Rev Bras Fisiol Exerc 2020;19(1):54-64
doi: 10.33233/rbfe.v19i1.3988
REVIEW
Lactate
as an energy substrate and its role in carcinogenesis
Dayana Pimentel de Souza1, Marvyn de Santana do Sacramento1,2,3, Pedro Henrique Silva Santos1,4, Giulliano Gardenghi1,5,6,7,8
1Faculdade do Centro
Oeste Paulista, Bauru/SP, 2Centro Universitário do Instituto Social da Bahia,
Salvador/BA, 3ACTUS CORDIOS Reabilitação Cardiovascular, Respiratória e Metabólica, Salvador/BA, 4Fundação Estatal de Saúde da Família (FESF),
Bahia, 5Hospital Encore (Aparecida de Goiânia), Goiás/GO, 6Centro de Estudos Avançados e Formação Integrada, Goiânia/GO, 7Lifecare/HUGOL - Unidade de terapia Intensiva em Queimados, Goiânia/GO, 8Hospital e maternidade São Cristovão, São Paulo/SP/Brasil
Received
on October 28, 2019, accepted: on January 17, 2020.
Corresponding author: Dayana Pimentel de
Souza, Rua São Félix, 88 Centro 48790-000 Tucano BA
Dayana Pimentel de
Souza: dayk16@hotmail.com
Marvyn de Santana do
Sacramento: marvynsantana@gmail.com
Pedro Henrique Silva
Santos: pedrossantos121@gmail.com.
Giulliano Gardenghi:
ggardenghi@encore.com.br
Abstract
Introduction: Lactate is the product of the degradation of pyruvate produced in the
cytoplasm. For a long time, it was believed that it was produced only in the
presence of hypoxia. Several studies have shown that lactate production depends
on several factors and is not only influenced by the lactic anaerobic system.
In addition, large concentrations of lactate are present in neoplastic cells,
even at rest, a phenomenon known as the Warburg Effect, which can occur due to
the high metabolic rate of tumor cells. Objective: This study aimed to
discuss the physiological aspects involved in the production, metabolism and
signaling of lactate, as well as to demonstrate the new therapeutic results
related to the cancer clinic. Methods: This is a literature review
study. Articles were selected in the languages: Portuguese, English and
Spanish, published between 2000 and 2019, in the databases: MEDline
via Pubmed, Scientific Electronic Library Online (Scielo). The gray literature was verified using Google
Scholar and reference list of selected articles. Results: 43 articles
related to lactate were included. The searches were carried out between July
and December 2019. Conclusion: Lactate is a subtract produced in aerobic
and anaerobic environments, in different exercise intensities. It can be used
as an energy source during and after physical exercise, in addition to acting
on anabolic signals. On the other hand, it can contribute to the maintenance of
an environment that favors carcinogenic proliferation. This thinking has
allowed the creation of new therapies to decrease tissue damage and eradicate
malignant cells.
Keywords: lactic acid; neoplasm; metabolism.
Resumo
Lactato como substrato
energético e a atividade carcinogênica
Introdução: O lactato é o produto
da degradação do piruvato produzido no citoplasma. Durante muito tempo
acreditou- se que ele era produzido apenas na presença de hipóxia. Diversos
estudos demonstraram que a produção do lactato depende de vários fatores, não
sendo apenas influenciado pelo sistema anaeróbio lático. Além disto, grandes
concentrações de lactato estão presentes em células neoplásicas, mesmo em
repouso, fenômeno conhecido como Efeito Warburg, que pode ocorrer por uma alta
taxa metabólica das células tumorais. Objetivo: Este estudo objetivou
discutir a respeito dos aspectos fisiológicos envolvidos na produção,
metabolismo e sinalização do lactato, bem como, demonstrar os novos resultados
terapêuticos relacionados à clínica do câncer. Métodos: Trata-se de um
estudo de revisão da literatura. Foram selecionados artigos nos idiomas:
português, inglês e espanhol, publicados entre 2000 e 2019, nas bases de dados:
MEDline via Pubmed, Scientific Electronic Library
Online (Scielo). A literatura cinzenta foi verificada
por meio do Google acadêmico e lista de referências dos artigos selecionados. Resultados:
Foram incluídos 43 artigos relacionados ao lactato. As buscas foram realizadas
entre julho e dezembro de 2019. Conclusão: O lactato é um subtrato produzido no meio aeróbio e anaeróbio, em
diferentes intensidades de exercício. Pode ser utilizado como fonte de energia
durante e após o exercício físico, além de atuar em sinalizações anabólicas.
Por outro lado, pode contribuir para manutenção de um ambiente favorecedor à
proliferação carcinogênica. Este pensamento tem permitido a criação de novas
terapias na tentativa de diminuir a agressão ao tecido e erradicar células
malignas.
Palavras-chave: ácido lático;
neoplasia; metabolismo.
Lactate is the end product of the degradation
of pyruvate produced in the cytoplasm. Pyruvate can be metabolized in both the
cytoplasm and mitochondria [1]. For a long time it was
believed that lactate was produced only in the presence of hypoxia. However,
this theory has been modified [2]. One of the studies to challenge this
hypothesis was developed by Loenneke et al. [3]
demonstrating that arterial occlusion in 50 and 60% did not increase lactate
production compared to an occlusion of 40%.
The formation of lactate is proportional to the intensity of the effort.
One of the answers to this physiological adaptation is the increase in pyruvate
concentrations during more vigorous physical exercise, as well as higher
concentrations of catecholamines, which, among other functions, stimulate the
activity of the enzyme glycogen phosphorylase and regulatory enzymes of the
glycolytic pathway. When the capacity of pyruvate released in the cytoplasm
exceeds the capacity of the active transport system to the mitochondrial
environment, thus the formation of lactate increases regardless of the presence
of oxygen [1].
The lactate accumulation also follows a response threshold where there
is a dynamic balance between its production and oxidation, when the exercise is
below the anaerobiosis threshold, a considerable increase with bicarbonate
buffering and an exponential increase from the respiratory compensation point,
leading to an accumulation of this metabolic both inside the muscle fiber and
in the bloodstream. This occurs both because of the loss of buffering capacity
and because of the saturation of monocarboxylate transporters (MCTs), which
transport lactate and increase the recruitment of glycolytic motor units [4].
Lactate is used to generate adenosine triphosphate (ATP) in specific
organs, such as heart muscle, liver, kidney, brain, adipose tissue and skeletal
muscle [5]. In addition, large concentrations of lactate are present in
neoplastic cells, even at rest, a phenomenon known as the Warburg Effect, which
can occur due to a high metabolic rate of tumor cells, which are glycolytic.
Lactate is present in all stages related to carcinogenesis, from the
angiogenesis phase to the self-sufficiency of cancer cells, being of
fundamental importance for the understanding of this pathogenic process [6].
This is a literature review study. The searches were carried out between
July and December 2019, with last verification on December 15, 2019. Articles
were selected in the languages: portuguese, english and spanish, published
between 2000 and 2019, in the databases: MEDline via Pubmed, Scientific Electronic Library Online (Scielo). The gray literature was verified using Google
academic and reference list of selected articles.
Search
strategy
Descriptors in Health Sciences (DeCS) or
Medical Subject Headings (MeSH) were used according
to the specific language of the database: Lactate AND Exercise AND Neoplasm,
along with the synonyms of each descriptor.
Energy
metabolisms and lactate production
The energy systems work simultaneously, regardless of the intensity of
the exercise. Although there is no ordering between the energy systems, there
is a predominance of the oxidative system in relation to the others, a fact that
can be explained by a better oxidation of macronutrients and a higher energy
balance in relation to the amount of ATP produced. Despite this aerobic
predominance, when there is a need for faster ATP resynthesis,
the anaerobic contribution tends to increase. These metabolic pathways are
stimulated by the amount of adenosine diphosphate (ADP), inorganic phosphate
(Pi) and the intracelular pH, as well as the
bioavailability of energy substrates [1].
The three energy systems are:
1-
Phosphate system or ATP/CP - Alactic anaerobic,
energetic substrate used: phosphate-creatine;
2-
Anaerobic glycolysis system (Cytosol) - Anaerobic lactic, energetic substrate
used: glucose;
3-
Oxidative System (Mitochondria) - Aerobic, energetic substrates used: glucose,
fatty acids, proteins [7].
In practical terms, the breakdown of glucose for the formation of ATP
and lactate occurs in ten biochemical reactions. This pathway can be
didactically divided into three phases: investment phase, breakage phase and
redox phase. In the investment phase, as the name implies, this is where the
investment of two molecules of ATP occurs. The second phase is where the hexose
will be cleaved, forming two trioses. In the last phase, the energy balance is
recovered, since four molecules of ATP are produced, and when the difference
between what was invested is analyzed, a positive balance of 2 ATPs is
obtained. In addition to the positive balance of ATPs, there is also the
production of two molecules of water (H2O), two molecules of nicotidamine adenine reduced dinucleotide (NADH+H+)
and two molecules of pyruvate. Under exercise conditions, for example, where
the production of ATP needs to be supplemented by anaerobic glycolysis, the
enzyme cytoplasmic lactate dehydrogenase (LDHc) catalyzes
the oxidation of NADH+H+, reducing the pyruvate
molecules, which, when they affect the two atoms and hydrogen, turn into
lactate. This lactate, as already mentioned, can serve as an intracellular
energy substrate or can be released into the extracelular
space [8].
Pelarigo
et al. [9] conducted a study in ten swimmers to assess the maximum
oxygen consumption (VO2max), the predominance of energy metabolism
around intensities of 97.5, 100 and 102.5% at maximum steady state lactate
(MLSS). Initially, they were submitted to an intermittent incremental protocol
until exhaustion to determine the speed corresponding to the individual
anaerobic threshold. Subsequently, each participant performed three to five
swimming sessions at speed rhythms imposed based on the incremental protocol to
achieve the MLSS. Dominance of the aerobic system was observed with increasing
contribution from the anaerobic system, in addition to a rise in lactate
production as the intensity increased. Therefore, even in the presence of
oxygen, there was an increase in the substrate.
Physical
exercise and muscle fatigue
Currently we know that lactate is not an inducer of muscle fatigue. The
physiological mechanisms involved during and after exercise encompass a series
of neuromuscular, biochemical and metabolic factors which will be addressed
shortly [10,11].
In 2002, Westerblad et al. [12] raised
the question of whether the accumulation of lactate or inorganic phosphate is
the main factor that can lead to fatigue. It turns out that in the acidified
environment, caused by the concentration of the energetic substrate inside the
cell, the muscle contraction capacity decreases, because the active enzymes
decrease its activity, and this whole process results in the insufficiency of
maintaining physical effort.
In 2015, another explanation was considered for some special cases of
muscle fatigue during physical exertion. Exhaustion may be associated with
defects in monocarboxylate transporters. This deficiency results in a delayed
extraction of the lactate carrier over the hydrogen ions during high intensity
exercises [13].
Lactate is a marker of metabolic acidosis caused by the accumulation of
hydrogen ions. These ions being the real cause of fatigue. Since the body is
facing metabolic acidosis, hydrogen inhibits the calcium that binds to troponin
C, the enzymes that provide energy work in a reduced way, the muscle
contraction decreases and can reach exhaustion and even interruption of
exercise depending on the intensity level maintained. In order to combat this
event, lactate transports hydrogen ions out of the muscle or transports it into
the mitochondria where ATP resynthesis occurs [14].
Another point to emphasize, is about late muscle pain, it was believed
that the accumulation of the metabolite (lactate) would be considered the main
factor related to pain that develops during subsequent days after
high-intensity physical exercise [15]. The mechanisms that permeate the
generation of pain are associated with the inflammatory process, where there is
migration of macrophages, release of histamines and quinines, as well as
activation of pain receptors by local edema. This acute process is due to
microlesions generated by an increase in training volume in conjunction with a
series of signals that result in repair of muscle damage [16].
Looking at it from another angle, the accumulation of that metabolite
can be beneficial for skeletal muscle, as it contributes to the activation of
the target of the mammalian rapamycin receptor (mTOR). mTOR is closely linked
to an anabolic signaling pathway, in which it regulates, among other things,
protein synthesis, triggered by exercise [3-10]. This anabolic process after
neuromuscular exercise is associated with the activation of satellite cells,
which, after being activated, generate percussive cells that promote the
synthesis of muscle components, increase in the contractile elements of actin
and myosin, in addition to thickening of the myofibrils and formation of new
sarcomeres [17].
Oishi et al. [18] observed that lactate promotes significant
increases in myogenic levels of follistatin protein,
mTOR protein and P70S6K, while decreasing myostatin levels. Follistatin
is found in different tissues of the body and acts as an antagonist of
myostatin, contributing to the process of regeneration and hypertrophy of
skeletal muscle. In controversy, myostatin is involved in the process of
atrophy and inhibition of muscle regeneration [19].
Monocarboxylate
carriers (MCT proteins)
Monocarboxylates
are lactate membrane transporters in many tissues, including
skeletal
muscle, heart muscle, liver, kidney and brain. Each organ has a
specific carrier that depends on pH to cross the substrate through the
plasma
membrane. MCT1 is found in the cardiac muscle, while MCT1 and MCT4 are
found in
the skeletal muscle [20]. The literature demonstrates the existence of
14
isoforms of monocarboxylates, some of which are already well described
as MCT1
and MCT4 [21]. The transport steps are shown in Figure 1.
MCT2 is expressed in liver, kidney, brain, intestine and testis. The
tissue expression of this protein is extremely low in the human body compared
to the expression of MCT1 [22].
Figure
1 - Illustration of the main stages of transport and
degradation of lactate and pyruvate in the type II fiber (adapted
from Gladden, 2004). Hypothetically, these metabolites could be transported to
the mitochondria by means of MCT1. Then the lactate would be converted to
pyruvate using LDH. Thus, there would be a reduction of NAD and FAD via LDH and
the citric acid cycle (CAC), which would later be oxidized in the electron
transport chain (CTE) for the resynthesis of ATP [2]
MCT1 is suitable for transporting lactate into oxidative cells and MCT4 for
exporting lactate from glycolytic cells to the extracellular medium, although
the direction of transport is obviously determined by the driving force of
lactate and H+ [23].
Studies still differ as to the acute effects of exercise on carrier proteins
[21,24]. However, the work conducted by McGinley & Bishop [25] demonstrated
an increase in acid-base transport proteins (MCT1, MCT4) after 4 weeks of high intense interval training (HIIT).
Use
of lactate by different organs
Lactate is used as an energy source in several organs, including
skeletal muscle, whose fibers are predominantly oxidative and is a major
consumer. During physical exercise, the main destinations for lactate are
skeletal and cardiac muscle for its oxidation, and liver, where the
gluconeogenesis process occurs through the Cori cycle [26].
We can consider that this substrate is produced in large quantities in
glycolytic fibers and consumed in oxidative fibers, resulting in a dynamic
balance between its production and its oxidation. This use of lactate as an
energy substrate in oxidative skeletal muscle is one of the hypotheses to
justify the delay in muscle fatigue in long-term exercises [27].
The
cardiac muscle does not export lactate, it only absorbs and produces it
for consumption. MCT1 resorbs the extracellular to intracellular space.
After
uptaked, the lactate is converted to pyruvate again
by the enzyme LDHc that catalyzes this inverse
reaction and goes on to the Krebs cycle to produce ATP. There is a high amount
of MCT1 in the heart, more than in any skeletal muscle. Therefore, we can
consider that there is a large influx of the metabolite present in this organ,
as well as its use as an energy substrate in exercise [5].
The brain is a highly oxidative organ. It uses glucose as an energy
substrate as its primary source, lactate and ketone bodies as alternative
sources, also transported by MCTs. In exercise, this alternative energy source
can be represented by 33% of utilization in the central nervous system, and,
since the use of lactate by nerve cells increases, that of glucose is reduced
by 25% [28].
The neural system plays an important role in balancing the intra and
extracellular environment. Astrocytes produce lactate by anaerobic glycolysis
and transport is performed by the astrocyte-neuron lactate shuttle. In case of
accumulation of this substrate in the intracellular medium, extrusion into the
bloodstream occurs so that they are reabsorbed by neurons for energy production
[28].
As for the renal system, it participates in the glucose homeostasis
process, having the ability to produce and use it. This production is called
gluconeogenesis and occurs during periods of fasting in the proximal renal
tubule, from precursor substrates, such as lactate, glutamine and glycerol. For
gluconeogenesis to happen depends on two factors: the availability of these
circulating precursor substrates and the intra-tubular enzymatic activity. For
circulating substrates to be available, neurohormonal activation by glucagon
takes place, this signaling allows catecholamines to increase their
concentrations to be reabsorbed by the kidney and be metabolized in the Cori
cycle to produce glucose [29].
The summary of this process is given by the transformation of lactate
into pyruvate, then the pyruvate is transformed into oxaloacetate, which is
converted into phosphoenolpyruvate. Subsequently, phosphoenolpyruvate is
modified to fructose-1,6-bisphosphate by enzymes participating in glycolysis, and
converted to fructose-6-phosphate, this molecule is transformed into
glucose-6-phosphate which will be degraded to generate glucose [30].
Regarding
adipose tissue, the experimental study published in Nature points out
that even under aerobic conditions, white adipocytes
are mainly glycolytic, producing and releasing large amounts of
lactate. The
lactate produced is released into the bloodstream and travels to other
tissues,
including the liver, where it is used for gluconeogenesis. The white
adipose
tissue also absorbs glucose when it is found in high concentrations in
the
blood, therefore, there is an assumption that this absorption is
transformed
into lactate to balance hyperglycemia [30].
The
liver is the main organ to make gluconeogenesis and glycogenolysis.
Gluconeogenesis occurs simultaneously with glycogenolysis, however, it
occurs
at a slow speed initially, as glycogen concentrations become scarce in
the
liver, it starts to act quickly. These two processes are stimulated by
the
hormone glucagon whenever there is a decrease in blood glucose.
Glucagon is an
insulin antagonist, and these two hormones regulate anabolic and
catabolic
metabolism. The metabolic process in which the precursor substrates are
transformed into glucose in the Cori cycle, and prevent the body from
suffering from hypoglycemia, either at rest or during physical
exercise [31,32].
New
discoveries and clinical implications
Lactate
and cancer
Cancer cells are highly multiplicable and have
a high metabolic plasticity. They are nourished by glucose and other
alternative sources, such as glutamine, lactate, fatty acid, so that all these
substrates contribute to their mitosis [33].
Although normal cells depend mainly on oxidative phosphorylation for
their permanence, neoplastic cells survive, proliferate and spread in any
environment, they create mechanisms so that the medium is suitable for their
survival [34].
These cells have a high potential for hyperplasia and to maintain this
disorderly growth they need fast pathways of energy source. In 1920, Otto
Warburg discovered that the mutated cells produce lactate even in the presence
of oxygen, this process became known as aerobic glycolysis or the Warburg
Effect, and to try to explain this phenomenon, the scientist raised the
hypothesis that these cells had a defect in mitochondria, which would lead to
decreased aerobic respiration and a dependence on glycolysis. However, recent
studies have found that not all mitochondria are defective, in addition, tumor
cells use oxidative phosphorylation as a complement. Therefore, another explanation
must be considered, but in the current scenario it is still used [35].
Keenan & Chi [36] bring lactate as an alternative source of survival
in cancer cells that are deprived of glucose in different tissues. This
alternative source of supply is determined by the extraction of the substrate
from the extracellular medium by MCT1 for oxidative cells. In mitochondria,
lactate is transformed into pyruvate, then this pyruvate is taken to the Krebs
cycle to form ATP. High levels of lactate are associated with a poor prognosis
and the proliferation of these cells, with invasion of new tissues, which
configures the installation of metastasis.
Tumor cells feed on glucose at high speed and the formed lactate is
transported to the extracellular medium through MTC4, then it is absorbed into
the oxidative medium by MCT1, in which the process described above is repeated
[37].
The lactate produced by cancer cells activates the hypoxia-induced
factor HIF-1α, and this, in the presence of M2 macrophages, will stimulate
the secretion of Arginase (ARG1). ARG1 is an enzyme that promotes the synthesis
of polyamines, which play an important role in cell proliferation, contributing
in this case to tumor growth. Another negative contribution of lactate is that
when activating HIF-1α, it will influence the secretion of VEGFa promoting angiogenesis. In the presence of new
vessels, neoplastic cells will maintain their nutrition continuously,
benefiting their growth and migration to new tissues, in addition to nourishing
distant cells in the absence of oxygen [38]. Based on this, recent evidence
attributes lactate as a tumor evasion factor to the immune system, this
evasion being one of the hallmark of cancer [39].
Pérez-Escuredo et al. [40] describe
MCTs as beneficial for cancerous metabolism, as it contributes to the resynthesis of lactate, through the ease of transporting
it. In this way, malignant cells create mechanisms of survival and adaptation
to different situations. The acidic medium generated by H+ resulting from the
lactate degradation process, contributes to the development of metastasis,
because of the decrease in enzymatic activity, therefore, the facility to evade
new tissues is high.
The inhibition of monocarboxylate transporters is one of the new
therapies under analysis to fight against the cancer. MCTs are present in tumor
cells, mainly the MCT1 isoform that is related to tumor maintenance, while MCT4
is related to progression. This targeted therapy aims to block lactate
transport through drugs to inhibit this alternative source.[41]
Still talking about the inhibition of monocarboxylates, it was
identified in several types of breast cancer that blocking these transporters
reduced tumor aggressiveness [42]. In contrast, in the study by Guan,
Bryniarski and Morris [43], where they evaluated the inhibition of MCT1 in
murine breast cancer 4T1, an ineffectiveness was observed in this specific type
of carcinoma.
Lactate is a subtract produced in aerobic and anaerobic environments, at
different intensities of exercise. It can be used as an energy source during
and after physical exercise, in addition to acting on anabolic signals. On the
other hand, it can contribute to the maintenance of an environment that favors
carcinogenic proliferation. This thinking has allowed the creation of new
therapies to decrease the aggression to the tissue and eradicate malignant
cells, including by inhibiting the action of monocarboxylates, however these
data are not consolidated for all types of cancer.