Issue |
Wuhan Univ. J. Nat. Sci.
Volume 29, Number 4, August 2024
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Page(s) | 374 - 382 | |
DOI | https://doi.org/10.1051/wujns/2024294374 | |
Published online | 04 September 2024 |
Biology
CLC number: Q189
Anti-Acute Fatigue Effects of Ethanol Extract of Bidens pilosa L. and the Profiling of Antioxidant Index in ICR Mice
鬼针草(Bidens pilosa L.)醇提物在ICR小鼠上的抗急性运动疲劳效应及相关抗氧化指标变化
1
School of Physical Education, Yunnan Normal University, Kunming 650500, Yunnan, China
2
School of Life Sciences, Yunnan Normal University, Kunming 650500, Yunnan, China
3
School of Ecology and Environmental Science, Yunnan University, Kunming 650050, Yunnan, China
† Corresponding author. E-mail: gw_wang@163.com (WANG Gongwu); juncao@vip.163.com (CAO Jun)
Received:
24
February
2024
The effects of ethanol extract of Bidens pilosa L. (EEB) on acute exercise fatigue and its underlying biochemical mechanism were investigated in this study. Sixty adult male ICR mice were divided into control, model, vitamin C (VC) 100, EEB40, EEB80, and EEB160 groups, receiving VC (100 mg/kg) or EEB (40, 80, 160 mg/kg) for 28 days (intragastrically, I.G.). The mice underwent tail-suspension, elevated plus maze (EPM), rotarod, and loaded swimming tasks and biochemical indices were measured. There were no significant differences in body weight, tail suspension time, EPM open arm time/entries and serum cortisone levels among the groups. Compared with the model group, there was an increase in rotarod latency in the VC100/EEB80 groups and an increase in loaded swimming time in the EEB80/EEB160 groups. Furthermore, the haptic and muscle glycogen levels decreased in the model group, while the haptic glycogen levels increased in the all VC/EEB groups. Similarly, the serum lactate and creatinine levels increased in the model group, but decreased in lactate (except for EEB160), creatinine (except for EEB40) and lactate dehydrogenase in the EEB80 group. In the liver, malonaldehyde (MDA) and oxidized glutathione (GSSG) levels increased in the model group; however, glutathione reductase (GR) (except for EEB40), glutathione (GSH) and GSH/GSSG ratios increased, with GSSG levels decreasing in all VC/EEB groups. In the quadriceps, the GR levels increased in the model, whereas it decreased in the VC100, EEB40 and EEB80 groups. These results suggest that EEB has anti-acute fatigue effect, potentially attributed to mitigate metabolite accumulation, enhancing glycogen reserves, and fortifying the antioxidant mechanism.
摘要
本文探讨了鬼针草(Bidens pilosa L.)醇提物(EEB)的抗急性运动性疲劳作用及其生化机制。将60只成年雄性ICR小鼠分为对照组、模型组、维生素C(VC)100组、EEB40组、EEB80组和EEB160组, 分别给予VC(100 mg/kg)或EEB(40、80和160 mg/kg)灌胃28天, 检测动物在悬尾、高架十字迷宫(EPM)、转棒和力竭负重游泳任务中的行为学表现, 并检测其生化指标。结果表明, 小鼠体重、悬尾不动时间、EPM开臂时间和进入次数及血清皮质酮无组间差异; VC100、EEB80组转棒时间, EEB80、EEB160组力竭时间较模型组升高; 模型组肝糖原和肌糖原水平较对照组降低, 而VC/EEB组肝糖原水平较模型组升高; 同样, 模型组血清乳酸、肌酐水平较对照组升高, 而VC/EEB组乳酸(EEB160除外), 肌酐(EEB40除外), 和EEB80组乳酸脱氢酶水平较模型组降低。肝: 模型组丙二醛和氧化型谷胱甘肽(GSSG)水平较对照组升高, 而VC/EEB组(EEB40除外)谷胱甘肽还原酶(GR)水平及所有VC/EEB组还原型谷胱甘肽(GSH)、GSH/GSSG比值较模型组升高, 而GSSG水平较模型组降低。股四头肌: 模型组仅GR水平较对照组升高, VC/EEB组(EEB160除外)GR水平较模型组降低。结果提示, EEB具有抗急性疲劳作用, 可能与减少机体代谢产物的累积、增加糖原储备和增强抗氧化机制相关。
Key words: Bidens pilosa L. / anti-fatigue / loaded swimming / oxidative stress / liver glycogen / ICR mice
关键字 : 鬼针草 / 抗疲劳 / 力竭游泳 / 氧化应激 / 肝糖原 / ICR小鼠
Cite this article: WANG Xiaoqin, ZHANG Yaqin, WANG Gongwu, et al. Anti-Acute Fatigue Effects of Ethanol Extract of Bidens pilosa L. and the Profiling of Antioxidant Index in ICR Mice[J]. Wuhan Univ J of Nat Sci, 2024, 29(4): 374-382.
Biography: WANG Xiaoqin,female, Master, associate professor, research direction: sports physiology and neurobiology. E-mail: xiaoqinwang2001@163.com
Fundation item: Supported by the National Natural Science Foundation of China (31760278)
© Wuhan University 2024
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
0 Introduction
Bidens pilosa L., an annual herb belonging to the composite family, is widely distributed in tropical and temperate regions across the globe, boasting abundant resources[1]. Bidens pilosa L. comprises flavonoids, polyalkynes, terpenes, volatile oils, and other constituents, exhibiting pharmacological effects encompassing of anti-cancer, antioxidation, blood sugar reduction, blood pressure lowering, blood lipid lowering and cognitive improvement[2-5]. However, the potential anti-fatigue properties of the extract of Bidens pilosa L.remain unexplored. Exercise fatigue refers to the inability of the body's physiological function to maintain at a specific level and/or to maintain a predetermined intensity during exercise, and "exhaustion" exercise is a form of exercise fatigue[6]. Exhaustive exercise can rapidly consume the body's energy reserves, especially glycogen, resulting in strong oxidative stress reaction and even damage to cells and tissues[7]. Bidens pilosa L. is rich in flavonoids, alkynes and other components and has strong antioxidant effects[2,3,8,9]. Therefore, it is speculated that Bidens pilosa L. may protect cells through antioxidant mechanisms and play an anti-fatigue role. In this study, the effects of ethanol extract of Bidens pilosa L. (EEB) on acute exercise fatigue and profiling of oxidative stress-related biochemical index of exhausted mice were investigated, in order to promote the development of Bidens pilosa L. resources and provide animal experimental basis for anti-fatigue application.
1 Materials and Methods
1.1 Materials
1.1.1 Bidens pilosa L.
The Bidens pilosa L. was dry whole grass, which was collected from Chenggong, Kunming, Yunnan, China (24°N, 102°E) and identified by Botanist Li Zhimin. The specimens are preserved in the herbarium of the School of Life Sciences, Yunnan Normal University (YNNU). After drying and crushing, 65.01 g of the powder was extracted with 70% ethanol at a ratio of 1:20 for 60 min. The mixture was treated with ultrasound twice (60 ℃, 25 min/time), followed by filtration and evaporation under reduced pressure at 50 ℃ to obtain the EEB. The regression equation of the standard curve using rutin (HPLC (high performance liquid chromatography) grade ≥ 98%, standard substance from Shanghai Yuanye Bio-Technology Co. Ltd, Shanghai, China) as the standard product was Y = 22.672X + 0.009 4 (R2 = 0.999 7). The total flavonoid content of the EEB was 11.54 mg/mL and the raw material content was 1.2% (mass fraction) after averaging the results from two tests.
1.1.2 Animals
Sixty specific-pathogen-free male ICR mice, aged 4-5 weeks and weighing 25±2 g, were purchased from the Experimental Animal Center of Yunnan University (Kunming, China), license number: SCXK (Yunnan) K2021-0001. The mice were randomly divided into control group, model group, VC100 group (vitamin C, superior pure, Tianjin Kemiou Chemical Reagent Co., Ltd., Tianjin, China), EEB40 group, EEB80 group and EEB60 group. The EEB groups were respectively treated with 40, 80, and 160 mg/kg of EEB according to the corresponding dose; VC100 group was given VC at a dose of 100 mg/kg; control group and exhaustion group were given equal volume of solvent distilled water (DW), once a day (intragastrically, I.G.) for continuous 28 days. Before the experiment, the animals were acclimated in the laboratory for a week and allowed to breed freely. The lighting in the feeding room was 12 h/12 h alternating light and dark, and the temperature was maintained at 22±1 ℃. All operations involving experimental animals in this study were carried out in accordance with the Regulations on the Administration of Experimental Animals of the State Council of China and relevant management regulations. The experimental program has been approved by the Biomedical Ethics Committee of Yunnan Normal University. The experimental process is shown in Fig. 1.
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Fig. 1 The timeline of experiments |
1.2 Methods
1.2.1 Loaded swimming acute exercise fatigue model
On day 28 and day 38 of the experiment, exhaustive loaded swimming was conducted. A lead-tin alloy wire, equivalent to 5% of the mouse's body weight, was affixed to the base of its tail and immersed in a cylindrical acrylic water tank (height 31 cm, diameter 12 cm, water depth 23 cm) at a controlled water temperature of 25±1 ℃. The mice were considered submerged when their heads remained under water for a continuous duration of 5 s before being promptly retrieved. The time elapsed from entering the water to being pulled out was recorded. In contrast, mice in the control group swam without any additional weight for a fixed duration of only 60 s.
1.2.2 Rotarod test
On day 22, the animals underwent three sessions of adaptive rotarod training. During the first session, the rotational speed was gradually increased from 0 r/min to 20 r/min within 1 min and then maintained for 5 min. The second session took place after a 1-hour interval, where the rotational speed was escalated from 0 r/min to 25 r/min within 1 min and again sustained for another 5 min. Following another hour's rest, the third session was conducted with identical parameters as the second one. On the second day, a formal test was performed in which the rotational speed accelerated from 0 r/min to 25 r/min within 1 min while recording the latency for fatigue-induced falls in animals. A total of three tests were carried out with an intermediate resting period of 1 h. The control mice were solely placed on stationary rotating rods without engaging in any exercise.
1.2.3 Tail suspension test
On day 29, tail suspension test was performed. During the test, the mouse tail was attached to the fixed device with tape 1 cm away from the end of tail. There were partitions between the mice, and the distance between the mice and the surrounding partitions was ≥ 15 cm. The tail suspension was tested for 6 min, and the immobile time of the mice during the last 4 min was recorded.
1.2.4 Elevated plus maze (EPM) test
The EPM device utilized in this study comprises two open arms (30 cm long, 5 cm wide, and with an edge 1 cm high), two closed arms (30 cm long, 5 cm wide, and with a wall 15 cm high), as well as a square central area (5 cm on each side) positioned on a base plate located at a distance of 50 cm from the ground. To initiate the test, the mice were placed within the central area facing towards the open arms. An animal behavior analysis software (Xeye Maba3.2, Beijing MacroAmbition S&T Development Co. Ltd, Beijing, China) was employed to record the movement trajectory of each mouse within the maze for 5 min. The percentage of open arm entries (OE) and the open arm time (OT) were subsequently calculated using the recorded data. Following each mouse's trial, the mazes were cleaned using 75% ethanol.
1.2.5 Biochemical index determination
After the second load swim for 30 min, the mice were killed by deep anesthesia with 4% pentobarbital sodium (100 mg/kg, intraperitonelly, Merck, Darmstadt, Germany). The hearts were exposed, and blood was collected with a coagulant tube, centrifuged (4 ℃, 3 500 r/min, 5 min), and serum was taken for temporary storage at –80 ℃. Liver and gastrocnemius muscle tissues were taken on ice, homogenized and centrifuged rapidly according to kit requirements. The supernatant was taken and temporarily stored at –80 ℃. All samples were tested according to the instructions of the kit. Serum index: lactate (LA), lactate dehydrogenase (LDH), creatinine (CR) and blood urea nitrogen (BUN) were detected by micro/microplate methods. Serum cortisone was detected by Elisa method. Hepatic and gastrocnemius indices: hepatic glycogen (HG), muscle glycogen (MG), malonaldehyde (MDA), superoxide dismutase (SOD), glutathione reductase (GR), glutathione (GSH), and oxidized glutathione (GSSG) were all determined by microplate method (All kits were purchased from Shanghai Enzyme-linked Biotechnology Co., Ltd, China).
1.3 Data Statistical Analysis
All data of each group was shown as mean ± SEM (standard error of mean). The differences among groups were analyzed by one-way ANOVA combined with LSD (least significant difference) post hoc multiple comparison test. In case of non-normal distribution, Kruskal-Wallis non-parametric test combined with Dunn's post hoc test was applied for multiple comparisons. Paired sample t-test or repeated measures ANOVA were conducted to compare intra-group data before and after. A significance level of P < 0.05 denoted statistically significant differences.
2 Results
2.1 Effects of Exhaustive Exercise and EEB on Body Weight
The animals did not show serious depilation, anorexia, abnormal behavior or death. The body weight of mice in all groups increased significantly compared to the initial body weight (P < 0.01), but the difference among groups was not statistically significant (P > 0.05), indicating that the EEB had no effect on the increase of body weight of mice, as shown in Table 1.
Effect of EEB on the body weight of mice (unit:g)
2.2 Effect of EEB on Rotarod Latency and Loaded Swimming Time
Compared with the model group, the rotarod latency and exhaustive swimming time of mice in the VC100 and EEB groups improved. The rotarod duration of VC100 and EEB80 groups was significantly higher than that of model group (Fig. 2(a), all P < 0.05). In the loaded swimming test, the exhaustion time of EEB80 and EEB160 groups was significantly higher than that of model group (Fig. 2(b), all P < 0.01). The average exhaustion time of the three EEB groups increased by 13.65%, 60.3% and 130.95%, respectively (Fig. 2(c)).
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Fig. 2 Effects of EEB on the duration of rotarod test and loaded swimming in mice *P < 0.05, **P < 0.01 vs. model |
2.3 Effects of EEB on Emotion-Related Behaviors and Serum Cortisone Level
As shown in Table 2, there was no significant difference among groups in the immobility time of animals in the tail suspension task, the OT and OE in the EPM task, and the serum cortisone level (all P < 0.05).
Effects of EEB on performance of tail suspension, elevated plus-maze and serum cortisone level after loaded swimming
2.4 Effect of EEB on Glycogen Reserve (HG, MG)
As shown in Fig. 3, the HG (P < 0.05) and MG (P < 0.01) contents of the model group were significantly decreased compared with the control group, suggesting that the loaded swimming test can significantly consume carbohydrate energy reserves including HG and MG. Compared with the model group, HG contents in all VC100/EEB groups were significantly increased (Fig. 3(a), P < 0.05) but MG contents showed no significant change (Fig. 3(b), P > 0.05).
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Fig. 3 Effect of EEB on the hepatic glycogen and muscle glycogen after loaded swimming in mice *P < 0.05, **P < 0.01 vs. control; #P < 0.05, ##P < 0.01 vs. model |
2.5 Effects of EEB on the Profiling of Metabolism-Related Blood Biochemical Index in Loaded Swimming Mice
As shown in Fig. 4, compared with the control group, the blood CR level (Fig. 4(a), P < 0.01) was significantly increased, while the level of LA showed a slight increase (Fig. 4(c), P > 0.05). There were no differences in the BUN level and LDH activity among groups (Fig. 4(b) and Fig. 4(d), P > 0.05), except for LDH activity in the EEB80 group, which was lower than that in model group (P < 0.05). Compared with the model group, LA and CR levels in VC and EEB groups showed a decreasing trend, and LA levels in VC100, EEB40 and EEB80 groups, CR levels in VC100, EEB80 and EEB160 groups significantly decreased (Fig. 4(a), Fig. 4(c), all P < 0.05).
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Fig. 4 Effect of EEB on the profile of the serum biochemical index after loaded swimming in mice *P < 0.05, **P < 0.01 vs. control; #P < 0.05, ##P < 0.01 vs. model |
2.6 Effects of EEB on Profiling of Oxidative Stress Related Index (MDA, SOD, GR, GSH) in Mouse Liver or Quadriceps Femoris after Loaded Swimming
As shown in Fig. 5, the levels of hepatic MDA and GSSG in the model group exhibited significant increases compared to the control group (Fig. 5(a) and Fig. 5(e), all P < 0.01). Conversely, the levels of SOD (Fig. 5(b)), GR (Fig. 5(c)), and GSH (Fig. 5(d)) displayed a slight upward trend, consistent with a decrease in GSH/GSSG ratio (Fig. 5(f)). Compared to the model group, the GR (VC100, EEB80, and EEB160, all P < 0.05), GSH (all P < 0.01) and GSH/GSSG ratio (all P < 0.05) in all VC/EEB groups were significantly elevated. Additionally, GSSG showed a significant decrease in all VC/EEB groups (all P < 0.05 vs. model).
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Fig. 5 Effect of EEB on the profiling of oxidative stress-related biochemical index in mouse liver and quadriceps femoris after loaded swimming *P < 0.05, **P < 0.01 vs. control; #P < 0.05, ##P < 0.01 vs. model; &P < 0.05 vs. VC100 |
As shown in Fig. 5, the GR level of quadriceps femoris in the model group was significantly higher than that in the control group (P < 0.01). There were no significant differences in MDA, SOD, GSH and GSH/GSSG among groups (all P > 0.05). Compared with the model group, GR levels were significantly decreased in the EEB40 and EEB 80 groups (Fig. 5(c), all P < 0.01).
3 Discussion
3.1 EEB Treatment did not Interfere with Normal Growth of Mice
Results showed that after 28 d of EEB/VC treatment, the body mass of animals in all groups increased steadily, with no significant difference among groups, suggesting that the growth and development of mice were not affected (see Table 1). Frida et al[10] pointed out that the acute toxicity (LD50) of oral EEB to mice was 6.15 g/kg. The LD50 of mice with oral VC was 3 367 mg/kg. According to the classification standard "GB 15193.3-2014 Acute Oral Toxicity Test of National Food Safety Standard", EEB and VC can be classified as low toxicity or actual non-toxic substances. It can be seen that the maximum doses of EEB and VC used in this study (160 and 100 mg/kg) are far lower than their LD50 doses, indicating their general safety.
3.2 EEB Demonstrated Anti-Acute Exercise Fatigue Effects Without Emotion Disturbance
Anxiety and depression are two important emotional states. EPM[11] and tail suspension[11] are recognized as common behavioral tasks for detecting anxiety and depression-like behavior in rodents, respectively, and for screening anti-anxiety/depressive drugs. Cortisone is a major stress-related hormone in rodents. In this study, there were no differences between groups in suspended tail immobility time, the time and entry frequency of open arm in the EPM, and the serum cortisone level. Wang[12] demonstrated that serum cortisone levels in rats subjected to swimming load for 15 min and 1 h after exhaustion were significantly higher than that of immediately after exhaustion, suggesting a fluctuation in cortisol release with an initial decrease followed by an increase. In this study, cortisone was sampled 30 min after exhaustion, possibly reflecting a trough phase where mouse hormone levels had not significantly risen. These findings suggest that treatments with VC and EEB, as well as acute increases in physical activity such as exhaustive swimming and rotarod tests, may not induce significant changes in depressive or anxiety-like behavior in animals.
The rotarod task is a commonly used behavioral task to assess sensorimotor skill learning and endurance in animals[13]. Exhaustion loaded swimming test is a commonly used method for modeling or testing exercise fatigue[14]. In this study, animals in the VC100/EEB group showed significantly longer rotarod latency and loaded swimming time compared to those in the model group, suggesting that EEB, similar to conventional antioxidant VC, has certain anti-fatigue effects and can significantly improve the exercise endurance of animals (Fig. 2).
3.3 EEB Mainly Improved Hepatic Glycogen Reserve
Glycogen, mainly including HG and MG, serves as an important carbohydrate reserve involved in the body's energy supply. Therefore, a key indicator of exercise fatigue is the severe consumption of energy reserves such as HG and MG[15,16]. Previous studies have shown that ethanol extract of Moringa oleifera leaves[14], apple pomace polysaccharide[15], lemon seed flavonoids[17], Pinus koraiensis leaf extract[18], and fermented soybean protein peptide[19] can enhance glycogen levels and exhibit anti-fatigue effects. The low glycogen level of the model group (Fig. 3) suggests that exhaustive loaded swimming could reduce reserves of both HG and MG, resulting in acute exercise fatigue, which is consistent with previous reports[14-19]. In this study, only the VC and EEB groups were shown higher HG levels than the model group, suggesting that VC and EEB may exert anti-fatigue effects by reducing HG consumption rather than rapidly replenishing it. This finding is not completely consistent with the previous reports. The reason may be that samples were collected 30 min after exhaustive swimming in our study, while samples were collected after 9-day loaded swimming training in previous studies, for example, Bian et al[14], Lee et al[18] and Fang et al[19] conducted freely swimming on the second day after exhaustion test and then collected sample for 1 h after rest. Animals in these researches may have shown improved MG reserves due to adaptive training or longer recovery times. Overall, this study demonstrates that EEB has a protective effect on HG reserve, potentially contributing to its anti-fatigue mechanisms.
3.4 Effects of EEB on the Profiling of Metabolism-Related Blood Index
LA is a product of anaerobic glycolysis, and one of the key indicators of exercise fatigue[16,20,21]. The present results (Fig. 4) showed that the serum LA level was higher in the model group, while the serum LA content of VC100, EEB40 and EEB80 groups was significantly lower than that in the model group, indicating that 100 mg/kg of VC and 40 and 80 mg/kg of EEB could promote the clearance of increased LA level caused by acute fatigue. According to the "Lactate Shunt Theory", LA, as an important energy substance and signaling molecule, plays a role in information exchange and energy redistribution across cells and throughout the body[22]. In this study, both HG and MG were depleted with exhaustive exercise. EEB/VC treatment improved HG levels and rotarod/loaded swimming ability, possibly because EEB/VC promoted the full oxidation of LA in skeletal muscle, thereby enhancing the exercise ability. On the other hand, it may encourage blood circulation to transport LA to the liver, where it is converted back into glycogen via the gluconeogenic pathway for storage. This could explain the observed decrease in LA levels and increase in HG levels in the EEB/VC group. Based on the principle of conservation of energy, LA mainly comes from the anaerobic glycolysis pathway of MG metabolism. Thus, it can be inferred that the production of LA in EEB/VC group is not reduced, but more LA produced during exercise is fully utilized.
CR is the metabolic end product of creatine, an important energy substance in muscle. The level of blood CR is related to muscle fatigue and renal excretion dysfunction[23,24]. LDH is the final enzyme in the glycolysis pathway of muscle cells to produce LA. The level of LDH in the blood correlates with the leakage of LDH into the blood due to cell damage. In this study, acute exhaustive exercise significantly increased the serum CR level of mice (equivalent to 336% in the control group), while VC and EEB treatment significantly reduced the blood CR level. Additionally, a decrease in blood LDH activity was observed in the EEB80 group (Fig. 4). These results suggest that VC and EEB may play an anti-fatigue role by protecting cell integrity and thereby reducing serum CR and LDH.
3.5 Effects of EEB on the Profiling of Antioxidant Biochemical Index in Liver and Skeletal Muscle
MDA is the product of lipid peroxidation, which can reflect the degree of lipid peroxidation and cell damage. GSSG is the oxidized form of GSH, which can be converted between the two under the catalysis of GR. SOD is an important antioxidant metalloenzyme. In this study (Fig. 5), exhaustive exercise led to an increase in MDA and GSSG content in mouse liver tissue, which may indicate increased hepatic oxidative stress caused by acute fatigue. SOD, GSH and GR, as important antioxidant factors, showed an increasing trend in the model group, which could be regarded as a compensatory protective response by the body. VC and EEB enhanced liver GSH and GR levels and reduced GSSG levels, thus playing an antioxidant role in liver protection. Previous studies have shown that Bidens pilosa L. and its extracts can reduce MDA levels in liver tissue or serum and restore antioxidant enzymes such as SOD in liver injury/fibrosis or non-alcoholic fatty liver models of rodents, thereby playing a role in liver protection[25,26]. Our study proves that EEB can protect liver injury caused by acute exhaustive exercise by enhancing antioxidant capacity (SOD, GR) and reducing peroxide products (MDA, GSSG), thereby playing an anti-fatigue role.
As seen in Fig. 5, exhaustive exercise in this study mainly promoted an increase in skeletal muscle GR activity, while EEB (40 and 80 mg/kg) significantly decreased the GR level. However, this effect appeared to have little effect on GSH level and GSH/GSSG ratio. The level of skeletal muscle GR in the model group was significantly higher than that in the control group (136%). Previous reports suggest that both the human and mice of high-intensity/exhaustive exercise will show increased activity of antioxidant enzymes such as GR, which can compensate for the oxidative stress caused by exercise fatigue[7,27,28]. However, GR activity in skeletal muscle was inhibited after VC/EEB treatment, which may be due to the strong antioxidant effects of VC and EEB and the negative feedback regulation caused by the high reduced GSH content in muscle. On the other hand, the total amount of skeletal muscle is large, and the degree of participation in exhaustive exercise is heterogeneous in time and space, muscle fiber type and plasticity[29,30]. This study only tested the quadriceps sample, so the role of skeletal muscle in anti-fatigue of EEB needs more comprehensive investigation. These results suggest that EEB and VC may exert antioxidant and anti-fatigue effects by increasing the levels of liver antioxidant factors, inhibiting peroxide products, protecting the normal function of liver cells, and increasing HG content[7,27,28].
4 Conclusion
This study firstly proves that EEB has an anti-acute exercise fatigue effect in mouse. This effect may be related to the promotion of metabolite clearance (especially peroxides), improvement of antioxidant capacity for cell protection, and enhancement of glycogen reserves. Bidens pilosa L. shows potential application as an anti-fatigue drug or functional food, but its underlying mechanisms require further investigation.
Acknowledgments
We are grateful to Master candidates SHI Jingquan, XIE Huiheng & LI Yanrong, Ph.D. candidates GAO Runchi, TANG Xianghua & ZHAO Sanjun from YNNU for their assistance in animal feeding and equipment support.
Conflicts of Interest
The authors declare no conflicts of interest.
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All Tables
Effects of EEB on performance of tail suspension, elevated plus-maze and serum cortisone level after loaded swimming
All Figures
![]() |
Fig. 1 The timeline of experiments |
In the text |
![]() |
Fig. 2 Effects of EEB on the duration of rotarod test and loaded swimming in mice *P < 0.05, **P < 0.01 vs. model |
In the text |
![]() |
Fig. 3 Effect of EEB on the hepatic glycogen and muscle glycogen after loaded swimming in mice *P < 0.05, **P < 0.01 vs. control; #P < 0.05, ##P < 0.01 vs. model |
In the text |
![]() |
Fig. 4 Effect of EEB on the profile of the serum biochemical index after loaded swimming in mice *P < 0.05, **P < 0.01 vs. control; #P < 0.05, ##P < 0.01 vs. model |
In the text |
![]() |
Fig. 5 Effect of EEB on the profiling of oxidative stress-related biochemical index in mouse liver and quadriceps femoris after loaded swimming *P < 0.05, **P < 0.01 vs. control; #P < 0.05, ##P < 0.01 vs. model; &P < 0.05 vs. VC100 |
In the text |
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