Anti-fatigue effect of aqueous extract of Hechong (Tylorrhynchus heterochaetus) via AMPK linked pathway

doi:10.1016/j.fct.2019.111043

Food and Chemical Toxicology

Volume 135, January 2020, 111043

https://doi.org/10.1016/j.fct.2019.111043 Get rights and content

Highlights

•
Hechong presented a significant anti-fatigue effect via swimming tests.
•
Hechong increased GLU, TG and LDH levels and decreased LA, CK and BUN levels.
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Hechong improved the expressions of AMPK and PGC-1α in the liver and skeletal muscles.
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Hechong possesses an anti-fatigue effect through the AMPK-linked pathway.

Abstract

Tylorrhynchus heterochaetus (Hechong in Chinese) has been used in Chinese traditional medicine for treating various diseases. This study was aimed to assess the anti-fatigue effect of T. heterochaetus on Kunming mice and its primary mechanism of action using forced running, rotating rod and weight-loaded swimming tests. Low (2.70 mg/0.5 mL/20 g), medium (5.41 mg/0.5 mL/20 g) and high (6.58 mg/0.5 mL/20 g) doses of T. heterochaetus aqueous extract were treated to mice for 28 days. Among the doses, the low and medium doses showed significant (p ≤ 0.05) anti-fatigue effect on the weight-loaded swimming test. Also, T. heterochaetus extract showed significant (p ≤ 0.05) effects on fatigue-related blood parameters by increasing the GLU, TG and LDH levels and decreasing the LA, CK and BUN levels. The levels of liver and skeletal muscle glycogen were also significantly (p ≤ 0.05) increased after treatment. Further, on Western blot analysis, it has been found that T. heterochaetus enhanced the expressions of AMPK and PGC-1α in the liver and skeletal muscles of mice. From the study, our outcomes suggest that T. heterochaetus possess an anti-fatigue effect through the AMPK-linked pathway and thereby it can regularize the energy metabolism.

Graphical abstract

Introduction

Fatigue is a feeling of extreme tiredness, with decrease performance and physical and mental unfitness such as drowsiness, inattention, and distraction (Xu et al., 2017). Central and peripheral fatigue are the two types, which affects more than 20% of people worldwide (Zhao et al., 2017). Central fatigue is caused due to the factors related to the central nervous system, brain spinal cord and motor neurons whereas peripheral fatigue are related to the muscle strength weakness triggered by changes at or distal to the neuromuscular junction (Ament and Verkerke, 2009). Depletion of glycemic level and tissue glycogen content with the accumulation of lactic acid and ammonia (metabolic products) are also important factors for peripheral fatigue. Continue the supply of energy resources and elimination of metabolic products can recover the peripheral fatigue (Finsterer, 2012). As a consequence, regular exercise with a balanced diet can reduce fatigue (Schwartz, 1999). Wide investigation on natural products has been proved to improve athletic ability and reduce or eliminates fatigue without side effects (Kim et al., 2002).

Tylorrhynchus heterochaetus Quatrefages (Hechong in Chinese) belongs to the family of Nereididae (Fig. 1). T. heterochaetus is commonly found in the Southern, Eastern, and Yellow Sea estuary areas of China. They mainly grow in the paddy field and eat crop roots as their food. The farmers in the Guangdong Province in China use hechong as their daily food because of its high nutritional value (Tang et al., 2009). It has a high protein content of 60% of dry weight with balanced amino acids. The high protein content of T. heterochaetus is because of the big amount of gamete in their body cavities. When it enters the mature stage, their body cavities are occupied by a gamete, which contains protein as the main component (Osanai, 1978). It has also been emphasized in traditional Chinese medicine (TCM) for treating various diseases. According to the theory of the TCM, T. heterochaetus is described as cold, sweet, salty, clearing the lungs tonic and blood aphrodisiac efficacy (Chen et al., 2015). It contains a sufficient amount of fibrinoclase, fibrinogen activator, and collagenase, which could prevent and treat cerebral thrombosis and myocardial infarction. Dried T. heterochaetus is used as the main drug for curing beriberi. People with a deficiency in yin and sweat at nights are suggested to eat T. heterochaetus to reduce the symptoms by TCM doctors. The secretion of milk in parturition mothers can be increased by T. heterochaetus. T. heterochaetus soup with ginger and pork can reduce neurasthenia and pediatric spleen embolism. The boiled T. heterochaetus is used to prevent nocturia due to kidney deficiency and also can promote the prostate function in males. According to some researches, T. heterochaetus contain 10-20 kinds of fatty acids, which are effective to reduce the risk of dyslipidemia and atherosclerosis (Suzuki and Gotoh, 1986).

To date, the anti-fatigue activity of T. heterochaetus has not been assessed and reported yet. Thus, we aim to assess the anti-fatigue effect of T. heterochaetus and to discover the primary mechanism of action.

Section snippets

Preparation of T. heterochaetus aqueous extract

T. heterochaetus was collected from Estuarine Fisheries Research Institute of Doumen, Zhuhai, China in March 2019. The edible part of T. heterochaetus (200 g) was homogenized using a blender for 30 s, and boiled in water for 1 h. The extract was filtered and centrifuged under 3000 rpm for 10 min. The supernatant was separated and stored under −80 °C. The extract was freeze-dried and used for the experiments. The yield of freeze-dried powder was 4.12% w/v.

Experimental animals

Healthy Kunming mice (weight 20 ± 2 g,

Effect of T. heterochaetus extract on body weight

The final day bodyweight of mice was evaluated after 28 days of treatment with low, medium and high doses of T. heterochaetus extract. In that, all the three doses T. heterochaetus extract reduced the body weight significantly (p ≤ 0.05) when compared to the normal control group mice (Fig. 3).

Effect of T. heterochaetus extract on forced running test

As shown in Fig. 4a in the forced running test, all doses of T. heterochaetus extract treated mice showed high running time which was higher significantly (p ≤ 0.05) than the running time of normal group

Discussion

Herein we present the first report on the anti-fatigue effect of T. heterochaetus aqueous extract evaluated by rotating rod, forced running and weight-loaded swimming tests. Weight-loaded swimming test is of one the commonly used method for screening the anti-fatigue effect in animal models (Tang et al., 2008; You et al., 2012). In the test, the degree of fatigue has been assessed depends on the period of swimming time to fatigue (Tanaka et al., 2003). In our investigation, T. heterochaetus

Conclusions

In this study, our combined results proved the anti-fatigue effect of T. heterochaetus aqueous extract. Among the doses, the medium dose (5.41 mg/0.5 mL/20 g) could increase the weight-loaded swimming time of mice by increasing the LDH activity and liver glycogen levels and by delaying BUN and LA accumulation. Moreover, T. heterochaetus increased the capacity of endurance by acting through the AMPK-linked pathway. Hence, T. heterochaetus could be used as a novel natural agent from a food source

Funding

This study is jointly supported by two research grants R201714 and R201914 from Beijing Normal University-Hong Kong Baptist University United International College.

CRediT authorship contribution statement

Ziqing Yang: Data curation, Writing - original draft, Visualization, Investigation. Christudas Sunil: Data curation, Writing - original draft, Visualization, Investigation. Muthukumaran Jayachandran: Data curation, Writing - original draft, Visualization, Investigation, Software, Validation. Xuchu Zheng: Visualization, Investigation. Kuopeng Cui: Conceptualization, Methodology, Software, Supervision, Project administration, Funding acquisition. Yuepeng Su: Conceptualization, Methodology,

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

References (33)

O.V. Bogdanova et al.
Factors influencing behavior in the forced swim test
Physiol. Behav.
(2013)
M.M. Bradford
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding
Anal. Biochem.
(1976)
N.V. Carroll et al.
The determination of glycogen in liver and muscle by use of anthrone reagent
J. Biol. Chem.
(1956)
R.R. Klosterhoff et al.
Anti-fatigue activity of an Arabian-rich pectin from acerola (Malpighia emarginata)
Int. J. Biol. Macromol.
(2018)
R. Kumar et al.
Cordyceps sinensis promotes exercise endurance capacity of rats by activating skeletal muscle metabolic regulators
J. Ethnopharmacol.
(2011)
X. Li et al.
Analysis of chemical components of shiitake polysaccharides and its anti-fatigue effect under vibration
Int. J. Biol. Macromol.
(2009)
Y. Lin et al.
Anti-fatigue and vasoprotective effects of quercetin-3-O-gentiobiose on oxidative stress and vascular endothelial dysfunction induced by endurance swimming in rats
Food Chem. Toxicol.
(2014)
Y. Liu et al.
Antifatigue and increasing exercise performance of Actinida arguta crude alkaloids in mice
J. Food Drug Anal.
(2016)
T. Suzuki et al.
The complete amino acid sequence of giant multisubunit hemoglobin from the polychaete Tylorrhynchus heterochaetus
J. Biol. Chem.
(1986)
W. Tan et al.
Anti-fatigue activity of polysaccharides extract from radix rehmanniae preparata
Int. J. Biol. Macromol.
(2012)

M. Tanaka et al.

Establishment and assessment of a rat model of fatigue

Neurosci. Lett.

(2003)

H. Westerblad et al.

Skeletal muscle: energy metabolism, fiber types, fatigue and adaptability

Exp. Cell Res.

(2010)

H.P. Zhao et al.

Acute toxicity and anti-fatigue activity of polysaccharide-rich extract from corn silk

Biomed. Pharmacother.

(2017)

O.P. Adams

The impact of brief high-intensity exercise on blood glucose levels

Diabetes, Metab. Syndrome Obes. Targets Ther.

(2013)

W. Ament et al.

Exercise and fatigue

Sport. Med.

(2009)

Y.M. Chen et al.

Fucoidan supplementation improves exercise performance and exhibits anti-fatigue action in mice

Nutrients

(2015)

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Anti-fatigue effect of aqueous extract of Hechong (Tylorrhynchus heterochaetus) via AMPK linked pathway

Highlights

Abstract

Graphical abstract

Introduction

Section snippets

Preparation of T. heterochaetus aqueous extract

Experimental animals

Effect of T. heterochaetus extract on body weight

Effect of T. heterochaetus extract on forced running test

Discussion

Conclusions

Funding

CRediT authorship contribution statement

Declaration of competing interest

Physiol. Behav.

Anal. Biochem.

J. Biol. Chem.

Int. J. Biol. Macromol.

J. Ethnopharmacol.

Int. J. Biol. Macromol.

Food Chem. Toxicol.

J. Food Drug Anal.

J. Biol. Chem.

Int. J. Biol. Macromol.

Neurosci. Lett.

Exp. Cell Res.

Biomed. Pharmacother.

The impact of brief high-intensity exercise on blood glucose levels

Diabetes, Metab. Syndrome Obes. Targets Ther.

Exercise and fatigue

Sport. Med.

Fucoidan supplementation improves exercise performance and exhibits anti-fatigue action in mice

Nutrients