Journal of Nanomedicine &Biotherapeutic Discovery

tic Discovpeu
raf Nanomal oerne & Biothcinedi

ISSN: 2155-983X

Suzuki, J Nanomedine Biotherapeutic Discov 2017,
7:2
DOI: 10.4172/2155-983X.1000156

y
er

Jou

Short Communication

Open AccessExhaustive Exercise-Induced Neutrophil-Associated Tissue Damage andPossibility of its Prevention
Suzuki K*
Faculty of Sport Sciences, Waseda University, Tokorozawa, Saitama, Japan*Corresponding

author: Katsuhiko Suzuki, Faculty of Sport Sciences, Waseda University, 2-579-15 Mikajima, Tokorozawa, Saitama 359-1192, Japan, E-mail:katsu.suzu@waseda.jp
Received date: Nov 03, 2017; Accepted date: Dec 01, 2017; Published date: Dec 05, 2017

Copyright: © 2017 Suzuki K. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use,
distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract
Neutrophils and inflammatory cytokines that accumulate in tissues often cause organ damage/dysfunctions.
Herein, background information on some research findings of exertional effects on systemic inflammation centeredon neutrophils is introduced. Furthermore, a functional activity measurement system of neutrophils including cell
migration and reactive oxygen species (ROS) production, and preventive countermeasures against neutrophilinduced pathogenesis (e.g., polyphenols) are described.

Keywords: Exercise; Neutrophil; Oxidative stress; Muscle damage

Introduction
In the blood circulation, there are five kinds of white blood cells(leukocytes), of which neutrophils (polymorphonuclear leukocytes) are
the most abundant. Neutrophils not only play an important role inhost defense against infectious microbes by migrating to the site of
infection and producing reactive oxygen species (ROS) to kill invading
microorganisms. They also mediate pathological processes and acuteinflammatory tissue damage [1]. Migration of neutrophils to the tissue
is the first step to induce local inflammation, and the overproductionof ROS by neutrophils leads to oxidative stress. Although neutrophils
migrate into damaged tissues following injury, infiltration ofneutrophils into damaged muscles within several hours after exercise
has also been demonstrated [2]. In the exercise and sport science fields,
we have reported that exhaustive exercise facilitates neutrophil activity,
suggesting their involvement with muscle damage [3-12]. However,endogenous antioxidant capacity also increases following exercise,
which can partially attenuate neutrophil activation and oxidative stress[13-16]. Because we observed complex phenomena centered on
neutrophils following exercise, some novel technology was required to
examine the neutrophil dynamics and functional modulation.

Development of an Assessment Methodology
Since antioxidants are considered as one of the countermeasuresagainst oxidative stress, it is necessary to develop an assessment
methodology which replicates in vivo conditions more closely. Under
these conditions, whether migrated neutrophils and ROS production
are useful or not for the body can be delineated, and appropriate
countermeasures against inflammation and oxidative stress can beproposed. Therefore, we developed a neutrophil activity measurement
system that analyzes the migratory activity of neutrophils and their
ROS production. In brief, this system involves layering mixed whole
blood and luminol as a chemiluminogenic probe on transparent
hydrogel to detect ROS by luminol-dependent chemiluminescence.
This assay largely monitors myeloperoxidase (MPO)-dependent
formation of highly toxic ROS, such as hypochlorous acid (HOCl).

J Nanomedine Biotherapeutic Discov, an open access journal
ISSN: 2155-983X

Also, the cell count in the hydrogel can be quantified as a measure of
the migratory activity of neutrophils. This new method can be applied
not only for assessing the state of inflammation and oxidative stress ex
vivo, but also as a screening system for predicting the effectiveness ofantioxidant and anti-inflammatory substances in vitro [17-20].

Application of the Experimental ApproachThis methodology identified neutrophil activation ex vivo in theabsence of changes in many cytokines and inflammatory markers after
eccentric muscle-damaging contractions, highlighting the importanceof neutrophil dynamics in the pathological process for exerciseinduced muscle damage. Furthermore, we have demonstrated thatenhanced neutrophil activity after intensive endurance exercise isassociated with muscle and renal damage, which are observed inendurance athletes.
Many antioxidants such as plant extracts, especially polyphenols,
have been screened in vitro for the prevention of lifestyle-relateddiseases, neoplastic diseases, as well as exercise-induced oxidative
stress [17-19]. As a consequence, curcumin, a type of dietarypolyphenol, displayed the most potent inhibitory action on neutrophil
migration and ROS production among the tested antioxidants in vitro[19]. An additional plant extract, Tabebuia avellanedae (taheebo)
extract also displayed inhibitory effects on neutrophil-related oxidative
stress ex vivo [21].

Related and Supplementary Findings
Apart from the above methodology, we have also investigated therole of neutrophils in the exercise-induced muscle damage. We
demonstrated that neutrophils firstly migrate to the damaged muscle,
attract macrophages, and induce inflammatory mediators andcytokines [12]. Although curcumin is known to protect against
ischemia/reperfusion injury in rat skeletal muscle [22], we
demonstrated exercise-induced oxidative stress was attenuated by priorcurcumin ingestion in vivo in humans [23]. Taheebo extract includes
polyphenols that were found to suppress not only oxidative stress, butalso the production of inflammatory cytokines and prostaglandin E2
by blocking cyclooxygenase-2 (COX-2) in the same manner as anti-

Volume 7 • Issue 2 • 1000156

Citation:

Suzuki K (2017) Exhaustive Exercise-Induced Neutrophil-Associated Tissue Damage and Possibility of its Prevention. J Nanomedine
Biotherapeutic Discov 7: 156. doi:10.4172/2155-983X.1000156

Page 2 of 3
inflammatory and pain-relieving drugs such as celecoxib [24,25]. Also,
pretreatment with fucoidan, a known leukocyte-adhesion inhibitor, isreported to reduce muscle hyperalgesia induced by local
administration of P2X3 agonist in a rat model [26]. This indicates thatleukocyte adhesion and subsequent migration results in pain andinflammation, and that leukocyte adhesion and migration are thepoint of action to prevent inflammation and tissue damage.
Aside from functional foods, reducing exercise-induced
dehydration by sports drink ingestion also inhibited neutrophil
activation and cytokine release in vivo [27,28]. Further studies areneeded to investigate which food and fluid ingredients andsupplementation strategies are most effective to reduce inflammation,
because severe systemic inflammation can cause multiple organ failureand heat stroke [29,30]. Therefore, these potential countermeasures
not only help to reduce pathophysiological processes, but also lead to
new research findings for the prevention of oxidative stress,
inflammation, organ damage and dysfunction [31-35].

Concluding Remarks
Strenuous exercise induces leukocytosis mainly due to neutrophilia
in the systemic circulation, while neutrophil activity is associated withskeletal muscle damage and other internal organ dysfunctions. As forthe underlying mechanisms, the research findings of exertional effects
on systemic inflammation centered on neutrophils are describedabove, which are (together with cytokines) in line with thepathogenesis of multiple organ failure in systemic inflammation, heatstroke and sepsis. Herein, the benefits of measuring neutrophil
functional activity, including cell migration and ROS production, andpossible preventive countermeasures targeting pathogenesis have been
described. Future studies are required to examine the validity of suchprevention and treatment approaches, based on their mechanisms ofaction.

5.
6.

7.

8.
9.
10.
11.

12.
13.
14.
15.
16.

Acknowledgements
These works were partly supported by a Grant-in-Aid for the
Scientific Research (A) and Strategic Research Foundation at Private
Universities from the Ministry of Education, Culture, Sports, Scienceand Technology of Japan. I gratefully appreciate my colleagues andlaboratory team for research development and progress. Also, I would
like to thank Dr. Llion Roberts, Lecturer of Human Physiology, Schoolof Allied Health Sciences & Menzies Health Institute Queensland,
Griffith University, and Dr. Jonathan Peake, President of theInternational Society of Exercise and Immunology (ISEI) (http://
www.isei.dk/index.php?pageid=14), for English editing.

17.
18.
19.

20.

References
1.
2.
3.
4.

Smith JA (1994) Neutrophils, host defense, and Inflammation: a doubleedge sword. J Leukocyte Biol 56: 672-686.
Pizza FX, Peterson JM, Baas JH, Koh TJ (2005) Neutrophils contribute tomuscle injury and impair its resolution after lengthening contractions in
mice. J Physiol 562: 899-913.
Suzuki K, Sato H, Kikuchi T, Abe T, Nakaji S, et al. (1996) Capacity ofcirculating neutrophils to produce reactive oxygen species after
exhaustive exercise. J Appl Physiol 81: 1213-1222.
Suzuki K, Naganuma S, Totsuka M, Suzuki KJ, Mochizuki M, et al (1996)
Effects of exhaustive endurance exercise and its one-week daily repetition
on neutrophil count and functional status in untrained men. Int J Sports
Med 17: 205-212.

J Nanomedine Biotherapeutic Discov, an open access journal
ISSN: 2155-983X

21.

22.
23.
24.

Suzuki K, Totsuka M, Nakaji S, Yamada M, Kudoh S, et al. (1999)
Endurance exercise causes interaction among stress hormones, cytokines,
neutrophil dynamics, and muscle damage. J Appl Physiol 87: 1360-1367.
Suzuki K, Yamada M, Kurakake S, Okamura N, Yamaya K, et al. (2000)
Circulating cytokines and hormones with immunosuppressive but
neutrophil-priming potentials rise after endurance exercise in humans.
Eur J Appl Physiol 81: 281-287.
Peake J, Wilson G, Hordern M, Suzuki K, Yamaya K, et al. (2004) Changes
in neutrophil surface receptor expression, degranulation, and respiratory
burst activity after moderate- and high-intensity exercise. J Appl Physiol
97: 612-618.
Peake JM, Suzuki K, Wilson G, Hordern M, Nosaka K, et al. (2005)
Exercise-induced muscle damage, plasma cytokines, and markers ofneutrophil activation. Med Sci Sports Exerc 37: 737-745.
Suzuki K, Peake J, Nosaka K, Okutsu M, Abbiss CR, et al. (2006) Changes
in markers of muscle damage, inflammation and HSP70 after an Ironman
triathlon race. Eur J Appl Physiol 98: 525-534.
Sugama K, Suzuki K, Yoshitani K, Shiraishi K, Kometani T. (2012) IL -17,
neutrophil activation and muscle damage following endurance exercise.
Exerc Immunol Rev 18: 116-127.
Kanda K, Sugama K, Hayashida H, Sakuma J, Kawakami Y, et al. (2013)
Eccentric exercise-induced delayed-onset muscle soreness and changes inmarkers of muscle damage and inflammation. Exer Immunol Rev 19:
74-87.
Kawanishi N, Mizokami T, Niihara H, Yada K, Suzuki K. (2016)
Neutrophil depletion attenuates muscle injury after exhaustive exercise.
Med Sci Sports Exerc 48: 1917-1924.
Suzuki K, Nakaji S, Yamada M, Liu Q, Kurakake S, et al.(2003) Impact of
a competitive marathon race on systemic cytokine and neutrophil
responses. Med Sci Sports Exerc 35: 348-355.
Peake J, Suzuki K. (2004) Neutrophil activation, antioxidant supplements
and exercise-induced oxidative stress. Exerc Immunol Rev 10: 129-141.
Peake JM, Suzuki K, Coombes JS (2007) The influence of antioxidant
supplementation on markers of inflammation and the relationship tooxidative stress after exercise. J Nutri Biochem 18: 357-371.
Sugama K, Suzuki K, Yoshitani K, Shiraishi K, Miura S, et al. (2015)
Changes of thioredoxin, oxidative stress markers, inflammation andmuscle/renal damage following intensive endurance exercise. Exerc
Immunol Rev 21: 130-142.
Suzuki K, Ohno S, Suzuki Y, Ohno Y, Okuyama R, et al. (2012) Effect ofgreen tea extract on reactive oxygen species produced by neutrophils
from cancer patients. Anticancer Res 32: 2369-2375.
Suzuki K, Komaba Y, Tomari M, Suzuki Y, Sugama K, et al. (2012)
Functional assessment of plant extracts by application of novel neutrophil
activity measurement system. Jpn J Complement Altern Med 9: 89-95.
Suzuki K, Tomari M, Takahashi M, Sugama K, Otsuka Y, et al. (2012)
Assessment of antioxidant and anti-inflammatory actions of curcumin by
application of novel neutrophil activity measurement system. Jpn J Clin
Chem 41: 343-348.
Suzuki Y, Ohno S, Okuyama R, Aruga A, Yamamoto M, et al. (2012)
Suzuki K. Determination of chronic inflammatory states in cancer
patients using assay of reactive oxygen species production by neutrophils.
Anticancer Res 32: 565-570.
Ohno S, Ohno Y, Suzuki Y, Miura S, Yoshioka H, et al. (2015) Ingestion of
Tabebuia avellanedae (Taheebo) inhibits production of reactive oxygen
species from human peripheral blood neutrophils. Int J Food Sci Nutr
Diet 6: 1-4.
Avci G, Kadioglu H, Sehirli AO, Bozkurt S, Guclu O, et al. (2012)
Curcumin protects against ischemia/reperfusion in rat skeletal muscle. J
Surg Res 172: 9-46.
Takahashi M, Suzuki K, Kim HK, Otsuka Y, Imaizumi A, et al. (2014)
Effects of curcumin supplementation on exercise-induced oxidative stressin humans. Int J Sports Med 35: 469-475.
Suo M, Isao H, Kato H, Takano F, Ohta T. (2012) Anti-inflammatory
constituents from Tabebuia avellanedae. Fitoterapia 83: 1484-1488.

Volume 7 • Issue 2 • 1000156

Citation:

Suzuki K (2017) Exhaustive Exercise-Induced Neutrophil-Associated Tissue Damage and Possibility of its Prevention. J Nanomedine
Biotherapeutic Discov 7: 156. doi:10.4172/2155-983X.1000156

Page 3 of 3
25.
26.
27.
28.

29.
30.

Ma S, Yada K, Lee H, Fukuda Y, Iida A, et al. (2017) Taheebo polyphenols
attenuate FFA-induced inflammation in murine and human macrophagecell lines as inhibitor of COX-2. Frontiers in Nutrition.
Schiavuzzo JG, Teixeira JM, Melo B, da Silva dos Santos DF, Jorge CO, etal. (2015) Muscle hyperalgesia induced by peripheral P2X3 receptors is
modulated by inflammatory mediators. Neuroscience 285: 24-33.
Suzuki K, Hashimoto H, Oh T, Ishijima T, Mitsuda H, et al. (2013) The
effects of sports drink osmolality on fluid intake and immunoendocrine
responses to cycling in hot conditions. J Nutr Sci Vitaminol 59: 206-212.
Suzuki K, Shiraishi K, Yoshitani K, Sugama K, Kometani T. (2014) The
effect of a sports drink based on highly branched cyclic dextrin on
cytokine responses to exhaustive endurance exercise. J Sports Med Phys
Fitness 54: 622-630.
Suzuki K, Nakaji S, Yamada M, Totsuka M, Sato K, et al. (2002) Systemic
inflammatory response to exhaustive exercise: Cytokine kinetics. Exerc
Immunol Rev 8: 6-48.
Lim CL, Suzuki K. (2017) Systemic inflammation mediates the effects ofendotoxemia in the mechanisms of heat stroke. Biol Med (Aligarh) 9:
1000376.

J Nanomedine Biotherapeutic Discov, an open access journal
ISSN: 2155-983X

31.

32.
33.
34.
35.

Suzuki K, Takahashi M, Li CY, Lin SP, Tomari M, et al. (2015) The acuteeffects of green tea and carbohydrate co-ingestion on systemic
inflammation and oxidative stress during sprint cycling. Appl Physiol
Nutr Metabol 40: 997-1003.
Fang SH, Suzuki K, Lim CL, Chung MS, Ku PW, et al. (2016) Associations
between sleep quality and inflammatory markers in patients withschizophrenia. Psychi Res 246:154-160.
Li CY, Suzuki K, Hung YL, Yang MS, Yu CP, et al. (2017) Aloe metabolites
prevent LPS-induced sepsis and inflammatory response by inhibiting
mitogen-activated protein kinase activation. Am J Chin Med 45: 847-861.
Hung YL, Fang SH, Wang SC, Cheng WC, Liu PL, et al. (2017) Corylin
protects LPS-induced sepsis and attenuates LPS-induced inflammatory
response. Sci Rep 7: 46299.
Peake JM, Roberts LA, Figueiredo VC, Egner I, Krog S, et al. (2017) The
effects of cold water immersion and active recovery on inflammation andcell stress responses in human skeletal muscle after resistance exercise. J
Physiol 595: 695-711.

Volume 7 • Issue 2 • 1000156