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Astragalus membranaceus (AM) is a popular "Qi-tonifying" herb with a long history of use as a Traditional Chinese Medicine with multiple biological functions. However, evidence for the effects of AM on exercise performance and physical fatigue is limited. We evaluated the potential beneficial effects of AM on ergogenic and anti-fatigue functions following physiological challenge. Male ICR strain mice were randomly assigned to four groups (n = 10 per group) for treatment: (1) sedentary control and vehicle treatment (vehicle control); (2) exercise training with vehicle treatment (exercise control); and (3) exercise training with AM treatment at 0.615 g/kg/day (Ex-AM1) or (4) 3.075 g/kg/day (Ex-AM5). Both the vehicle and AM were orally administered for 6 weeks. Exercise performance and anti-fatigue function were evaluated by forelimb grip strength, exhaustive swimming time, and levels of serum lactate, ammonia, glucose, and creatine kinase after 15-min swimming exercise. Exercise training combined with AM supplementation increased endurance exercise capacity and increased hepatic and muscle glycogen content. AM reduced exercise-induced accumulation of the byproducts blood lactate and ammonia with acute exercise challenge. Moreover, we found no deleterious effects from AM treatment. Therefore, AM supplementation improved exercise performance and had anti-fatigue effects in mice. It may be an effective ergogenic aid in exercise training.
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Molecules 2014, 19, 2793-2807; doi:10.3390/molecules19032793
ISSN 1420-3049
Astragalus membranaceus Improves Exercise Performance and
Ameliorates Exercise-Induced Fatigue in Trained Mice
Tzu-Shao Yeh 1, Hsiao-Li Chuang 2,†, Wen-Ching Huang 3,†, Yi-Ming Chen 4,, Chi-Chang Huang 4,*
and Mei-Chich Hsu 5,*
1 School of Nutrition and Health Sciences, Taipei Medical University, Taipei 11031, Taiwan
2 National Laboratory Animal Center, National Applied Research Laboratories, Taipei 11529, Taiwan
3 Graduate Institute of Athletics and Coaching Science, National Taiwan Sport University,
Taoyuan 33301, Taiwan
4 Graduate Institute of Sports Science, National Taiwan Sport University, Taoyuan 33301, Taiwan
5 Department of Sports Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan
These authors contributed equally to this work.
* Authors to whom correspondence should be addressed; E-Mails: (C.-C.H.); (M.-C.H.); Tel.: +886-3-328-3201 (ext. 2619) (C.-C.H.);
+886-7-312-1101 (ext. 2793-613) (M.-C.H.).
Received: 15 February 2014; in revised form: 24 February 2014 / Accepted: 24 February 2014 /
Published: 3 March 2014
Abstract: Astragalus membranaceus (AM) is a popular “Qi-tonifying” herb with a long
history of use as a Traditional Chinese Medicine with multiple biological functions.
However, evidence for the effects of AM on exercise performance and physical fatigue is
limited. We evaluated the potential beneficial effects of AM on ergogenic and anti-fatigue
functions following physiological challenge. Male ICR strain mice were randomly
assigned to four groups (n = 10 per group) for treatment: (1) sedentary control and vehicle
treatment (vehicle control); (2) exercise training with vehicle treatment (exercise control);
and (3) exercise training with AM treatment at 0.615 g/kg/day (Ex-AM1) or
(4) 3.075 g/kg/day (Ex-AM5). Both the vehicle and AM were orally administered for 6 weeks.
Exercise performance and anti-fatigue function were evaluated by forelimb grip strength,
exhaustive swimming time, and levels of serum lactate, ammonia, glucose, and creatine
kinase after 15-min swimming exercise. Exercise training combined with AM
supplementation increased endurance exercise capacity and increased hepatic and muscle
glycogen content. AM reduced exercise-induced accumulation of the byproducts blood
Molecules 2014, 19 2794
lactate and ammonia with acute exercise challenge. Moreover, we found no deleterious
effects from AM treatment. Therefore, AM supplementation improved exercise
performance and had anti-fatigue effects in mice. It may be an effective ergogenic aid in
exercise training.
Keywords: Huangqi; exhaustion exercise; lactate; ammonia; glycogen
1. Introduction
Astragalus membranaceus (AM) is a well-known the “Qi-tonifying” or adaptogenic herb used in
Traditional Chinese Medicine. It has been prescribed for centuries for general debilitation and chronic
illnesses and to increase overall vitality [1]. The main constituents of AM roots are polysaccharides,
saponins, flavonoids, amino acids and trace elements [2,3]. More than 40 constituents in Astragalus
saponins have been identified from the Astragalus root [4]. Astragalosides have various
pharmacological activities and are used as a quality-control marker of AM [5]. In recent decades,
molecular features and pharmacokinetics and pharmacological actions of Astragaloside polysaccharides
and other active ingredients from the plant have been studied extensively, which could enable its
clinical use [6]. AM has effects against myocardial damage [7–10], promotes angiogenesis [11,12] and
protects endothelial function [13], prevents ultraviolet A-induced photoaging [14], is hepatoprotective [15],
improves insulin resistance [16] and is involved in immunomodulatory activities [17–19]. However,
few studies have directly addressed the possible anti-fatigue function of AM.
Fatigue is a complex phenomenon that can be defined as the inability to maintain expected muscle
strength, leading to reduced performance during prolonged exercise and has classified physical and/or
mental fatigue [20]. Physical fatigue is also called peripheral fatigue, which can be derived from the
action of the muscles and may be accompanied by deterioration in functional performance [21–23].
Chronic fatigue can lead to severe health problems [24,25]. There are at least two mechanisms that can
explain the occurrence of physical fatigue: oxidative stress and energy exhaustion [26]. Overloading
work or exhaustive exercise can lead to the accumulation of excess reactive free radicals, which result
in tissue damage. Exhaustion theory suggests that energy source depletion and excess metabolite
accumulation lead to fatigue [27]. However, several studies have shown that exogenous nutrition
supplement can reduce exercise-induced physical fatigue [28–32]. Research into specific nutrients or
herbal supplements is needed to find agents that reduce metabolite production and/or improve
energy utilization.
Herbal medicines and food factors have been investigated as an important resource for postponing
fatigue, accelerating the elimination of fatigue-related metabolites, and improving exercise
performance. AM has long been considered a potent remedy for regulating the body balance, with few
adverse effects, but scientific evidence of its action is lacking. AM may be an anti-fatigue herbal
supplement candidate. Here, we examined whether AM supplementation before intensive aerobic
exercise training could change body composition, physical activities, and physiologic features in vivo
in mice.
Molecules 2014, 19 2795
2. Results and Discussion
2.1. Body Weight and Other Metabolism-Related Organ Weights
Morphological data from each experimental group are listed in Table 1. Initial and final body
weights did not differ among treatment groups. The food intake was higher by 1.12-, 1.10 and
1.05-fold (p < 0.05), respectively, for the exercise control, Ex-AM1 and Ex-AM5 groups, than the
vehicle control group. The groups did not differ in weights of liver, kidney, epididymal fat pad,
muscle, and brown adipose tissue. The relative tissue weight (%) is a measure of different tissue
weights adjusted by individual body weight, and relative liver weight was lower for the Ex-AM5 group
than the vehicle group (p < 0.05).
Table 1. General characteristics of the experimental groups.
Characteristic Vehicle Control Exercise Control Ex-AM1 Ex-AM5
Initial body weight (g) 25.43 ± 0.24 25.47 ± 0.20 25.53 ± 0.29 25.49 ± 0.28
1 week body weight (g) 29.94 ± 0.57 29.11 ± 0.40 28.76 ± 0.50 28.78 ± 0.57
2 week body weight (g) 31.54 ± 0.59 30.07 ± 0.49 31.55 ± 0.50 31.41 ± 0.68
3 week body weight (g) 32.63 ± 0.48 32.43 ± 0.50 32.84 ± 0.48 33.26 ± 0.85
4 week body weight (g) 33.52 ± 0.44 33.53 ± 0.41 33.52 ± 0.53 34.23 ± 0.85
5 week body weight (g) 34.50 ± 0.45 33.68 ± 0.33 34.59 ± 0.56 35.11 ± 0.81
Final body weight (g) 35.59 ± 0.51 35.80 ± 0.46 36.28 ± 0.44 37.18 ± 0.76
Food intake (g/day) 6.78 ± 0.01 a 7.61 ± 0.01 d 7.44 ± 0.03 c 7.11 ± 0.03 b
Food efficiency (%) 1.49 ± 0.06 1.35 ± 0.05 1.44 ± 0.05 1.64 ± 0.08
Liver (g) 2.16 ± 0.03 2.14 ± 0.03 2.05 ± 0.03 2.06 ± 0.04
Kidney (g) 0.61 ± 0.01 0.65 ± 0.02 0.62 ± 0.02 0.63 ± 0.01
Epididymal fat pads (g) 0.52 ± 0.03 0.47 ± 0.02 0.42 ± 0.04 0.46 ± 0.02
Muscle (g) 0.36 ± 0.01 0.36 ± 0.01 0.37 ± 0.01 0.37 ± 0.01
Brown adipose tissue (g) 0.13 ± 0.01 0.15 ± 0.01 0.16 ± 0.01 0.16 ± 0.01
Relative liver weight (%) 6.07 ± 0.07 b 5.97 ± 0.07 b 5.67 ± 0.07b 5.54 ± 0.11 a
Relative kidney weight (%) 1.72 ± 0.03 1.81 ± 0.04 1.71 ± 0.04 1.71 ± 0.04
Relative epididymal fat pads weight (%) 1.45 ± 0.06 1.30 ± 0.07 1.17 ± 0.12 1.23 ± 0.07
Relative muscle weight (%) 1.01 ± 0.02 1.02 ± 0.02 1.02 ± 0.02 1.00 ± 0.03
Relative brown adipose tissue weight (%) 0.37 ± 0.01 0.42 ± 0.02 0.43 ± 0.03 0.43 ± 0.03
Data are mean ± SEM for n = 10 mice in each group. Data in the same row with different superscript letters
(a, b, c and d) differ significantly, p < 0.05, by one-way ANOVA. Food efficiency (%): body weight
gain (g/day) food intake (g/day) 100%. Muscle mass includes both gastrocnemius and soleus muscles in
the back part of the lower legs.
2.2. Effects of AM on Forelimb Grip Strength
The forelimb grip strength of mice increased with Ex-AM5 supplementation than with the vehicle
treatment (Figure 1A). On trend analysis, grip strength dose-dependently increased with AM dose
during training intervention (p < 0.005). Thus, high dose of AM may contribute to physiological activities.
Molecules 2014, 19 2796
Figure 1. Effect of A. membranaceus (AM) supplementation on forelimb grip strength (A)
and swimming exercise performance (B). Data are mean ± SEM of n = 10 mice in each
group by one-way ANOVA. * p < 0.05; *** p < 0.001.
2.3. Effect of AM on Exercise Performance in Weight-loaded Swim Test
Exercise endurance is an important variable in evaluating anti-fatigue treatment. Exercise endurance
in mice with a swim test increased with Ex-AM1 and Ex-AM5 supplementation than with the vehicle
treatment (Figure 1B). At higher AM doses, exercise performance was significantly longer, by
2.33-fold (p < 0.05), with Ex-AM5 compared to exercise control. On trend analysis, exercise
performance dose-dependently increased with AM dose (p < 0.005). Therefore, exercise training
combined with AM supplementation significantly increased exercise performance.
2.4. Effect of Exercise Training Combined with AM Supplementation on the Serum Levels of Lactate,
Ammonia, Glucose and Creatine Kinase (CK) After Acute Exercise Challenge
Muscle fatigue after exercise can be evaluated by biochemical indicators including lactate,
ammonia, glucose and CK levels after exercise [30]. Lactate levels decreased with Ex-AM5
supplementation than with the vehicle or exercise only treatment (Figure 2A). Serum ammonia levels
decreased with Ex-AM1 and Ex-AM5 supplementation than with the vehicle or exercise only
treatment (Figure 2B). Serum glucose contents increased with Ex-AM5 supplementation than with the
vehicle treatment (Figure 2C). Serum CK activity, a muscular damage marker, decreased with
Ex-AM5 supplementation among four groups (Figure 2D). Trend analysis revealed that AM treatment
had a significant dose-dependent effect on increasing blood glucose level (p < 0.001) and decreasing
serum levels of lactate and ammonia and CK (p < 0.001).
Molecules 2014, 19 2797
Figure 2. Effect of AM supplementation on serum lactate (A), ammonia (B), glucose (C),
and CK (D) levels after a 15-min swim test without weight loading. Data are mean ± SEM
of n = 10 mice in each group by one-way ANOVA. Different letters indicate a significant
difference p value. * p < 0.05; ** p < 0.01; *** p < 0.001.
2.5. Effect of AM Supplementation on Hepatic and Muscle Glycogen Levels
Hepatic and muscle glycogen levels increased with exercise control, Ex-AM1 and Ex-AM5
treatment (Figure 3). In addition, the trend analysis revealed that AM treatment had a significant
dose-dependent effect on increasing hepatic and muscle glycogen levels (p < 0.001).
2.6. Effect of AM Supplementation on Biochemical Analyses at the End of the Experiment
We examined whether AM treatment for 6 weeks could have negative effects on other biochemical
markers in healthy mice. We examined the liver- and kidney-related biochemical parameters and major
organs including liver, skeletal muscles, heart, kidney, lungs, and testes according to histopathological
examinations in AM-treated mice (Table 2 and Figure 4). We found no indication of a deleterious
effect with AM treatment. With exercise training and continuous AM supplementation for 6 weeks,
triglycerides level was significantly decreased by about 44% and 40% (p < 0.05) for the Ex-AM1 and
Ex-AM5 groups, respectively, as compared with the vehicle control. AM may enhance the effect of
exercise to reduce hyperlipidemia.
Molecules 2014, 19 2798
Figure 3. Effect of AM supplementation on levels of hepatic glycogen (A) and muscle
glycogen (B). Data are mean ± SEM of n = 10 mice in each group by one-way ANOVA.
*** p < 0.001.
Tab l e 2 . Biochemical analysis of the AM treatment groups at the end of the experiment.
Parameter Vehicle Control Exercise Control Ex-AM1 Ex-AM5
AST (U/L) 62.90 ± 3.17 68.90 ± 3.78 60.70 ± 2.93 59.00 ± 1.97
ALT (U/L) 42.10 ± 2.84 a 54.30 ± 2.34 b 39.20 ± 1.79 a 46.30 ± 1.93 a,b
ALP (U/L) 48.80 ± 3.22 63.40 ± 5.80 54.80 ± 3.59 59.20 ± 3.49
LDH (U/L) 301.10 ± 19.06 273.00 ± 23.09 254.70 ± 17.29 293.10 ± 15.41
Albumin (g/dL) 3.56 ± 0.08 3.76 ± 0.06 3.59 ± 0.05 3.73 ± 0.06
TBIL (μg/dL) 0.19 ± 0.03 0.22 ± 0.03 0.23 ± 0.03 0.22 ± 0.02
TP (g/dL) 4.73 ± 0.06 4.63 ± 0.06 4.56 ± 0.05 4.58 ± 0.05
BUN (mg/dL) 23.43 ± 0.77 23.07 ± 1.00 20.18 ± 0.57 23.21 ± 0.46
Creatinine (mg/dL) 0.13 ± 0.01 0.12 ± 0.01 0.11 ± 0.00 0.12 ± 0.01
UA (mg/dL) 1.43 ± 0.11 a 0.83 ± 0.05 b 1.33 ± 0.07 a 1.18 ± 0.09 a,b
TG (mg/dL) 228.00 ± 21.03 a 184.40 ± 20.34 a,b 126.70 ± 5.74 b 136.60 ± 10.02 b
TC (mg/dL) 110.60 ± 4.17 104.40 ± 4.45 112.80 ± 4.68 107.90 ± 4.13
Glucose (mg/dL) 179.80 ± 6.03 182.30 ± 6.73 181.00 ± 4.40 177.20 ± 4.56
Data are mean ± SEM for n = 10 mice in each group. Data in the same row with different superscript letters
(a and b) differ significantly, p < 0.05, by one-way ANOVA. AST, aspartate aminotransferase; ALT, alanine
aminotransferase; ALP, alkaline phosphatase; LDH, lactate dehydrogenase; TBIL, total bilirubin; TP, total
protein; BUN, blood urea nitrogen; UA, uric acid; TG, triacylglycerol; TC, total cholesterol.
2.7. Effect of AM Supplementation on Histological Examinations at the End of the Experiment
As shown in Figure 4, the four groups did not differ in histological observations of liver, muscle,
heart, kidney, lung, and testis.
Molecules 2014, 19 2799
Figure 4. Effect of AM supplementation on morphology of liver (A), skeletal muscle (B),
heart (C), kidney (D), lungs (E), and testes (F) tissues. Specimens were photographed
under a light microscope. (H&E stain, magnification: ×200, Scale bar, 40 μm).
2.8. Discussion
Previously, the effect of AM on exercise-induced accumulation of products of metabolism and
physical toxicity has been unclear. In this study, we compared the fatigue-alleviating effects of two
doses of AM as well as vehicle and exercise control on endurance in exercised and weight-loading
mice. We found that: (1) Exercise training combined with AM supplementation increased endurance
with exercise and increased hepatic and muscle glycogen content; (2) AM reduced exercise-induced
accumulation of byproducts such as blood lactate and ammonia by acute exercise challenge;
and (3) daily AM administration for 6 weeks had no toxic effects according to biochemical parameters
and histopathological examination. Thus, AM may have ergogenic and anti-fatigue functions.
Astragalus polysaccharides are the main active constituents of AM. The Astragalus polysaccharide
were found to have a positive effect on skeletal muscle for glucose homeostasis through stimulation of
protein kinase B (PKB)/glucose transporter 4 (GLUT4) pathways [33]. It has been demonstrated that
skeletal muscle 2-deoxyglucose uptake during muscle contractions is directly related to muscle GLUT-4
Molecules 2014, 19 2800
protein content [34] and GLUT-4-mediated muscle glucose transport does not limit exercise-stimulated
muscle glucose uptake [35]. One previous study reported that Astragalus polysaccharide increased the
number of GLUT4 transporters at the muscle cell surface [33]. In our study, exercise with AM
supplement significant increased exercise performance, and AM treatment had a significant dose-
dependent effect on increasing blood glucose after a 15-min swimming test without weight-loading.
These results indicate that AM may enhance muscle glucose uptake during exercise and continue AM
supplement could prolong the time of exercise.
Serum level of CK is an important clinical biomarker of muscle damage, muscular dystrophy,
severe muscle breakdown, myocardial infarction, autoimmune myositides and acute renal failure.
High-intensity exercise challenge could physically or chemically cause tissue damage and muscular
cell necrosis [36]. Furthermore, reactive oxygen species (ROSs), like lactate anion and protons, have
been suggested to be implicated in oxidative skeletal muscle fatigue. It is reported that ROS alter such
transport systems as potassium transport and thus contribute to the onset of fatigue [37]. Under
oxidative stress-induced cellular injuries, the cell membrane integrity can be damaged, and cytosolic
enzymes will leak out into the serum. Those enzymes, including lactate dehydrogenase (LDH), CK,
myoglobin, aspartate aminotransferase (AST), alanine aminotransferase (ALT), etc., can be parameters
indicating tissue injury under high-intensity exercise challenge [36]. Astragalosides is another
bioactive components of AM and a well antioxidant to protect exercise-induced oxidative stress [38,39].
In our study, serum CK activity was significantly lower by 40.7% with Ex-AM5 treatment than the
exercise control (p < 0.001). These results indicated that AM supplementation could stabilize
membranes and attenuate muscle damage during exercise.
Energy storage and supply is another important factor related to exercise performance. With energy
expenditure during exercise, physical fatigue is mainly caused by energy consumption and deficiency [40].
Catabolized fat and carbohydrates are considered the main sources of energy during exercise in
skeletal muscles, and glycogen is the predominant source of glycolysis for energy production.
Therefore, glycogen storage directly affects exercise ability [41]. In our study, liver glycogen content
was significantly higher, by 1.06- and 1.21-fold (all p < 0.001), with Ex-AM1 and Ex-AM5,
respectively, than exercise control. In addition, muscle glycogen content was significantly higher by
1.01- and 1.17-fold (all p < 0.001) with Ex-AM1 and Ex-AM5, respectively, than exercise control. The
glycogen-sparing effect of AM could provide an important survival advantage in situations requiring
extended periods of prolonged exercise endurance because glycogen depletion is associated with
physical exhaustion, and slower utilization of glycogen results in improved exercise endurance. As one
of the sources of blood glucose, liver glycogen plays an important role in controlling the availability of
cellular energy. It is possible that AM may have promoted glycogenolysis restraint and/or gluconeogenesis.
The levels of serum glucose, lactate, ammonia, and glutamine are known to serve as indicators of
accumulated fatigue and stress caused by exercise [42]. Blood lactate is the glycolysis product of
carbohydrate under anaerobic conditions, and glycolysis is the main energy source for short term
intensive exercise. Because the accumulation of blood lactate causes fatigue during physical exercise,
rapid removal of lactate is beneficial to relieving fatigue [43]. Ammonia, a metabolite of proteins and
amino acids, was linked to fatigue [44]. An increase in ammonia in response to exercise can be
managed by the use of glutamine and/or carbohydrates that interfere with ammonia metabolism [45].
The increase in ammonia level is related to both peripheral and central fatigue during exercise [23].
Molecules 2014, 19 2801
In our study, serum ammonia levels were significantly lower by 12.7% and 22.3% with Ex-AM1 and
Ex-AM5, respectively, than the exercise control (p < 0.001). Lactate levels were significantly lower by
25.6% (p < 0.05) with Ex-AM5 than exercise control. This result suggests that AM supplementation
may reduce exercise-induced byproducts accumulate and alleviate physical fatigue.
Together, our results suggest that AM supplementation may ameliorate exercise-induced oxidative
stress, stimulate blood circulation, improving the transport efficiency to nutritional minerals and
assisting excretion and the elimination of the by-products of metabolism, and thereby against
physical fatigue.
3. Experimental
3.1. Experiment Design
The AM used for supplementation in the study was purchased from Sun Ten Pharmaceutical Co.,
Biotechnology Ltd. (New Taipei, Taiwan). Male ICR strain mice (4 weeks old) grown under specific
pathogen free conditions were purchased from BioLASCO (Yi-Lan, Taiwan). One week of
acclimation to the environment and diet was allowed before the experiment began. All animals were
fed a standard laboratory diet (No. 5001; PMI Nutrition International, Brentwood, MO, USA) and
housed at room temperature (23 ± 1 °C), with 50%–60% humidity and lighting (lights on from 06:00
to 18:00). The Institutional Animal Care and Use Committee (IACUC) of National Taiwan Sport
University approved all animal experiments in this study, and the study conformed to the guidelines of
protocol IACUC-10206 approved by the IACUC ethics committee.
The recommended use of AM for humans is about 3 g per one intake with a normal diet. The mouse
AM dose (0.615 g/kg) used in this study was converted from a human equivalent dose (HED) based on
body surface area by the following formula from the US Food and Drug Administration: assuming a
human weight of 60 kg, the HED for 3 (g) 60 (kg) = 0.05 × 12.3 = a mouse dose of 0.615 g/kg; the
conversion coefficient 12.3 was used to account for differences in body surface area between a mouse
and a human as described in our recent study [46].
All animals were randomly assigned to four groups (n = 10 per group) for swim exercise training or
exercise with AM supplement treatment: (1) sedentary control and vehicle treatment (vehicle control);
(2) exercise training with vehicle treatment (exercise control); and (3) exercise training with 0.615 g/kg
AM (Ex-AM1) or (4) 3.075 g/kg AM (Ex-AM5). The vehicle group received the same volume of
solution equivalent to individual body weight. Both the vehicle and AM were given orally to each
animal for 6 weeks.
3.2. Swimming Exercise Training
Animals in the exercise control and Ex-AM1 and Ex-AM5 groups underwent an intensive aerobic
swim training program adapted from our recent study with some modifications [46]. They were placed
in a plastic container (65 cm high, 40 cm diameter) with 20-cm tap water depth maintained at 28 ± 1°C.
They trained 30 min on the first day, 45 min on the second day, then 60 min/day, 5 days/week. The
swim training was maintained for 1 h from weeks 2 to 6. After the first week, the swim training
consisted of 5 weekly sessions of 60 min of forced swimming with a 1% loading of body weight at
Molecules 2014, 19 2802
week 2. From weeks 3 to 4, animals underwent a 2% loading of body weight training protocol. At the
fifth and sixth week, the swimming load was up to 3% of body weight, which consisted of 5 weekly
swim sessions for 60 min each. Body weight was measured weekly, and the load was estimated and
increased accordingly.
3.3. Exhaustion Swimming Exercise Test
Swim to exhaustion exercise test involved constant loads corresponding to 5% of body weight to
evaluate endurance. The swimming exercise was carried out in a round tank (65 cm high, 40 cm
diameter), filled with water to 45 cm depth and maintained at a temperature of 28 ± 1 °C. To avoid
circadian variations in physical activity, swimming exercise was performed between 07:00 and 14:00,
when minimal variation in endurance capacity was confirmed in mice [47]. The endurance of each
mouse was recorded as the time from beginning swimming to exhaustion, which was determined by
observing loss of coordinated movements and failure to return to the surface within 7 s. Times floating,
struggling, and making necessary movements were considered in the swimming duration until
exhaustion and possible drowning.
3.4. Forelimb Grip Strength
A low-force testing system (Model-RX-5, Aikoh Engineering, Nagoya, Japan) was used to measure
forelimb grip strength of mice undergoing vehicle, exercise, and AM treatments. The amount of tensile
force was measured by use of a force transducer equipped with a mental bar (2 mm diameter and 7.5 cm
long) for each mouse in each group. As described in our previous studies [31,48,49], we grasped the
mouse by the base of the tail and lowered it vertically toward the bar. The mouse was pulled slightly
backwards by the tail while the two paws (forelimbs) grasped the bar, which triggered a “counter pull.”
This grip strength meter recorded the grasping force in grams. Forelimb grip strength testing was
performed after consecutive administration of the vehicle of AM for 6 weeks and 1 h after the last
treatment. The maximal force (in grams) recorded by the counter-pull of mice forelimbs was used as
grip strength.
3.5. Determination of Blood Biochemical Variables
The effects of AM on serum lactate, ammonia, and glucose levels, and CK activity were evaluated
post-exercise. At 1 h after the last administration, mice underwent a 15-min swimming test without
loading. After the swim exercise, blood samples were immediately collected from the submandibular
duct of pretreated mice and centrifuged at 1,500 ×g and 4 °C for 10 min for serum preparation.
Clinical biochemical assessment was determined by use of an autoanalyzer (Hitachi 7060, Hitachi,
Tokyo, Japan).
3.6. Tissue Glycogen Determination
Because liver and skeletal muscles are the 2 major tissues for glycogen deposition, we investigate
whether glycogen contents of these 2 target tissues could increase with AM administration. Mice
underwent treatment for 6 weeks and then were killed 1 h after the last treatment administration. The
Molecules 2014, 19 2803
muscle and liver were excised and weighed for a glycogen content analysis. The muscle and hepatic
glycogen levels were measured as described in our previous studies [31,48,49]. For each mouse, 60 mg
muscle and liver tissue was finely cut, weighed and homogenized in 0.3 mL cold 10% perchloric acid.
After centrifugation for 15 min with 15,000 ×g at 4 °C, the supernatant was carefully decanted and
kept on ice for analysis. A standard glycogen (Sigma, Linkou Dist., New Taipei City, Taiwan) or tissue
extract, 30 μL, was added to 96-well microplates, and iodine-potassium iodide reagent, 200 μL, was
added to each well for binding iodine to glycogen. An amber-brown compound developed immediately
after the reaction. Absorbance was measured at wavelength 460 nm with use of an ELISA reader
(Tecan Infinite M200, Tecan Austria, Austria) after the material rested for 10 min.
3.7. Histological Staining of Tissues
Fresh liver, skeletal muscles, heart, kidney, lungs, and testes tissues were collected and fixed in
10% formalin after mice were killed. Tissues were embedded in paraffin and cut into 4-μm thick slices
for morphological and pathological evaluation as we described previously [31,48,49]. Tissue sections
were stained with hematoxylin and eosin (H&E) and examined under a light microscope equipped with
a CCD camera (BX-51, Olympus, Tokyo, Japan) by a clinical pathologist.
3.8. Analysis of Astragalus Membranaceus by HPLC/CAD
To confirm the quality of AM, we analysed the main chemical constituents of AM. The quantitative
analysis of AM was confirmed using standard astragalosides I, II, III, and IV (ChromaDex, Irvine, CA,
USA) diluted with methanol and then sonicated for 30 min. The mixed standard was further diluted as
necessary to create a calibration curve. AM was extracted using methanol and sonicated for 30 min,
and was subsequently equilibrated at room temperature. An aliquot of each sample solution was then
filtered using a 0.45-μm PTFE syringe filter; the filtrate was collected in an HPLC vial for analysis.
The AM component was determined to contain 1.455 mg/g of total Astragaloside (Table 3).
Table 3. The Astragaloside compounds in A. membranaceus (AM).
A. membranaceus (mg/g)
Astragaloside I 1.02
Astragaloside II 0.24
Astragaloside III BRL
Astragaloside IV 0.195
Total Astragalosides 1.455
Astragalosides analysis by HPLC/CAD. Chromatographic condition: spectra were obtained by
scanning UV-Vis. A Phenomenex Kinetix C18 column (150 × 4.6 mm, 2.6 μm, 100 Å) was used. The
flow rate was set at 0.9 mL/min, injection volume was 5 μL, and temperature was set at 40 °C.
The CAD detection was used corona aerosol discharge detector. BRL, compound detected below
reporting limit.
Molecules 2014, 19 2804
3.9. Statistical Analysis
All data are expressed as the mean ± SEM. Statistical differences among groups were analyzed by
one-way ANOVA and the Cochran-Armitage test for dose-effect trend analysis with SPSS 14.0 (SPSS,
Chicago, IL, USA). In case of significant F ratios, Scheffe post-hoc tests were used to determine
differences. Statistical significance was set at p < 0.05.
4. Conclusions
A. membranaceus (AM) has anti-fatigue activity by decreasing serum lactate and ammonia levels
and increasing liver and muscle glycogen deposition, thereby promoting exercise performance in mice.
Although the detailed anti-fatigue mechanisms of AM remain to be elucidated, this study provides
science-based evidence to support traditional claims of anti-fatigue results with AM treatment and
suggests a use for AM as an ergogenic and anti-fatigue agent.
The study was funded by the National Science Council, Taiwan (NSC-99-2410-H-179-006-MY2
and NSC101-2410-H-037-016-MY3). The authors are grateful to Chin-Shan Ho for technical assistance
in measuring forelimb grip strength. We also thank Laura Smales for carefully reading the manuscript.
Author Contributions
Conceived and designed the experiments: Chi-Chang Huang and Mei-Chich Hsu. Performed the
experiments: Tzu-Shao Yeh, Wen-Ching Huang and Yi-Ming Chen. Analyzed the data: Tzu-Shao Yeh
and Hsiao-Li Chuang. Contributed reagents/materials/analysis tools: Hsiao-Li Chuang, Chi-Chang
Huang and Mei-Chich Hsu. Wrote and Revised the manuscript: Chi-Chang Huang and Mei-Chich Hsu.
Conflicts of Interest
The authors declare no conflict of interest.
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... Astragalus membranaceus Bunge, a commonly used traditional medicine and an immunomodulatory agent for fatigue, diarrhea, anorexia, and viral infections in China and Republic of Korea [31][32][33], has significantly superior Th1 regulatory effects compared to other natural products [33]. A. membranaceus extract activates IFN-γ and decreases IL-4 content in pediatric asthma patients [34] and significantly inhibits the increase in Th2responsive cytokine content in asthma experimental animal models [35]. ...
... It also decreases IL-4 content in forced swimming exercise and diet-restricted rat models [36]. Although A. membranaceus is used for athletes' adaptability and anti-fatigue activity through immunomodulatory effects [31] using a CPA-induced immunosuppressed mouse model, a commonly used animal-model for immunomodulation and anti-mutagenicity evaluation [11,13,16,29,30], its anti-mutagenic activity, which is a problem in chemotherapeutic agents including CPA, is rarely reported. Therefore, it is necessary to evaluate the systematic immunomodulatory effects, including anti-mutagenicity. ...
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Although the immunomodulatory effects of Astragali Radix extract (AR) have been documented, its anti-mutagenic activity, a problem arising from chemotherapeutic agents, is rarely reported. Therefore, the anti-mutagenic and immunomodulatory effects of AR were investigated using a cyclophosphamide (CPA)-induced immunosuppressed mouse model to develop an alternative immunomodulatory agent. The fluid-bed-dried aqueous extract of AR containing 37.5% dextrin and exopolymers purified from Aureobasidium pullulans SM-2001 (EAP) were used in this study. The therapeutic potentials of AR at doses ranging from 100 mg/kg to 400 mg/kg was estimated by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) based cytotoxicity and splenocyte proliferation assay, body weight and lymphatic organ weight measurements, hematological measurements, serum and spleen cytokine level measurements, natural killer (NK) cell activity measurements, real-time RT-PCR expressions of splenic mRNA, a micronucleus test, histopathological observations, and immunohistochemical measurements. In CPA-treated mice, a clear immunosuppressive effect was observed for all tested parameters. However, the oral administration of AR (100, 200, and 400 mg/kg) showed dose-dependent and favorable inhibitory activities on CPA-induced immunosuppression and mutagenicity as compared to 200 mg/kg EAP. Furthermore, AR (100–400 mg/kg) up-regulated the nuclear factor kappa-light-chain-enhancer of activated B-cells (NF-κB) which are related to NK-, T-, and B-cell activation, with no critical cytotoxicity. The results of this study clearly demonstrate that AR at an appropriate oral dose could act as a potential alternative agent with significant anti-mutagenicity and immunomodulatory properties.
... Over the years, extensive research has been conducted on the chemical components of AR. It is known that AR root contains saponins, polysaccharides, flavonoids, amino acids, and trace elements [8]. Among these, saponins are the major active constituents, especially Astragaloside IV (AS-IV). ...
... Bge. (Yeh et al., 2014) Raising contractibility of skeletal muscle; increasing the activity of SOD and expression of a-action mRNA in skeletal muscle; inhibiting lipid peroxidation in blood and skeletal muscle. ...
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A third of the world’s population suffers from unexplained fatigue, hugely impacting work learning, efficiency, and health. The fatigue development may be a concomitant state of a disease or the side effect of a drug, or muscle fatigue induced by intense exercise. However, there are no authoritative guides or clinical medication recommendations for various fatigue classifications. Traditional Chinese medicines (TCM) are used as dietary supplements or healthcare products with specific anti-fatigue effects. Thus, TCM may be a potential treatment for fatigue. In this review, we outline the pathogenesis of fatigue, awareness of fatigue in Chinese and western medicine, pharmacodynamics mechanism, and substances. Additionally, we offer a comprehensive summary of fatigue and forecast the potential effect of novel herbal-based medicines against fatigue.
... [2] Astragalus membranaceus(AM), a well-known traditional Chinese medicine, has been used to improve muscle wasting-related disorders for a long history. AM can reduce fatigue and enhance endurance to improve athletic performance, [11,12] and its components like formononetin and calycosin benefit muscle atrophy improvement and cardiovascular protection. [13,14] Network pharmacology is a method that combines laboratory Medicine and clinical investigations with data handling to study and clarify the mechanisms of drug actions. ...
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Multiple system atrophy (MSA) is a fatal neurodegenerative disease, it causes functional degradation of multiple organs and systems throughout the body. Astragalus membranaceus (AM), a well-known traditional Chinese medicine, has been used to improve muscle wasting-related disorders for a long history. In this study, we used network pharmacology and molecular docking to predict the mechanism underlying AM for the treatment of MSA. We screened the active compounds of AM and its related targets, as well as the target proteins of MSA. We made a Venn diagram to obtain the intersecting targets and then constructed a protein-protein interaction network to find the core targets and build an active ingredient-target network map. After subjecting the intersecting targets to gene ontology and Kyoto encyclopedia of genes and genomes analysis, the binding ability of core compounds and core target proteins were validated by molecular docking. A total of 20 eligible compounds and 274 intersecting targets were obtained. The core components of treatment are quercetin, kaempferol, and isorhamnetin, and the core targets are TP53, RELA, and TNF. The main biological processes are related to cellular responses and regulation. Molecular functions are mainly associated with apoptosis, inflammation, and tumorigenesis. Molecular docking results show good and standard binding abilities. This study illustrates that AM treats MSA through multiple targets and pathways, and provides a reference for subsequent research.
... Both Western allopathic medicine and traditional Chinese medicine have developed approaches to the management of EF. The traditional Chinese medicine approach offers interventions that include various prescriptions [7], acupuncture [8], and other methods to treat EF [9]. Among these, catgut embedding acupuncture was shown to improve EF in human athletes in Yong Zhen Chen's study. ...
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To improve the phenomenon of exercise-induced fatigue that often occurs during horse racing, we previously studied the improvement in exercise tolerance by acupoint catgut embedding preconditioning in an exercise-induced fatigue rat model. We found that acupoint catgut embedding pretreatment effectively improved animal exercise tolerance. Here, by combining transcriptomics and metabolomics, we aimed to explore the underlying mechanisms of this improvement. We used blood biochemical detection combined with ELISA to detect triglyceride (TG), total cholesterol (TC), lactate dehydrogenase (LDH), high-density lipoprotein (HDL), alanine transaminase (ALT), aspartate aminotransferase (AST), and glucose (GLU), arachidonic acid (AA), and free fatty acid (FFA) content and found that acupoint embedding can correct FFA, AA, TG, LDH, and AST in the blood. We used RT-qPCR to measure the expression of genes in tissue from the quadriceps femoris muscle. We found that solute carrier family 27 member 2 (Slc27a2), fatty acid binding protein 1 (Fabp1), apolipoprotein C3 (Apoc3), and lipoprotein lipase (Lpl) genes in the peroxisome proliferator-activated receptor (PPAR) signaling pathway were important. The regulation of lipid metabolism through the PPAR signaling pathway was important for improving the exercise endurance of rats in our exercise-induced fatigue model. Therefore, we conclude that acupoint catgut embedding can not only promote body fat decomposition and reduce lactic acid accumulation but also promote the repair of tissue damage and liver damage caused by exercise fatigue. Acupoint catgut embedding regulates the PPAR signaling pathway by upregulating Lpl expression and downregulating Slc27a2, Fabp1, and Apoc3 expression to further improve body fat metabolism.
... Astragalosides are biologically active substances from the plant Astragalus, that could protect against CCl4-induced acute liver injury [13], PQ-induced lung injury [14], or IS-induced kidney injury [15] in mice or rats by ameliorating oxidative stress. Our previous study revealed that Astragalus membranaceus is a nutritional activator that promotes myogenesis by stimulating Akt/mTOR signaling in skeletal muscle cell lines [16] and can stabilize the sarcolemma, as well as preventing exercise-induced fatigue in rodents [17]. Although the wide use of astragalus as an antioxidant and anti-inflammatory agent has been well demonstrated, no research has been conducted on its efficacy in preventing prolonged inflammation and muscle repair following eccentric exercise-induced muscle damage. ...
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Astragalosides have been shown to enhance endurance exercise capacity in vivo and promote muscular hypertrophy in vitro. However, it remains unknown whether astragalosides supplementation can alter inflammatory response and enhance muscle recovery after damage in humans. We therefore aimed to evaluate the effect of astragalosides supplementation on muscle’s intrinsic capacity to regenerate and repair itself after exercise-induced damage. Using a randomized double-blind placebo-controlled cross-over design, eleven male participants underwent 7 days of astragalosides supplementation (in total containing 4 mg of astragalosides per day) or a placebo control, following an eccentric exercise protocol. Serum blood samples and variables related to muscle function were collected prior to and immediately following the muscle damage protocol and also at 2 h, and 1, 2, 3, 5, and 7 days of the recovery period, to assess the pro-inflammatory cytokine response, the secretion of muscle regenerative factors, and muscular strength. Astragalosides supplementation reduced biomarkers of skeletal muscle damage (serum CK, LDH, and Mb), when compared to the placebo, at 1, 2, and 3 days following the muscle damage protocol. Astragalosides supplementation suppressed the secretion of IL-6 and TNF-α, whilst increasing the release of IGF-1 during the initial stages of muscle recovery. Furthermore, following astragaloside supplementation, muscular strength returned to baseline 2 days earlier than the placebo. Astragalosides supplementation shortens the duration of inflammation, enhances the regeneration process and restores muscle strength following eccentric exercise-induced injury.
Herbal drugs are manufactured from the leaves, roots, seeds, bark, fruit, stems, or flowers of various plants that have medical characteristics or are assumed to have medicinal benefits. The usage of herbal supplements by athletes has skyrocketed in the last decade. Athletes and non-athletes are increasingly using herbal medicines to boost endurance and strength performance. Herbal adaptogens are used to improve attention, increase endurance in scenarios where fatigue is present, reduce the number of stress-related diseases and impairments in the body, improve physical stamina, strength, and energy levels, improve sexual dysfunction, restore cognitive performance that has been affected by stress, and maintain cortisol.The research method involved a preliminary search on Google Search, PubMed, OVID Medline, Embase, ScienceDirect, Web of Science, and Google Scholar databases where keywords such as “Herbal adaptogens, Endurance, Athletes, Ashwagandha, Tulsi, Turmeric, Muscle strength” were used.In this article, we have reviewed the top 8 global market frontrunners of herbal adaptogens based on source, namely ashwagandha, astragalus, cordyceps, ginseng, holy basil, Rhodiola roseas, schisandra, and turmeric, and their effect on the improvement in the performance of athletes like increase in the muscle mass, endurance, and recovery of the athlete and their possible side effects.
Background: Cycloastragenol (CAG) is a sapogenin derived from the main bioactive constituents of Astragali Radix (AR). However, the current research on CAG metabolism in vivo and in vitro is still inadequate, and the metabolite cluster is incomplete due to incomplete analysis strategy. Objective: The objective of this study was to screen and identify the metabolic behavior of CAG in vivo and in vitro. Methods: A simple and rapid analysis strategy based on UHPLC-Q-Exactive Orbitrap mass spectrometry combined with data-mining processing technology was developed and used to screen and identify CAG metabolites in rat body fluids and tissues after oral administration. Results: As a result, a total of 82 metabolites were fully or partially characterized based on their accurate mass, characteristic fragment ions, retention times, corresponding Clog P values, and so on. Among the metabolites, 61 were not reported in previous reports. These metabolites (6 metabolites in vitro and 91 in vivo) were generated through reactions of hydroxylation, glucuronidation, sulfation, hydrogenation, hydroxylation, demethylation, disopropylation, dehydroxylation, ring cleavage, and carboxyl substitution and their composite reactions, and the hydroxylation might be the main metabolic reaction of CAG. In addition, the characteristic fragmentation pathways of CAG were summarized for the subsequent metabolite identification. Conclusion: The current study not only clarifies the metabolite cluster-based and metabolic regularity of CAG in vivo and in vitro, but also provides ideas for metabolism of other saponin compounds.
Astragalus membranaceus is a widely used herbal medicine in Asia. It has been recognized as possessing various biological properties, however, studies on the activity of the A. membranaceus polysaccharide (AMP), a major component of A. membranaceus, on human peripheral blood dendritic cells (PBDCs) have not been thoroughly investigated. In this study, we found that AMP induced changes in dendritic morphology and the upregulation of activation marker expression and inflammatory cytokine production in human blood monocyte-derived dendritic cells (MDDCs). The AMP promoted the activation of both blood dendritic cell antigen 1⁺ (BDCA1⁺) and BDCA3⁺ PBDCs. AMP-induced secretion of cytokines in the peripheral blood mononuclear cells (PBMCs) was mainly due to PBDCs. Finally, activated BDCA1⁺ and BDCA3⁺ PBDCs by AMP elicited proliferation and activation of autologous T cells, respectively. Hence, these data demonstrated that AMPs could activate dendritic and T cells in human blood, and may provide a new direction for the application of AMPs in the regulation of human immunity.
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The mechanisms that account for the strength loss after contraction-induced muscle injury remain controversial. We present data showing that (1) most of the early strength loss results from a failure of excitation-contraction coupling and (2) a slow loss of contractile protein in the days after injury prolongs the recovery time. Keywords: strength, damage, calcium, contractile protein, sarcoplasmic reticulum, plasmalemma
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To verify the beneficial effects of whey protein (WP) supplementation on health promotion and enhancing exercise performance in an aerobic-exercise training protocol. In total, 40 male ICR mice (4 weeks old) were divided into 4 groups (n=10 per group): sedentary control with vehicle (SC) or WP supplementation (4.1 g⋅kg, SC+WP), and exercise training with vehicle (ET) or WP supplementation (4.1 g⋅kg, ET+WP). Animals in the ET and ET+WP groups underwent swim endurance training for 6 weeks, 5 days per week. Exercise performance was evaluated by forelimb grip strength and exhaustive swim time as well as by changes in body composition and biochemical parameters at the end of the experiment. ET significantly decreased final body and muscle weight and levels of albumin, total protein, blood urea nitrogen, creatinine, total cholesterol, and triacylglycerol. ET significantly increased grip strength; relative weight (%) of liver, heart, brown adipose tissue (BAT) and levels of aspartate aminotransferase (AST), alanine aminotransferase, alkaline phosphatase, lactate dehydrogenase (LDH), creatine kinase (CK), and total bilirubin. WP supplementation significantly decreased final body, muscle, liver, BAT, and kidney weight, and relative weight (%) of muscle, liver, and BAT as well as levels of AST, LDH, CK, and uric acid. In addition, WP supplementation slightly increased endurance time and significantly increased grip strength and levels of albumin and total protein. WP supplementation improved exercise performance, body composition and biochemical assessments in mice and may be an effective ergogenic aid in aerobic exercise training.
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The disease burden of diabetes mellitus is increasing throughout the world. The need for more potent drugs to complement the present anti-diabetic drugs has become an imperative. Astragalus membranaceus, a key component of most Chinese herbal anti-diabetic formulas, has been an important prospect for lead anti-diabetic compounds. It has been progressively studied for its anti-diabetic properties. Ethnopharmacological studies have established its potential to alleviate diabetes mellitus. Recent studies have sought to relate its chemical constituents to types 1 and 2 diabetes mellitus. Its total polysaccharides, saponins, and flavonoids fractions and several isolated compounds have been the most studied. The total polysaccharides fraction demonstrated activity to both types 1 and 2 diabetes mellitus. This paper discusses the anti-diabetic effects and pharmacological action of the chemical constituents in relation to types 1 and 2 diabetes mellitus.
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Astragaloside IV (AS-IV), one of the major active constituents of Astragalus membranaceus in Traditional Chinese Medicine, has been widely used to treat ischemic diseases. However, the potential mechanism is this action is unclear. In this study, we tested the hypothesis that AS-IV might promote angiogenesis through multiple signaling pathways. Our data indicate that AS-IV treatment promotes umbilical vein endothelial cells (HUVEC) proliferation, migration, and tube formation. AS-IV treatment also activates JAK2/STAT3 and ERK1/2 signaling pathways, and up-regulates endothelial nitric oxide synthase (eNOS) expression and nitric oxide (NO) production. AS-IV-induced angiogenesis in HUVECs is significantly blocked by specific kinase inhibitors. Our study indicated that AS-IV is a key regulator of NO and angiogenesis through the JAK2/STAT3 and ERK1/2 pathways, which provides a mechanistic basis for the potential use of this compound in the treatment of clinical ischemic diseases.
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To review the pharmacological effects and mechanisms of action of Astragaloside IV in Huangqi (Radix Astragali Mongolici). Aticles focusing on Astragaloside IV in English and Chinese in databases were collected and reviewed in order to summarize the latest extraction separation, pharmacokinetics, and the pharmacological effects of astrageloside IV. Protective effects of Astrageloside IV on the cardiovascular system, immune, digestive, nervous system were identified, and the action mechanisms were associated with regulation of the calcium balance, anti-oxydant, antiapoptosis, antivirus, and so on. Astrageloside IV has broad application prospects, especially in cardiovascular diseases, digestive diseases, cancer and other modern high incidence, high-risk diseases, and could be developed as a medicine.
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Resveratrol (RES) is a well-known phytocompound and food component which has antioxidative and multifunctional bioactivities. However, there is limited evidence for the effects of RES on physical fatigue and exercise performance. The purpose of this study was to evaluate the potential beneficial effects of trans-RES on fatigue and ergogenic functions following physiological challenge. Male ICR mice from four groups (n = 8 per group) were orally administered RES for 21 days at 0, 25, 50, and 125 mg/kg/day, which were respectively designated the vehicle, RES-25, RES-50, and RES-125 groups. The anti-fatigue activity and exercise performance were evaluated using forelimb grip strength, exhaustive swimming time, and levels of serum lactate, ammonia, glucose, and creatine kinase (CK) after a 15-min swimming exercise. The exhaustive swimming time of the RES-25 group (24.72 ± 7.35 min) was significantly (p = 0.0179) longer than that of vehicle group (10.83 ± 1.15 min). A trend analysis revealed that RES treatments increased the grip strength. RES supplementation also produced dose-dependent decreases in serum lactate and ammonia levels and CK activity and also an increase in glucose levels in dose-dependent manners after the 15-min swimming test. The mechanism was related to the increased energy utilization (as blood glucose), and decreased serum levels of lactate, ammonia, and CK. Therefore, RES could be a potential agent with an anti-fatigue pharmacological effect.
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A new cycloartane-type triterpene glycoside, agroastragaloside V (1) was isolated from the roots of Astragalus membranaceus. The structure was identified as 3-O-β-(2'-O-acetyl)-D-xylopyranosyl-6-O-β-D-glucopyranosyl-(24S)-3β,6α,24α,25-tetrahydroxy- 9,19-cyclolanostane, by means of spectroscopic methods, including HR-FAB/MS, 1D NMR (1H, 13C, DEPT), 2D NMR (gCOSY, gHSQC, gHMBC, NOESY), and IR spectroscopy. Four known cycloartane glycosides, namely, agroastragaloside I (2), agroastragaloside II (3), isoastragaloside II (4) and astragaloside IV (5) were also isolated. All isolated compounds were tested for the ability to inhibit LPS-induced nitric oxide production in RAW264.7 macrophages.
Ethnopharmacological relevance: Astragaloside IV (As IV) is one of the main effective components isolated from the traditional Chinese medical herb Astragalus membranaceus. The protective effect of Astragalus membranaceus on myocardial hypertrophy has been extensively proved. To test the hypothesis that Astragaloside IV can ameliorate the myocardial hypertrophy and inflammatory effect induced by β-adrenergic hyperactivity, we carried out in vivo and in vitro experiments. Material and methods: In in vivo study, the isoproterenol (Iso) (5 mg kg(-1) d(-1)) was used as a model of myocardial hypertrophy by intraperitoneal injection. SD rats were randomly assigned to following six groups: A: the control; B: Iso group; C: Iso plus As IV 20 mg kg(-1) d(-1); D: Iso plus As IV 40 mg kg(-1) d(-1); E: Iso plus As IV 80 mg kg(-1) d(-1); F: Iso plus Propranolol 40 mg kg(-1) d(-1). In in vitro study, cultured neonatal rat cardiomyocytes were pretreated with As IV (3, 10, 30 μ mol L(-1)), Propranolol (2 μ mol L(-1)) and BAY11-7082 (5 μ mol L(-1)) for 30 min, and then incubated with Iso (10 μ mol L(-1)) for 48 h. For the rats in each group, the heart mass index (HMI) and the left ventricular mass index (LVMI) were measured. To measure the transverse diameter of left ventricular myocardial cells (TDM), the hematoxylin-eosin (HE) staining method was applied. In addition, the volume and the total protein content of cardiomyocytes were measured, the mRNA expression of ANP and TLR4 were quantified by RT-PCR, the protein expression of TLR4, IκBα and p65 were quantified by Western blot, and the level of TNF-α and IL-6 were measured by ELISA. Results: In vivo: Comparing the Iso group to the control, the HMI, LVMI, TDM were significantly increased; the protein expression of TLR4 and p65 were increased, while the IκBα were decreased; the expression of ANP, TLR4 mRNA, and TNF-α, IL-6 in serum were significantly increased. These changes could be partly prevented by As IV and Pro. In vitro: the over-expression of the cell size, total protein content could remarkably down-regulated by As IV and Pro, and the results of RT-PCR, Western blot and ELISA were similar to those of in vivo. Conclusions: The results of these studies indicate that Astragaloside IV has good protective effect on myocardial hypertrophy induced by isoproterenol. More specifically, the cardioprotection is related to inhibiting the TLR4/NF-кB signaling pathway and the attenuating inflammatory effect.
Astragaloside IV (AS-IV) is one of the main active constituents of Astragalus membranaceus, which has various actions on the cardiovascular system. However, its electrophysiological mechanisms are not clear. In the present study, we investigated the effects of AS-IV on action potentials and membrane currents using the whole-cell patch clamp technique in isolated guinea-pig ventricular myocytes. AS-IV prolonged the action potential duration (APD) at all three tested concentrations. The peak effect was achieved with 1×10(-6) m, at which concentration AS-IV significantly prolonged the APD at 95% repolarization from 313.1±38.9 to 785.3±83.7 ms. AS-IV at 1×10(-6) m also enhanced the inward rectifier K(+) currents (IK1) and inhibited the delayed rectifier K(+) currents (IK). AS-IV (1×10(-6) m) strongly depressed the peak of voltage-dependent Ca(2+) channel current (ICaL) from -607.3±37.5 to -321.1±38.3 pA. However, AS-IV was not found to affect the Na(+) currents. Taken together, AS-IV prolonged APD of guinea-pig ventricular myocytes, which might be explained by its inhibition of IK. AS-IV also influences Ca(2+) signaling through suppressing ICaL.