ††: P<0.01 (paired t-test).
Table 5. Effects of KPE on blood parameters (part 1).

Measured Unit Normal range Placebo KPE

parameters
Before After Net change (∆) Before After Net change (∆)
Total bilirubin mg/dL 0.2 - 1.2 0.60 ± 0.05 0.65 ± 0.06 0.05 ± 0.05 0.65 ± 0.06 0.60 ± 0.05 - 0.05 ± 0.05
Total protein g/dL 6.5 - 8.3 7.91 ± 0.14 7.86 ± 0.13 - 0.05 ± 0.06 7.72 ± 0.10 7.87 ± 0.10 †† 0.15 ± 0.05 *
Albumin g /dL 3.8 - 5.3 5.00 ± 0.09 4.96 ± 0.10 - 0.03 ± 0.04 4.97 ± 0.08 4.97 ± 0.08 0.00 ± 0.04
A/G ratio 1.1 - 2.3 1.76 ± 0.06 1.75 ± 0.07 0.00 ± 0.03 1.83 ± 0.06 1.74 ± 0.06 † - 0.09 ± 0.03
AST U/L 8 - 38 20.8 ± 0.99 21.8 ± 1.19 1.05 ± 0.64 21.2 ± 1.13 21.0 ± 0.96 - 0.23 ± 0.81
ALT U/L 4 - 43 19.1 ± 1.70 22.1 ± 2.48 † 2.91 ± 1.22 20.7 ± 2.95 21.6 ± 1.83 0.91 ± 2.42
ALP U/L 110 - 354  214 ± 10.3 209 ± 10.5 - 5.73 ± 4.35 210 ± 11.5 208 ± 8.86 - 2.00 ± 6.61
LDH U/L 121 - 245 181 ± 5.77 185 ± 7.53 3.68 ± 4.24 181 ± 6.18 182 ± 6.45 1.18 ± 2.85
γGTP U/L 0 - 86 24.4 ± 2.64 25.4 ± 3.19 1.00 ± 1.08 22.6 ± 2.14 25.8 ± 2.66 † 3.14 ± 1.11
CPK U/L 38 - 196 156 ± 19.1 166 ± 18.5 11.2 ± 11.8 150 ± 15.3 165 ± 21.6 14.3 ± 13.9
LDL-cholesterol mg/dL 70 - 139 120 ± 5.93 121 ± 6.55 1.68 ± 3.03 117 ± 6.27 122 ± 5.23 5.45 ± 2.83
Total cholesterol mg/dL 130 - 219 209 ± 7.23 209 ± 7.60 - 0.27 ± 3.73 205 ± 7.70 211 ± 6.41 5.82 ± 3.87
Triglyceride mg/dL 30 - 149 116 ± 14.3 111 ± 11.7 - 5.14 ± 10.2 119 ± 17.3 116 ± 13.2 - 3.18 ± 17.6
Phospholipids mg/dL 150 - 260 233 ± 7.98 235 ± 7.68 1.73 ± 4.54 231 ± 8.95 239 ± 7.09 8.05 ± 4.20
Free fatty acids mEq/L 0.13 - 0.77 0.45 ± 0.05 0.44 ± 0.05 0.00 ± 0.07 0.48 ± 0.06 0.46 ± 0.07 - 0.03 ± 0.06
HDL-cholesterol mg/dL 40 - 77 68.4 ± 3.65 66.0 ± 2.90 - 2.45 ± 1.81 67.8 ± 3.32 67.6 ± 3.16 - 0.18 ± 1.43
Data were presented as the mean ± S.E (n = 22).

other hand, only performance in the 30-second chair stand test was by ingestion of KPE or the placebo for 4 weeks (mean reduction: KPE
significantly improved (+1.67 times, P < 0.05) after the intake of the -5.14% vs placebo -2.57%). These results were similar to the post PFT
placebo than before its ingestion. A significant difference (P < 0.05) was VAS fatigue scores (mean reduction: KPE -5.87% vs placebo -5.08%)
observed in the net change in cycle ergometer test between the placebo and CFS scores (mean reduction: KPE -2.75 vs placebo -2.38) (Table 3).
group and the KPE group (7.95 vs 0.44 kcal), and this was not detected
Data for subjects without an exercise habit were re-analyzed. The
in subjects without an exercise habit. However, in a differential analysis
results obtained showed that mean reductions after ingestion of KPE
with or without an exercise habit, the average reductions observed in
in the daily VAS fatigue score (-9.87 vs +1.52%), post PFT VAS fatigue
subjects with and without an exercise habit were not significant for any
score (-10.2 vs -0.91%), and CFS score (-3.93 vs -2.47) were greater
parameter tested. Therefore, these results suggest that an exercise habit
than the placebo group (Table 4, lower part). Moreover, in the mean
is not a key factor for KPE-induced enhancements in physical fitness.
reductions observed in the CFS score, the treatment with KPE led
Fatigue in conventional and post-exercise states to slightly better scores in subjects without an exercise habit than in
those with an exercise habit (-3.93 vs -0.78, P=0.10). However, this
Daily and post PFT VAS fatigue scores and CFS score were was not the case in those receiving the placebo (-2.47 vs -2.22, P=0.93).
measured to evaluate the effects of KPE on fatigue. In all subjects, the Therefore, these results suggest that KPE improves fatigue in subjects
daily VAS fatigue score at the conventional state was slightly decreased

Integr Mol Med, 2016 doi: 10.15761/IMM.1000215 Volume 3(2): 628-634

 Toda K (2016) Enhancement of physical fitness by black ginger extract rich in polymethoxyflavones: a double-blind randomized crossover trial

without an exercise habit only. In PFT, net changes in the grip strength of left hand (+2.8 vs +0.0
kg, P < 0.05), 30-second chair stand test (+6.3 vs +1.7 times, P < 0.05),
Blood parameters 5-m tandem walking test (-3.2 vs -0.9 sec, P < 0.01), and cycle ergometer
Blood parameters were analyzed before and after the ingestion of test (+8.5 vs +1.1 kcal, P < 0.01) were significantly better following the
the test samples. Statistical evaluations were performed on 22 subjects intake of KPE than that of the placebo (Table 1). Therefore, KPE was
because blood samples were not collected from two subjects (Tables 5 found to enhance physical fitness, namely, grip strength, leg strength,
and 6). The results obtained showed that ingestion of KPE significantly balance, endurance, and locomotor activity (Table 3).
increased total protein (+0.15 g/dL), γGTP (+3.14 U/L), number of On the other hand, KPE did not significantly change VAS fatigue
leukocytes (+6.95×102 cells/µL), and hematocrit (+0.83%) compared to scores in daily and post PFT and CFS scores compared with the placebo
the values before ingestion, while the A/G ratio (-0.09), MCH (-0.22 (Table 3). However, in subjects without an exercise habit, KPE slightly
pg), and MCHC (-0.40%) were lower compared to the values before improved these fatigue scores compared to the values in the placebo
the ingestion. On the other hand, ingestion of the placebo significantly group following a 4-week ingestion period (Table 4, lower part).
increased ALT (+2.91 U/L), Cl (+0.64 mEq/L), and ketone body (+9.73 Muscular mass and metabolism may be reduced in the skeletal muscle
µmol/L) levels and decreased MCH (-0.27 pg) and HbA1c (-0.11%). of subjects without an exercise habit. KPE was suggested to improve
Significant changes in total protein levels, hematocrit, and HbA1c were these decreases. We previously confirmed that PMFs, particularly
observed in the values after ingestion of placebo and KPE. However, 5,7-dimethoxyflavone in KPE, improved metabolism in muscle cells
these blood parameter changes were within normal ranges. by activating AMPK [24]. AMPK is known to play a critical role in
Discussion the regulation of energy homeostasis and is related to physical fitness,
endurance, and fatigue [41-48]. For example, 5-aminoimidazole-
In the present study, PFT (hand grip strength test, 30-second 4-carboxyamide ribonucleotide (AICAR), an AMPK agonist, was
chair stand test, 5-m tandem walking test, and cycle ergometer test), reported to increase running endurance by 44% and decrease body fat
a questionnaire about daily and post-exercise fatigue, and blood tests in mice following its oral administration for 4 weeks [48]. Therefore,
were performed to investigate the safety and effectiveness of KPE on the activation of AMPK by KPE may contribute to these activities
physical fitness and fatigue. The results of the blood test showed that including improvements in physical fitness performance.
KPE did not induce abnormal changes in blood parameters compared
with those by the placebo, and the significant changes observed were Aging has also been reported to deteriorate muscular mass and
within normal ranges (Tables 5 and 6). Therefore, the safety of KPE was metabolism [1-5]. A clinical report demonstrated that KPE effectively
clarified in this trial condition. increased physical fitness in elderly subjects aged 60 or more [37].
Therefore, KPE may have more prominent effects on physical fitness
Among blood markers related to exercise in the placebo group, and fatigue in elderly people and individuals lacking an exercise habit,
ALT (+2.91 U/L) and ketone body levels (+9.73 µmol/L), which among whom muscular metabolism has declined. A larger scale clinical
increase during skeletal muscle injury [39] or after exercise [40], study of KPE in elderly subjects or subjects without an exercise habit
were significantly increased in the placebo group, but not in the KPE needs to be performed to clarify these hypotheses. In the future, KPE
group (+0.91 U/L and +2.95 µmol/L). Therefore, KPE was suggested rich in PMFs may be utilized to enhance fitness performance and
to decrease the physical load of exercise in order to maintain muscle prevent locomotive dysfunctions.
condition.
Conclusion

Table 6. Effects of KPE on blood parameters (part 2).

Measured parameters Unit Normal range Placebo KPE
Before After Net change (∆) Before After Net change (∆)
Na mEq/L 135 - 150 143 ± 0.45 143 ± 0.37 0.00 ± 0.33 143 ± 0.35 142 ± 0.38 - 0.23 ± 0.32
Cl mEq/L 98 - 110 102 ± 0.39 103 ± 0.32 † 0.64 ± 0.29 103 ± 0.50 102 ± 0.44 - 0.18 ± 0.35
K mEq/L 3.5 - 5.3 4.12 ± 0.08 4.16 ± 0.06 0.04 ± 0.06 4.11 ± 0.05 4.19 ± 0.04 0.08 ± 0.05
Urea nitrogen mg/dL 8.0 - 22.0 13.2 ± 0.82 13.2 ± 0.89 0.05 ± 0.55 13.2 ± 0.72 13.6 ± 0.86 0.40 ± 0.66
Creatinine mg/dL 0.61 - 1.04 0.74 ± 0.04 0.74 ± 0.04 0.00 ± 0.02 0.73 ± 0.03 0.72 ± 0.03 0.00 ± 0.01
Uric acid mg/dL 3.6 - 7.0 5.10 ± 0.28 5.21 ± 0.26 0.10 ± 0.12 5.15 ± 0.28 5.18 ± 0.28 0.03 ± 0.10
Glucose mg/dL 60 - 109 88.6 ± 3.36 87.7 ± 3.16 - 0.86 ± 3.14 85.5 ± 3.60 81.9 ± 3.59 - 3.59 ± 2.17
Ketone bodies μmol/L 0 - 74 25.2 ± 3.13 34.9 ± 3.20 † 9.73 ± 3.72 29.8 ± 2.89 32.8 ± 3.14 2.95 ± 3.59
Leukocyte × 102/μL 39 - 98 80.6 ± 4.10 75.8 ± 4.27 - 4.77 ± 4.47 73.6 ± 3.51 80.5 ± 3.74 † 6.95 ± 2.63
Red blood cell × 102/μL 427 - 570 493 ± 8.82 492 ± 9.46 - 0.32 ± 3.73 490 ± 9.42 497 ± 9.24 6.77 ± 3.29
Hemoglobin g/dL 13.5 - 17.6 15.0 ± 0.33 14.8 ± 0.36 - 0.15 ± 0.13 14.9 ± 0.32 15.0 ± 0.32 0.09 ± 0.11
Hematocrit % 39.8 - 51.8 45.9 ± 0.93 45.6 ± 0.96 - 0.26 ± 0.31 45.3 ± 0.90 46.2 ± 0.95 † 0.83 ± 0.36 *
MCV fL 82.7 - 101.6 93.1 ± 0.76 92.6 ± 0.78 - 0.44 ± 0.43 92.5 ± 0.76 92.9 ± 0.80 0.39 ± 0.30
MCH pg 28.0 - 34.6 30.4 ± 0.29 30.1 ± 0.31 †† - 0.27 ± 0.06 30.4 ± 0.25 30.2 ± 0.29 † - 0.22 ± 0.09
MCHC % 31.6 - 36.6 32.7 ± 0.22 32.5 ± 0.19 - 0.15 ± 0.16 32.9 ± 0.21 32.5 ± 0.23 †† - 0.40 ± 0.13
Platelet × 104/μL 13.1 - 36.2 27.7 ± 1.25 28.3 ± 1.41 0.62 ± 0.59 27.7 ± 1.13 28.7 ± 1.32 0.97 ± 0.55
HbA1c NGSP % 4.6 - 6.2 5.35 ± 0.06 5.24 ± 0.05 †† - 0.11 ± 0.02 5.29 ± 0.05 5.28 ± 0.05 - 0.01 ± 0.03 *
Cortisol μg/dL 6.2 - 19.4 12.8 ± 1.42 13.03 ± 1.04 0.20 ± 1.05 11.8 ± 1.06 11.7 ± 0.87 - 0.10 ± 0.65
Data were presented as the mean ± S.E (n = 22).
Significant differences from the placebo were indicated as *: P<0.05, and those from the baseline were indicated as †: P<0.05, ††: P<0.01 (paired t-test).

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 Toda K (2016) Enhancement of physical fitness by black ginger extract rich in polymethoxyflavones: a double-blind randomized crossover trial

We herein demonstrated that a 4-week treatment with KPE rich 19. Seito LN, Sforcin JM, Bastos JK, Di Stasi LC (2015) Zeyheria montana Mart.
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in PMFs enhanced performance in the hand grip test, 30-second beneficial effects on intestinal inflammation. J Pharm Pharmacol 67: 597-604.
chair stand test, 5-m tandem walking test, and cycle ergometer test. [Crossref]
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22. Kim M, Kim N, Han J (2014) Metabolism of Kaempferia parviflora

Conflicts of Interest polymethoxyflavones by human intestinal bacterium Bautia sp. MRG-PMF1. J Agric
Food Chem 62: 12377-12383. [Crossref]
All authors related to this study are employees of Oryza Oil & Fat
Chemical Co., Ltd. (Aichi, Japan). The authors declare no conflict of 23. Tsuji PA, Winn RN, Walle T (2006) Accumulation and metabolism of the anticancer
flavonoid 5,7-dimethoxyflavone compared to its unmethylated analog chrysin in the
interest associated with this manuscript. Atlantic killifish. Chem Biol Interact 164: 85-92. [Crossref]
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Copyright: ©2016 Toda K. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use,
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Integr Mol Med, 2016 doi: 10.15761/IMM.1000215 Volume 3(2): 628-634