Myokine

The term myokine refers to cytokines which are secreted by muscle cells. Cytokines are a diverse group of small proteins or proteoglycans. These intercellular communication molecules fulfill very diverse roles in regulation of cellular expression and differentiation whose systemic effects occur at mere picomolar concentrations. Of particular interest are their effects in tissue regeneration and repair, maintenance of healthy bodily functioning, immunomodulation and embryogenesis.

In 2003, Dr. Bente Klarlund Pedersen et al. suggested that cytokines or other peptides that are produced, expressed and released by muscle fibres (usually during contraction) and exert endocrine effects should be classified as myokines.

Myostatin was the first myokine identified. Its discovery occurred in Se-Jin Lee’s laboratory at Johns Hopkins University in 1997. Both aerobic exercise and strength training in humans and animals attenuate myostatin expression and myostatin inactivation seems to potentiate the beneficial effects of endurance exercise on metabolism.

While myostatin was the first muscle-derived peptide to fulfill the criteria for a myokine, the gp130 receptor cytokine IL-6 (Interleukin 6) was the first myokine that was found to be secreted into the blood stream in response to muscle contractions. Aerobic exercise provokes a systemic cytokine response, including, for example, IL-6, IL-1 receptor antagonist (IL-1ra), and IL-10 (Interleukin 10). IL-6 was serendipitously discovered as a myokine because of the observation that it increased in an exponential fashion proportional to the length of exercise and the amount of muscle mass engaged in the exercise. It has been consistently demonstrated that the plasma concentration of IL-6 increases during muscular exercise. This increase is followed by the appearance of IL-1ra and the anti-inflammatory cytokine IL-10. In general, the cytokine response to exercise and sepsis differs with regard to TNF-α. Thus, the cytokine response to exercise is not preceded by an increase in plasma-TNF-α. Following exercise, the basal plasma IL-6 concentration may increase up to 100-fold, but less dramatic increases are more frequent. The exercise-induced increase of plasma IL-6 occurs in an exponential manner and the peak IL-6 level is reached at the end of the exercise or shortly thereafter. It is the combination of mode, intensity, and duration of the exercise that determines the magnitude of the exercise-induced increase of plasma IL-6.

IL-6 had previously been classified as a proinflammatory cytokine. Therefore, it was first thought that the exercise-induced IL-6 response was related to muscle damage. However, it has become evident that eccentric exercise is not associated with a larger increase in plasma IL-6 than exercise involving concentric “nondamaging” muscle contractions. This finding clearly demonstrates that muscle damage is not required to provoke an increase in plasma IL-6 during exercise. As a matter of fact, eccentric exercise may result in a delayed peak and a much slower decrease of plasma IL-6 during recovery.

IL-6, among an increasing number of other recently identified myokines, thus remains an important topic of myokine research. It appears in muscle tissue and in the circulation during exercise at levels up to one hundred times basal rates, as noted, and is seen as having a beneficial impact on health and bodily functioning in most circumstances. P. Munoz-Canoves et al. write: "It appears consistently in the literature that IL-6, produced locally by different cell types, has a positive impact on the proliferative capacity of muscle stem cells. This physiological mechanism functions to provide enough muscle progenitors in situations that require a high number of these cells, such as during the processes of muscle regeneration and hypertrophic growth after an acute stimulus. IL-6 is also the founding member of the myokine family of muscle-produced cytokines. Indeed, muscle-produced IL-6 after repeated contractions also has important autocrine and paracrine benefits, acting as a myokine, in regulating energy metabolism, controlling, for example, metabolic functions and stimulating glucose production. It is important to note that these positive effects of IL-6 and other myokines are normally associated with its transient production and short-term action."

The emerging understanding of skeletal muscle as a secretory organ, and of myokines as mediators of physical fitness through the practice of regular physical exercise (including in particular strength training and aerobic exercise), as well as new awareness of the anti-inflammatory and thus disease prevention aspects of exercise, is thus transforming our broad understanding of the field of health promotion.

In a 2012 overview, Pedersen and Febbraio presented the following in an article abstract: "During the past decade, skeletal muscle has been identified as a secretory organ. Accordingly, we have suggested that cytokines and other peptides that are produced, expressed and released by muscle fibres and exert either autocrine, paracrine or endocrine effects should be classified as myokines. The finding that the muscle secretome consists of several hundred secreted peptides provides a conceptual basis and a whole new paradigm for understanding how muscles communicate with other organs, such as adipose tissue, liver, pancreas, bones and brain. However, some myokines exert their effects within the muscle itself. Thus, myostatin, LIF, IL-6 and IL-7 are involved in muscle hypertrophy and myogenesis, whereas BDNF and IL-6 are involved in AMPK-mediated fat oxidation. IL-6 also appears to have systemic effects on the liver, adipose tissue and the immune system, and mediates crosstalk between intestinal L cells and pancreatic islets. Other myokines include the osteogenic factors IGF-1 and FGF-2; FSTL-1, which improves the endothelial function of the vascular system; and the PGC-1alpha-dependent myokine irisin, which drives brown-fat-like development. Studies in the past few years suggest the existence of yet unidentified factors, secreted from muscle cells, which may influence cancer cell growth and pancreas function. Many proteins produced by skeletal muscle are dependent upon contraction; therefore, physical inactivity probably leads to an altered myokine response, which could provide a potential mechanism for the association between sedentary behaviour and many chronic diseases."

The authors concluded: "In summary, physical inactivity and muscle disuse lead to loss of muscle mass and accumulation of visceral adipose tissue and consequently to the activation of a network of inflammatory pathways, which promote development of insulin resistance, atherosclerosis, neurodegeneration and tumour growth and, thereby, promote the development of a cluster of chronic diseases. By contrast, the finding that muscles produce and release myokines provides a molecular basis for understanding how physical activity could protect against premature mortality.... Given that muscle is the largest organ in the body, the identification of the muscle secretome could set a new agenda for the scientific community. To view skeletal muscle as a secretory organ provides a conceptual basis for understanding how muscles communicate with other organs such as adipose tissue, liver, pancreas, bone and brain. Physical inactivity or muscle disuse potentially leads to an altered or impaired myokine response and/or resistance to the effects of myokines, which explains why lack of physical activity increases the risk of a whole network of diseases, including cardiovascular diseases, T2DM (Type 2 Diabetes Mellitus), cancer and osteoporosis."

Note that interleukin-15 appears to play a significant role in the reduction of visceral (intra-abdominal or interstitial) fat. This myokine effect is of particular importance, as interstitial fat is prone to inflammation, and is thus seen as a primary contributor to the development of inflammatory diseases. Dr. Pedersen writes, "Recently, we demonstrated that IL-15 mRNA levels were upregulated in human skeletal muscle following a bout of strength training, suggesting that IL-15 may accumulate within the muscle as a consequence of regular training. We further demonstrated a negative association in humans between plasma IL-15 concentration and trunk fat mass, but not limb fat mass. In support of this finding, we demonstrated a decrease in visceral fat mass, but not subcutaneous fat mass, when IL-15 was overexpressed in murine muscle. Quinn and colleagues found that elevated circulating levels of IL-15 in mice resulted in significant reductions in body fat and increased bone mineral content, without appreciably affecting lean body mass or levels of other cytokines. Although this model represents an artificial system, the findings lend some support to the idea that IL-15 secretion from muscle tissue may modulate visceral fat mass specifically via an endocrine mechanism."

Brain-derived neurotrophic factor (BDNF) is also a myokine, though BDNF produced by contracting muscle is not released into circulation. Rather, BDNF produced in skeletal muscle appears to enhance the oxidation of fat. However, research has confirmed that skeletal muscle activation through exercise also contributes to an increase in BDNF secretion in the brain. A beneficial effect of BDNF on neuronal function has been noted in multiple studies. Dr. Pedersen writes, "Neurotrophins are a family of structurally related growth factors, including brain-derived neurotrophic factor (BDNF), which exert many of their effects on neurons primarily through Trk receptor tyrosine kinases. Of these, BDNF and its receptor TrkB are most widely and abundantly expressed in the brain. However, recent studies show that BDNF is also expressed in non-neurogenic tissues, including skeletal muscle. BDNF has been shown to regulate neuronal development and to modulate synaptic plasticity. BDNF plays a key role in regulating survival, growth and maintenance of neurons, and BDNF has a bearing on learning and memory. However, BDNF has also been identified as a key component of the hypothalamic pathway that controls body mass and energy homeostasis.

"Most recently, we have shown that BDNF appears to be a major player not only in central metabolic pathways but also as a regulator of metabolism in skeletal muscle. Hippocampal samples from Alzheimer’s disease donors show decreased BDNF expression and individuals with Alzheimer’s disease have low plasma levels of BDNF. Also, patients with major depression have lower levels of serum BDNF than normal control subjects. Other studies suggest that plasma BDNF is a biomarker of impaired memory and general cognitive function in ageing women and a low circulating BDNF level was recently shown to be an independent and robust biomarker of mortality risk in old women. Interestingly, low levels of circulating BDNF are also found in obese individuals and those with type 2 diabetes. In addition, we have demonstrated that there is a cerebral output of BDNF and that this is inhibited during hyperglycaemic clamp conditions in humans. This last finding may explain the concomitant finding of low circulating levels of BDNF in individuals with type 2 diabetes, and the association between low plasma BDNF and the severity of insulin resistance.

"BDNF appears to play a role in both neurobiology and metabolism. Studies have demonstrated that physical exercise may increase circulating BDNF levels in humans. To identify whether the brain is a source of BDNF during exercise, eight volunteers rowed for 4 h while simultaneous blood samples were obtained from the radial artery and the internal jugular vein. To further identify the putative cerebral region(s) responsible for BDNF release, mouse brains were dissected and analysed for BDNF mRNA expression following treadmill exercise. In humans, a BDNF release from the brain was observed at rest and increased 2- to 3-fold during exercise. Both at rest and during exercise, the brain contributed 70–80% of the circulating BDNF, while this contribution decreased following 1 h of recovery. In mice, exercise induced a 3- to 5-fold increase in BDNF mRNA expression in the hippocampus and cortex, peaking 2 h after the termination of exercise. These results suggest that the brain is a major but not the sole contributor to circulating BDNF. Moreover, the importance of the cortex and hippocampus as sources of plasma BDNF becomes even more prominent in the response to exercise.”

With respect to studies of exercise and brain function, a 2010 report is of particular interest. Erickson et al have shown that the volume of the anterior hippocampus increased by 2% in response to aerobic training in a randomized controlled trial with 120 older adults. The authors also summarize several previously-established research findings relating to exercise and brain function: (1) Aerobic exercise training increases grey and white matter volume in the prefrontal cortex of older adults and increases the functioning of key nodes in the executive control network. (2) Greater amounts of physical activity have been associated with sparing of prefrontal and temporal brain regions over a 9-y period, which reduces the risk for cognitive impairment. (3) Hippocampal and medial temporal lobe volumes are larger in higher-fit older adults (larger hippocampal volumes have been demonstrated to mediate improvements in spatial memory). (4) Exercise training increases cerebral blood volume and perfusion of the hippocampus.

Regarding the 2010 study, the authors conclude: "We also demonstrate that increased hippocampal volume is associated with greater serum levels of BDNF, a mediator of neurogenesis in the dentate gyrus. Hippocampal volume declined in the control group, but higher preintervention fitness partially attenuated the decline, suggesting that fitness protects against volume loss. Caudate nucleus and thalamus volumes were unaffected by the intervention. These theoretically important findings indicate that aerobic exercise training is effective at reversing hippocampal volume loss in late adulthood, which is accompanied by improved memory function."

In 2013, Dr. Pedersen summarized her research findings on myokines as follows: "Skeletal muscle is the largest organ in the body. Skeletal muscles are primarily characterized by their mechanical activity required for posture, movement, and breathing, which depends on muscle fiber contractions. However, skeletal muscle is not just a component in our locomotor system. Recent evidence has identified skeletal muscle as a secretory organ. We have suggested that cytokines and other peptides that are produced, expressed, and released by muscle fibers and exert either autocrine, paracrine, or endocrine effects should be classified as 'myokines.' The muscle secretome consists of several hundred secreted peptides. This finding provides a conceptual basis and a whole new paradigm for understanding how muscles communicate with other organs such as adipose tissue, liver, pancreas, bones, and brain. In addition, several myokines exert their effects within the muscle itself. Many proteins produced by skeletal muscle are dependent upon contraction. Therefore, it is likely that myokines may contribute in the mediation of the health benefits of exercise.

All of the publications of the Danish Centre of Inflammation and Metabolism are accessible through this link: http://www.inflammation-metabolism.dk/index.php?pageid=21

Dr. Pedersen presented a talk at a TEDx conference in Copenhagen on September 18, 2012. This 12-minute presentation highlights her primary research findings in a brief overview format for a general audience.