- Open Access
Improved skeletal muscle fatigue resistance in experimental autoimmune myositis mice following high-intensity interval training
Arthritis Research & Therapy volume 24, Article number: 156 (2022)
Muscle weakness and decreased fatigue resistance are key manifestations of systemic autoimmune myopathies (SAMs). We here examined whether high-intensity interval training (HIIT) improves fatigue resistance in the skeletal muscle of experimental autoimmune myositis (EAM) mice, a widely used animal model for SAM.
Female BALB/c mice were randomly assigned to control (CNT) or EAM groups (n = 28 in each group). EAM was induced by immunization with three injections of myosin emulsified in complete Freund’s adjuvant. The plantar flexor (PF) muscles of mice with EAM were exposed to either an acute bout or 4 weeks of HIIT (a total of 14 sessions).
The fatigue resistance of PF muscles was lower in the EAM than in the CNT group (P < 0.05). These changes were associated with decreased activities of citrate synthase and cytochrome c oxidase and increased expression levels of the endoplasmic reticulum stress proteins (glucose-regulated protein 78 and 94, and PKR-like ER kinase) (P < 0.05). HIIT restored all these alterations and increased the peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) and the mitochondrial electron transport chain complexes (I, III, and IV) in the muscles of EAM mice (P < 0.05).
HIIT improves fatigue resistance in a SAM mouse model, and this can be explained by the restoration of mitochondria oxidative capacity via inhibition of the ER stress pathway and PGC-1α-mediated mitochondrial biogenesis.
Patients with systemic autoimmune myopathies (SAMs), including polymyositis (PM), dermatomyositis (DM), and juvenile DM, suffer from muscle weakness and reduced fatigue resistance , which are associated with debility and increased mortality [2, 3]. The decreased endurance exercise capacity in patients with PM/DM is accompanied by a lower maximal oxygen uptake (VO2 max) [2, 4]. Moreover, mitochondrial dysfunction has been observed in the skeletal muscle of PM/DM patients [2, 5, 6], suggesting a low mitochondrial oxidative capacity as an important mechanism contributing to the reduced fatigue resistance in these patients.
The mechanisms that lead to impaired mitochondrial function in SAMs are not fully clarified. Although mitochondrial dysfunction has been shown to be associated with a proinflammatory microenvironment , there is growing evidence showing that other factors are also involved . In this regard, the endoplasmic reticulum (ER) stress pathways are chronically activated in SAMs  and are associated with impaired mitochondrial function [10, 11]. A previous study has suggested a potential mechanistic link between sustained ER stress and mitochondrial dysfunction, mediated by reactive oxygen/nitrogen species generation due to augmented Ca2+ transfer through the mitochondrial-associated ER membrane (MAMs) [8, 11, 12].
Over a period of years, physical exercise was not recommended to patients with SAMs due to fear of exacerbating muscle inflammation. However, since the safety and benefits of exercise in SAM patients were first shown in 1993 [13, 14], exercise training emerged as a non-pharmacological therapy to improve muscle function and prevent disease progression [15,16,17]. For instance, endurance training improved VO2 max and mitochondrial enzyme activities with reduced disease activity and downregulation of genes related to ER stress in PM/DM patients [2, 18]. Following a resistance exercise training program, improved muscle strength and increased VO2 max were seen in patients with PM/DM, and these improvements were accompanied by a reduction in the gene expression associated with inflammation and fibrosis .
A growing body of evidence demonstrates that high-intensity interval training (HIIT) can serve as an effective alternate to traditional endurance training in healthy individuals and diseased populations . Improvements in aerobic capacity are linked to enhanced peripheral oxygen extraction by the skeletal muscle especially after a few weeks of HIIT [21, 22]. By using in vivo neuromuscular electrical stimulation, we recently demonstrated that the HIIT-induced increase in fatigue resistance is larger with high-intensity than with low-intensity contractions in mouse skeletal muscle, and this effect was linked to improved mitochondria content and function .
Peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) is regarded as an important regulator of mitochondrial biogenesis and function . The AMP-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC), Ca2+/calmodulin-dependent protein kinase II (CaMKII), and p38 mitogen-activated protein kinase are well-known modulators of PGC-1α expression in the skeletal muscle [24,25,26,27]. Previous studies have demonstrated that HIIT increases the phosphorylation levels of these signaling molecules and hence increases the expression of PGC-1α [23, 28]. Moreover, activation of AMPK has been shown to inhibit ER stress and inflammation in the skeletal muscle .
One of the widely used animal models for SAMs is the experimental autoimmune myositis (EAM) mouse . EAM is induced by immunization with three injections of myosin emulsified in complete Freund’s adjuvant. Muscle function is impaired at the end of the immunization period where muscular inflammation is already established [31, 32]. Intriguingly, we recently have reported that resistance training starting 1 day after the last immunization inhibits ER stress and restores muscle strength in mice with EAM . In the present study, we tested the following hypotheses: (1) fatigue resistance is decreased in the muscle of mice with EAM due to the decreased mitochondrial oxidative capacity induced by ER stress and (2) HIIT combats these deleterious effects of EAM.
All experimental procedures were approved by the Committee on Animal Experiments of Sapporo Medical University (No. 18-030). Animal care was in accordance with institutional guidelines.
Induction of experimental autoimmune myositis
Female BALB/c mice (8 weeks old, n = 28) and male Wistar rat (9 weeks old, n = 1) were supplied by Sankyo Lab Service (Sapporo, Japan). Mice were given food and water ad libitum and housed in an environmentally controlled room (24 ± 2 °C) with a 12-h light-dark cycle. Health was monitored by weight and general assessment of animal activity (every other day). EAM was induced by immunizing mice with partially purified myosin, including myosin-binding protein C, as reported previously [30, 33]. Briefly, the skeletal muscle (30 g) obtained from a Wistar rat was minced and washed four times in 30 mM KCl/150 mM sodium phosphate buffer (pH 7.5), 1 mM EDTA, and 1 mM DTT. Myosin was extracted by incubation of the muscle sample with 90 ml chilled 300 mM KCl/150 mM phosphate buffer containing 5 mM MgCl2, 5 mM ATP, 1 mM DTT, and 1 mM EDTA on ice for 45 min with constant agitation. The homogenate was centrifuged for 30 min at 4 °C at 2200g. For myosin precipitation, the supernatant was collected, filtered, and diluted with 15 volumes of chilled ultrapure water. The precipitate was recovered via centrifugation for 10 min at 4 °C at 10,000g, dissolved in 500 mM KCl, and stored at – 80 °C. Purified rat myosin (10 mg/ml) was emulsified with an equal amount of complete Freund’s adjuvant (Difco) with 3.3 mg/ml Mycobacterium butyricum (Difco). BALB/c mice were each immunized intracutaneously with 50–100 μl of an emulsion into three to four locations (a total of 200 μl) on the back on days 0, 7, and 14. One hour after the first immunization, pertussis toxin (500 ng in 100 μl saline; List Biological Laboratories) was intraperitoneally injected into each animal. In the present study, all treated animals underwent successful EAM, defined by a significant increase in spleen weight.
To assess the molecular and physiological adaptations induced by HIIT in the skeletal muscle of EAM mice, we performed two separate experiments. The primary outcome of this study will be fatigue resistance. Secondary outcomes constitute mitochondrial enzyme activity, the amount of mitochondrial respiratory complexes and ER stress-related proteins, myosin heavy chain (MyHC) isoforms, and the phosphorylation levels of signaling proteins.
We first examined the effect of HIIT on muscle fatigability and ER/mitochondrial adaptation in EAM mice. Female BALB/c mice (n = 12) were randomly assigned to CNT (n = 6) and EAM (n = 6) groups. Random numbers were generated using the standard = RAND() function in Microsoft Excel. In the EAM group, HIIT was performed on the left leg (referred to as the EAM + HIIT group), and the right leg served as a non-training EAM control. HIIT was started 24 h after the last immunization and was carried out every other day for a total of 14 sessions (Fig. 1A). The training order was randomized daily, with each animal trained at a different time each training day. Under isoflurane anesthesia, mice were placed supine on a platform with the foot secured to a footplate connected to a torque sensor (S-14154, Takei Scientific Instruments) at an angle of 0° dorsiflexion (i.e., 90° relative to the tibia). The plantar flexor muscles were activated by supramaximal (45 V, 0.5 ms) monophasic rectangular current pulses via a pair of surface electrodes. The stimulation scheme was designed to mimic the activation pattern during all-out cycling bouts, i.e., 0.25 s contractions produced every 0.5 s [23, 34]. Each session consisted of six sets of 60 contractions at 4-min intervals. Twenty-four hours after the last HIIT session, in vivo fatigue resistance of the plantar flexor muscles in each group was measured by 80 repeated 350 ms, 70 Hz tetani given at an interval of 3 s. This was done by an investigator unaware of the treatment side. Twenty-four hours after the measurement of fatigue resistance (i.e., 48 h after the last HIIT session), mice were killed by cervical dislocation under isoflurane anesthesia and the gastrocnemius (GAS) and the plantaris muscles were used for skinned muscle fiber experiments and for biochemical analyses (see below).
To investigate cellular signaling that underlies the HIIT-induced physiological adaptations, female BALB/c mice (n = 16) were randomly divided into the same groups as in experiment 1 (n = 8 in each group). Immediately after one HIIT session, mice were killed by rapid cervical dislocation under isoflurane anesthesia, and the muscles were subsequently isolated. The phosphorylation levels of AMPKα Thr172, CaMKII Thr286, ACC Ser79, and p38 MAPK The180/Tyr182 were investigated in the GAS muscles of each animal.
Myosin heavy chain isoforms separation
Aliquots of GAS muscle extracts containing 5 μg protein were used for myosin heavy chain (MyHC) electrophoresis as previously described . Using a 6.8% polyacrylamide slab gel, electrophoresis was run at 4 °C for 24 h at 160 V and stained with Coomassie brilliant blue. Images of gels were densitometrically evaluated with ImageJ.
Measurement of Ca2+-activated force in skinned muscle fibers
Chemically skinned muscle fibers were prepared, and Ca2+-activated force was measured as described previously . The GAS muscle was pinned out at resting length under paraffin oil and kept at 4 °C. Single muscle fibers were dissected under a stereomicroscope. Four to six skinned fibers were obtained from one whole muscle. A segment of the skinned fiber was connected to a force transducer (Muscle Tester, World Precision Instruments) and then incubated with a N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid (HEPES)-buffered solution (see below) containing 1% (vol/vol) Triton X-100 for 10 min in order to remove the membranous structures. Fiber length was adjusted to optimal length (2.5 μm) by laser diffraction as described previously , and the contractile properties were measured at room temperature (24 °C).
All solutions were prepared as described in detail elsewhere . They contained (in mM) 36 Na+, 126 K+, 90 HEPES, 8 ATP, and 10 creatine phosphate and had a pH of 7.09–7.11 and a free Mg2+ concentration set at 1.0 mM. The maximum Ca2+ solution contained 49.5 mM Ca-EGTA and 0.5 mM free EGTA, whereas the relaxation solution contained 50 mM free EGTA. Various pCa (-log free Ca2+ concentration) solutions (pCa 6.4, 6.2, 6.0, 5.8, 5.6, 5.4, and 4.7) were prepared by mixing the maximum Ca2+ solution and the relaxation solution in appropriate proportions . The contractile apparatus was directly activated by exposing the skinned fiber to the various pCa solutions, and force was measured. The isometric force produced at each pCa was expressed as a percentage of the corresponding maximum force and analyzed by fitting a Hill curve using the SigmaPlot 13.0 software to establish the pCa50 (pCa at half-maximum force). The cross-sectional area of fibers was calculated from the measurements of their diameters. The maximum Ca2+-activated force per cross-sectional area (Fmax) is expressed as mN/mm2.
Mitochondrial enzyme activity
The maximal activities of citrate synthase (CS) and cytochrome c oxidase (COX) were determined in whole muscle homogenates. In brief, the whole plantaris muscles were homogenized in ice-cold 100 mM potassium phosphate buffer (100 μl/mg wet wt), and maximal CS and COX activities were measured spectrophotometrically as described previously [40, 41].
Immunoblots were performed as previously described  using anti-PGC-1α (ab54481, Abcam), anti-total OXPHOS rodent WB antibody cocktail (ab110413, Abcam), anti-adenosine monophosphate deaminase 1 (AMPD1, NBP2-24509, Novus Biologicals), anti-dystrophin (ab15277, Abcam), anti-glucose-regulated protein (Grp) 78 (ADI-SPA-826, Enzo Life Sciences), anti-Grp94 (ADI-SPA-851, Enzo Life Sciences), anti-inositol-requiring transmembrane kinase/endoribonuclease 1α (IRE1α) (#3294, Cell Signaling), anti-PKR-like endoplasmic reticulum kinase (PERK) (#5683, Cell Signaling), anti-phospho-AMPKα Thr172 (#2531, Cell Signaling), anti-AMPKα (#2532, Cell Signaling), anti-phospho-CaMKII Thr286 (#12716, Cell Signaling), anti-CaMKII (611292, BD Biosciences, San Jose, CA), anti-phospho-ACC Ser79 (#3661, Cell Signaling), anti-ACC (#3662, Cell Signaling), anti-phospho-p38 MAPK (#4511, Cell Signaling), and anti-p38 MAPK (#9212, Cell Signaling).
Muscle pieces were homogenized in ice-cold homogenizing buffer (40 μl/mg wet wt) consisting of (mM) the following: Tris maleate, 10; NaF, 35; NaVO4, 1; 1% Triton X 100 (vol/vol); and 1 tablet of protease inhibitor cocktail (Roche) per 50 ml. The protein content was determined using the Bradford assay . Aliquots of the whole muscle homogenates (20 μg) were diluted with Laemmli buffer (mM): urea, 4000; Tris/HCl, 250; SDS, 3.5; 20% glycerol (vol/vol); and 0.0005% bromophenol blue (wt/vol). Proteins were applied to a 4–15% Criterion Stain-Free Gel (BioRad). Gels were imaged (BioRad Stain Free imager), and then proteins were transferred onto the polyvinylidene fluoride membranes and were blocked in 3% (wt/vol) non-fat milk and Tris-buffered saline containing 0.05% (vol/vol) Tween 20, followed by incubation with primary antibody overnight at 4 °C. The membranes were then washed and incubated for 1 h at room temperature with secondary antibody (1:5000, donkey anti-rabbit or donkey anti-mouse, BioRad). Images of the membrane were collected following exposure to chemiluminescence substrate (Millipore) using a charge-coupled device camera attached to ChemiDOC MP (BioRad), and the Image Lab software (BioRad) was used for detection as well as densitometry. The levels of protein expression were normalized to the total proteins from the stain-free image.
Data are presented as mean ± SEM. Data normality was examined with the Shapiro-Wilk test. In experiment 1, for normally distributed data (the distribution of the MyHC isoforms, CS activity, COX activity, the expression levels of PGC-1α, NDUFB8, SDHB, UQCRC, MTC01, ATP5, AMPD1, dystrophin, Grp78, Grp94, IRE1α, and PERK, Fmax, pCa50), one-way ANOVA was used to determine the mean differences among the three groups (CNT, EAM, and EAM+IT group). Fatigue resistance (group × repetitions) and specific force-pCa relationship (group × pCa) were assessed by two-way repeated-measures ANOVA. In experiment 2, for normally distributed data (the phosphorylation levels of AMPK, CaMKII, and p38MAPK), one-way ANOVA was used to determine the mean differences between the groups. When these ANOVA tests showed significance, Bonferroni or Tukey post hoc test was performed. If data exhibited a non-normal distribution (the phosphorylation levels of ACC), a Kruskal-Wallis one-way ANOVA was used on ranks. A P value less than 0.05 was regarded as statistically significant. A power test was performed assuming changes in physiological measurements after HIIT being 30 ± 20% of the control value. With a power of 0.80 and an alpha of 0.05, this gives a sample size of six. Based on this, we used 6 and 8 animals in each group in experiments 1 and 2, respectively, but some analyses were performed with n = 5–6 (ER stress proteins). Statistical testing was performed with SigmaPlot (version 13, Systat Software, Inc.).
HIIT improves fatigue resistance in the skeletal muscle of EAM mice
In experiment 1, there was no difference in body weight between CNT (n = 6) and mice with EAM (n= 6) (mean ± SEM 21.4 ± 0.4 g versus 19.1 ± 0.7 g; P > 0.05). In contrast, the spleen weight was 2.6-fold higher in the EAM than in the CNT group (mean ± SEM 289 ± 20 mg versus 110 ± 5 mg, P < 0.05). The GAS muscle weight was 25% lower in the EAM than in the CNT group (mean ± SEM 70.9 ± 2.2 mg versus 94.2 ± 2.6 mg, P < 0.05), and this was not ameliorated by HIIT (mean ± SEM 75.9 ± 3.5 mg, P > 0.05).
Typical torque traces during a HIIT session are shown in Fig. 1B. Note that the torque was decreased much faster in the later sets than in the early sets. Figure 2A–C shows the representative torque records during in vivo fatiguing stimulations of the plantar flexor muscles from CNT and EAM mice with or without HIIT. The EAM muscles were less fatigue resistance than the control muscles (Fig. 2D, P < 0.05). Importantly, HIIT significantly improved fatigue resistance in the EAM muscles (P < 0.05). The differences in fatigue resistance were not due to any changes in muscle fiber type composition (Fig. 2E, F).
Figure 3A shows the typical traces of Ca2+-activated force in skinned fibers from the GAS muscles in each group. The fiber diameter was smaller in the EAM (37.9 ± 0.7 μm [n = 31 fibers], P < 0.05) and the EAM + HIIT (34.9 ± 1.0 μm [n = 31 fibers], P < 0.05) groups than in the CNT group (43.6 ± 1.0 μm [n = 28 fibers]). Ca2+-activated specific force production was lower in skinned fibers from the EAM muscles compared to those from the control muscles (Fig. 3B, P < 0.05). Notably, this was restored by HIIT to the control level (P < 0.05). The Fmax was 21% lower in the EAM muscle fibers than in the CNT muscle fibers (278 ± 11 mN/mm2 versus 349 ± 11 mN/mm2, P < 0.05), which was recovered by HIIT (353 ± 13 mN/mm2, P < 0.05) (Fig. 3C). The Ca2+ sensitivity (pCa50) was similar in the three groups (Fig. 3D, P > 0.05).
HIIT increases mitochondrial respiratory complexes in the skeletal muscle of EAM mice
Compared to the CNT group, CS and COX activities were lower in the EAM group (Fig. 4A, B, P < 0.05). Notably, these EAM-induced deleterious alterations were restored by HIIT (P < 0.05). Moreover, HIIT markedly increased the protein expression of PGC-1α and mitochondrial respiratory complexes I, III, and IV in the EAM group (Fig. 4C–F, P < 0.05).
Although the precise role in physiology is unsettled, a previous study has proposed that a deficiency of AMPD1, a rate-limiting enzyme involved in the catabolism of AMP to IMP and NH3, may contribute to muscle fatigue . The amount of AMPD1 was significantly lower in the EAM group than in the CNT group (Fig. 4G, H, P < 0.05). In contrast, HIIT further reduced the amount of AMPD1 in the EAM muscles (P < 0.05). Additionally, an acquired reduction in dystrophin has been found in patients with SAMs  and a class I major histocompatibility complex (MHC)-transgenic mouse model of SAMs . However, there was no difference in the amount of dystrophin between the groups (Fig. 4G, H, P > 0.05).
HIIT alleviates ER stress in the skeletal muscle of EAM mice
Previous studies suggest that the ER stress pathways are chronically activated and may play an etiological role in SAM . Accordingly, the GAS muscles of EAM mice showed significantly increased expression of the unfolded protein response proteins Grp78, Grp94, and PERK, but not IRE-1α (Fig. 5A–H, P < 0.05). Notably, HIIT attenuated the increased expressions of these ER stress-related proteins.
The phosphorylation levels of signaling proteins are increased after a single bout of HIIT
In experiment 2, the body weight was slightly higher in mice with EAM (n = 8) than in mice with CNT (n = 8) (mean ± SEM 19.4 ± 0.3 g versus 18.0 ± 0.3 g; P < 0.05). The spleen weight was 5-fold higher in the EAM than in the CNT group (mean ± SEM 432 ± 11 mg versus 87 ± 4 mg, P < 0.05). The phosphorylation levels of AMPK Thr172, ACC Ser79, and p38 MAPK Thr180/Tyr182 did not differ between the CNT and the EAM groups, while the phosphorylation levels of these molecules were increased immediately after one HIIT session compared to the CNT group (Fig. 6A, B, D, E, F, P < 0.05). On the other hand, CaMKII Thr286 phosphorylation was higher in the EAM group than in the CNT group (Fig. 6A, C, P < 0.05), which was not affected by an HIIT session.
In accordance with our hypothesis, we show a reduced fatigue resistance during in vivo fatiguing stimulation in the skeletal muscle of EAM mice, an animal model for acute SAMs . The decreased fatigue resistance was accompanied by the increased expression of ER stress-related proteins and reduced activities of mitochondrial oxidative enzymes. Importantly, these deleterious events were restored by HIIT starting 24 h after the last immunization where muscle function is impaired.
Theoretically, the fatigue resistance of muscle fiber depends on the fiber type, which is defined by the MyHC isoform. However, the impaired endurance performance was not accompanied by an alteration in MyHC isoforms in the skeletal muscle of EAM mice, an animal model for acute SAMs. In line with this, untreated newly diagnosed patients with PM/DM had a similar fiber type composition to healthy individuals, although patients with chronic PM/DM display fewer slow-twitch type I fibers . Thus, alterations in muscle fiber types towards more fatigable isoforms likely contribute to the reduced fatigue resistance at chronic stages, but not at disease onset, of SAMs. On the other hand, previous studies suggest a low mitochondrial respiratory capacity as an important mechanism contributing to the impaired endurance performance in patients with SAMs [2, 5, 6]. In agreement, our data show that the reduced fatigue resistance is accompanied by decreased activities of CS and COX in the skeletal muscle of mice with EAM.
The mechanisms underlying the impaired mitochondrial function in SAMs remain uncertain, although non-immune-mediated pathways are thought to be involved. Indeed, despite the recommended treatment with conventional immunosuppressive agents, few SAM patients regain full muscle endurance performance . In this regard, there is growing evidence to suggest that ER stress pathways are chronically activated in SAMs  and are linked to mitochondrial dysfunction [10, 48]. Recently, Thoma et al.  have shown that the ER stress inducer, tunicamycin, promotes mitochondrial dysfunction in a human skeletal muscle cell line. Moreover, it has been demonstrated that PERK, a key ER stress sensor of the unfolded protein response, resides in MAMs and plays a critical role in mitochondrial dysfunction . Accordingly, our findings of the decreased activities of mitochondrial oxidative enzymes in combination with the increased ER stress proteins Grp78, Grp94, and PERK suggested that sustained ER stress underlies mitochondrial dysfunction in the skeletal muscle of EAM mice.
The improvement of fatigue resistance by HIIT in EAM can be explained by increased muscle aerobic capacity as judged by the upregulation of mitochondrial respiratory complexes (I, III, and IV) and increased activities of CS and COX. The increased CS activity and fatigue resistance have also been reported in patients with PM/DM who performed 12 weeks of endurance training [2, 18]. Notably, one HIIT session in the skeletal muscle of EAM mice was followed by phosphorylation of AMPK, ACC, and p38 MAPK, which was associated with increased PGC-1α protein expression after 4 weeks of HIIT. Importantly, we recently have demonstrated that the same protocol done with normal mice also improves resistance to fatigue accompanied by similar molecular changes in PGC-1a and mitochondrial function , indicating that the experimental model is standardized and validated in healthy animals. Accordingly, these data indicate that the PGC-1α-dependent augmentation of mitochondrial oxidative capacity can be effectively induced by exercise training even under inflammatory conditions such as SAMs.
In addition to mitochondrial biogenesis, HIIT may improve muscle aerobic capacity by ameliorating mitochondrial dysfunction due to ER stress in EAM mice. Indeed, our data show that HIIT inhibited the increased amount of ER stress proteins Grp78, Grp94, and PERK in the skeletal muscle of EAM mice. This is in line with a previous study from our lab where EAM-induced upregulation of ER stress proteins, including Grp78 and Grp94, was attenuated by 4 weeks of high-intensity eccentric contraction training in EAM mice . Taken together, our findings promote exercise as an important non-pharmacological approach for relieving ER stress and improving mitochondrial function. Although the mechanisms underlying this beneficial effect of exercise remain unresolved, previous studies suggest that AMPK functions as a suppressor of ER stress [29, 49].
The skeletal muscle of patients with SAMs [50, 51] and a class I MHC-transgenic mouse model of SAMs  exhibit a reduction of AMPD1, which catalyzes the deamination of AMP to IMP and plays an important role in the purine nucleotide cycle. It has been proposed that AMPD1 deficiency is responsible for muscle weakness in a class I MHC-transgenic mouse . Although we also observed a reduction in AMPD1 content in the skeletal muscle of EAM mice, HIIT-induced increase in fatigue resistance was accompanied by decreased rather than increased amount of AMPD1 in those muscles. Several studies of human subjects have reported variable results with some studies suggesting the mutation of the AMPD1 gene may cause easy fatigability while others indicate individuals with this inherited defect are completely asymptomatic . Cheng et al.  have revealed using AMPD1 knockout mice that AMPD1 deficiency results in no abnormality in muscle performance in both sprint and endurance exercise protocols. Thus, these data suggest that an AMPD1 deficiency may not be involved in the mechanism underlying reduced fatigue resistance in patients with SAMs.
There is much debate about which animal model better mimics different aspects of pathology in SAMs. It would be intriguing for a future study to perform the same experiments using an alternative model. For example, a class I MHC-transgenic mouse model of SAMs has recently been shown to exhibit muscle weakness in combination with an acquired reduction in dystrophin  in line with what is seen in many patients . This acquired reduction in dystrophin could possibly lead to increased muscle damage from eccentric contractions or HIIT, in contrast to the EAM model used here, which does not have a reduction in dystrophin.
We here show reduced fatigue resistance in the skeletal muscle of a mouse model of acute SAM. This functional defect was due to decreased mitochondria oxidative capacity, which was at least in part caused by activation of ER stress-dependent pathway. HIIT-mimicking electrical stimulation reversed these alterations and markedly improved fatigue resistance without any signs of deleterious effects on the skeletal muscle. Thus, our findings highlight the clinical importance of HIIT as a safe and effective way to treat increased muscle fatiguability in patients with SAMs.
Availability of data and materials
The dataset supporting the conclusions of this article is included within the article.
Harris-Love MO, Shrader JA, Koziol D, Pahlajani N, Jain M, Smith M, et al. Distribution and severity of weakness among patients with polymyositis, dermatomyositis and juvenile dermatomyositis. Rheumatology (Oxford). 2009;48:134–9.
Alemo Munters L, Dastmalchi M, Katz A, Esbjornsson M, Loell I, Hanna B, et al. Improved exercise performance and increased aerobic capacity after endurance training of patients with stable polymyositis and dermatomyositis. Arthritis Res Ther. 2013;15:R83.
Dobloug GC, Svensson J, Lundberg IE, Holmqvist M. Mortality in idiopathic inflammatory myopathy: results from a Swedish nationwide population-based cohort study. Ann Rheum Dis. 2018;77:40–7.
Wiesinger GF, Quittan M, Nuhr M, Volc-Platzer B, Ebenbichler G, Zehetgruber M, et al. Aerobic capacity in adult dermatomyositis/polymyositis patients and healthy controls. Arch Phys Med Rehabil. 2000;81:1–5.
Campos Y, Arenas J, Cabello A, Gomez-Reino JJ. Respiratory chain enzyme defects in patients with idiopathic inflammatory myopathy. Ann Rheum Dis. 1995;54:491–3.
Meyer A, Laverny G, Allenbach Y, Grelet E, Ueberschlag V, Echaniz-Laguna A, et al. IFN-beta-induced reactive oxygen species and mitochondrial damage contribute to muscle impairment and inflammation maintenance in dermatomyositis. Acta Neuropathol. 2017;134:655–66.
Harty LC, Biniecka M, O’Sullivan J, Fox E, Mulhall K, Veale DJ, et al. Mitochondrial mutagenesis correlates with the local inflammatory environment in arthritis. Ann Rheum Dis. 2012;71:582–8.
Lightfoot AP, McArdle A, Jackson MJ, Cooper RG. In the idiopathic inflammatory myopathies (IIM), do reactive oxygen species (ROS) contribute to muscle weakness? Ann Rheum Dis. 2015;74:1340–6.
Nagaraju K, Casciola-Rosen L, Lundberg I, Rawat R, Cutting S, Thapliyal R, et al. Activation of the endoplasmic reticulum stress response in autoimmune myositis: potential role in muscle fiber damage and dysfunction. Arthritis Rheum. 2005;52:1824–35.
Cao SS, Kaufman RJ. Endoplasmic reticulum stress and oxidative stress in cell fate decision and human disease. Antioxid Redox Signal. 2014;21:396–413.
Thoma A, Lyon M, Al-Shanti N, Nye GA, Cooper RG, Lightfoot AP. Eukarion-134 attenuates endoplasmic reticulum stress-induced mitochondrial dysfunction in human skeletal muscle cells. Antioxidants (Basel). 2020;9:710.
Verfaillie T, Rubio N, Garg AD, Bultynck G, Rizzuto R, Decuypere JP, et al. PERK is required at the ER-mitochondrial contact sites to convey apoptosis after ROS-based ER stress. Cell Death Differ. 2012;19:1880–91.
Escalante A, Miller L, Beardmore TD. Resistive exercise in the rehabilitation of polymyositis/dermatomyositis. J Rheumatol. 1993;20:1340–4.
Hicks JE, Miller F, Plotz P, Chen TH, Gerber L. Isometric exercise increases strength and does not produce sustained creatinine phosphokinase increases in a patient with polymyositis. J Rheumatol. 1993;20:1399–401.
Alemo Munters L, Alexanderson H, Crofford LJ, Lundberg IE. New insights into the benefits of exercise for muscle health in patients with idiopathic inflammatory myositis. Curr Rheumatol Rep. 2014;16:429.
de Oliveira DS, Misse RG, Lima FR, Shinjo SK. Physical exercise among patients with systemic autoimmune myopathies. Adv Rheumatol. 2018;58:5.
Nader GA, Lundberg IE. Exercise as an anti-inflammatory intervention to combat inflammatory diseases of muscle. Curr Opin Rheumatol. 2009;21:599–603.
Alemo Munters L, Loell I, Ossipova E, Raouf J, Dastmalchi M, Lindroos E, et al. Endurance exercise improves molecular pathways of aerobic metabolism in patients with myositis. Arthritis Rheumatol. 2016;68:1738–50.
Nader GA, Dastmalchi M, Alexanderson H, Grundtman C, Gernapudi R, Esbjörnsson M, et al. A longitudinal, integrated, clinical, histological and mRNA profiling study of resistance exercise in myositis. Mol Med. 2010;16:455–64.
Gibala MJ, Little JP, Macdonald MJ, Hawley JA. Physiological adaptations to low-volume, high-intensity interval training in health and disease. J Physiol. 2012;590:1077–84.
Jacobs RA, Flück D, Bonne TC, Bürgi S, Christensen PM, Toigo M, et al. Improvements in exercise performance with high-intensity interval training coincide with an increase in skeletal muscle mitochondrial content and function. J Appl Physiol (1985). 2013;115:785–93.
Raleigh JP, Giles MD, Islam H, Nelms M, Bentley RF, Jones JH, et al. Contribution of central and peripheral adaptations to changes in maximal oxygen uptake following 4 weeks of sprint interval training. Appl Physiol Nutr Metab. 2018;43:1059–68.
Yamada T, Kimura I, Ashida Y, Tamai K, Fusagawa H, Tohse N, et al. Larger improvements in fatigue resistance and mitochondrial function with high- than with low-intensity contractions during interval training of mouse skeletal muscle. FASEB J. 2021;35:e21988.
Egan B, Zierath JR. Exercise metabolism and the molecular regulation of skeletal muscle adaptation. Cell Metab. 2013;17:162–84.
Wojtaszewski JF, Nielsen P, Hansen BF, Richter EA, Kiens B. Isoform-specific and exercise intensity-dependent activation of 5′-AMP-activated protein kinase in human skeletal muscle. J Physiol. 2000;528(Pt 1):221–6.
Rose AJ, Kiens B, Richter EA. Ca2+-calmodulin-dependent protein kinase expression and signalling in skeletal muscle during exercise. J Physiol. 2006;574:889–903.
Akimoto T, Pohnert SC, Li P, Zhang M, Gumbs C, Rosenberg PB, et al. Exercise stimulates Pgc-1alpha transcription in skeletal muscle through activation of the p38 MAPK pathway. J Biol Chem. 2005;280:19587–93.
Fiorenza M, Gunnarsson TP, Hostrup M, Iaia FM, Schena F, Pilegaard H, et al. Metabolic stress-dependent regulation of the mitochondrial biogenic molecular response to high-intensity exercise in human skeletal muscle. J Physiol. 2018;596:2823–40.
Salvadó L, Coll T, Gómez-Foix AM, Salmerón E, Barroso E, Palomer X, et al. Oleate prevents saturated-fatty-acid-induced ER stress, inflammation and insulin resistance in skeletal muscle cells through an AMPK-dependent mechanism. Diabetologia. 2013;56:1372–82.
Allenbach Y, Solly S, Gregoire S, Dubourg O, Salomon B, Butler-Browne G, et al. Role of regulatory T cells in a new mouse model of experimental autoimmune myositis. Am J Pathol. 2009;174:989–98.
Himori K, Ashida Y, Tatebayashi D, Abe M, Saito Y, Chikenji T, et al. Eccentric resistance training ameliorates muscle weakness in a mouse model of idiopathic inflammatory myopathies. Arthritis Rheumatol. 2021;73(5):848–57.
Prevel N, Allenbach Y, Klatzmann D, Salomon B, Benveniste O. Beneficial role of rapamycin in experimental autoimmune myositis. PLoS One. 2013;8:e74450.
Suzuki F, Nanki T, Imai T, Kikuchi H, Hirohata S, Kohsaka H, et al. Inhibition of CX3CL1 (fractalkine) improves experimental autoimmune myositis in SJL/J mice. J Immunol. 2005;175:6987–96.
Place N, Ivarsson N, Venckunas T, Neyroud D, Brazaitis M, Cheng AJ, et al. Ryanodine receptor fragmentation and sarcoplasmic reticulum Ca2+ leak after one session of high-intensity interval exercise. Proc Natl Acad Sci U S A. 2015;112:15492–7.
Yamada T, Inashima S, Matsunaga S, Nara I, Kajihara H, Wada M. Different time course of changes in sarcoplasmic reticulum and myosin isoforms in rat soleus muscle at early stage of hyperthyroidism. Acta Physiol Scand. 2004;180(1):79–87.
Yamada T, Himori K, Tatebayashi D, Yamada R, Ashida Y, Imai T, et al. Electrical stimulation prevents preferential skeletal muscle myosin loss in steroid-denervation rats. Front Physiol. 2018;9:1111.
Allen DG, Kurihara S. The effects of muscle length on intracellular calcium transients in mammalian cardiac muscle. J Physiol. 1982;327:79–94.
Watanabe D, Wada M. Predominant cause of prolonged low-frequency force depression changes during recovery after in situ fatiguing stimulation of rat fast-twitch muscle. Am J Physiol Regul Integr Comp Physiol. 2016;311:R919–29.
Moisescu DG, Thieleczek R. Calcium and strontium concentration changes within skinned muscle preparations following a change in the external bathing solution. J Physiol. 1978;275:241–62.
Smith L. Spectrophotometric assay of cytochrome c oxidase. Methods Biochem Anal. 1955;2:427–34.
Srere PA. Citrate synthase. Methods Enzymol. 1969;13:3–11.
Yamada R, Himori K, Tatebayashi D, Ashida Y, Ikezaki K, Miyata H, et al. Preconditioning contractions prevent the delayed onset of myofibrillar dysfunction after damaging eccentric contractions. J Physiol. 2018;596:4427–42.
Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248–54.
Sabina RL, Swain JL, Olanow CW, Bradley WG, Fishbein WN, DiMauro S, et al. Myoadenylate deaminase deficiency. Functional and metabolic abnormalities associated with disruption of the purine nucleotide cycle. J Clin Invest. 1984;73:720–30.
Sewry CA, Clerk A, Heckmatt JZ, Vyse T, Dubowitz V, Strong PN. Dystrophin abnormalities in polymyositis and dermatomyositis. Neuromuscul Disord. 1991;1:333–9.
Kinder TB, Heier CR, Tully CB, Van der Muelen JH, Hoffman EP, Nagaraju K, et al. Muscle weakness in myositis: microRNA-mediated dystrophin reduction in a myositis mouse model and human muscle biopsies. Arthritis Rheumatol. 2020;72:1170–83.
Loell I, Helmers SB, Dastmalchi M, Alexanderson H, Munters LA, Nennesmo I, et al. Higher proportion of fast-twitch (type II) muscle fibres in idiopathic inflammatory myopathies - evident in chronic but not in untreated newly diagnosed patients. Clin Physiol Funct Imaging. 2011;31:18–25.
Win S, Than TA, Fernandez-Checa JC, Kaplowitz N. JNK interaction with Sab mediates ER stress induced inhibition of mitochondrial respiration and cell death. Cell Death Dis. 2014;5:e989.
Dong Y, Zhang M, Wang S, Liang B, Zhao Z, Liu C, et al. Activation of AMP-activated protein kinase inhibits oxidized LDL-triggered endoplasmic reticulum stress in vivo. Diabetes. 2010;59:1386–96.
Fishbein WN. Myoadenylate deaminase deficiency: inherited and acquired forms. Biochem Med. 1985;33:158–69.
Sabina RL, Fishbein WN, Pezeshkpour G, Clarke PR, Holmes EW. Molecular analysis of the myoadenylate deaminase deficiencies. Neurology. 1992;42:170–9.
Coley W, Rayavarapu S, Pandey GS, Sabina RL, Van der Meulen JH, Ampong B, et al. The molecular basis of skeletal muscle weakness in a mouse model of inflammatory myopathy. Arthritis Rheum. 2012;64:3750–9.
Fischer H, Esbjörnsson M, Sabina RL, Strömberg A, Peyrard-Janvid M, Norman B. AMP deaminase deficiency is associated with lower sprint cycling performance in healthy subjects. J Appl Physiol (1985). 2007;103:315–22.
Cheng J, Morisaki H, Sugimoto N, Dohi A, Shintani T, Kimura E, et al. Effect of isolated AMP deaminase deficiency on skeletal muscle function. Mol Genet Metab Rep. 2014;1:51–9.
This research is supported by grants from the Japan Society for the Promotion of Science (T.Y., JP21H03292, K.H., JP20J01754), Swedish Heart Lung Foundation (D.C.A., 20160741; 20180803), Harald and Greta Jeanssons stiftelse (D.C.A.), Swedish Society for Medical Research (D.C.A., SSMF; S16-0159), Svenska Läkaresällskapet (D.C.A., SLS-891461), and Swedish Research Council (H.W., 2018-02576).
The authors declare that they have no competing interests.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Yamada, T., Ashida, Y., Tamai, K. et al. Improved skeletal muscle fatigue resistance in experimental autoimmune myositis mice following high-intensity interval training. Arthritis Res Ther 24, 156 (2022). https://doi.org/10.1186/s13075-022-02846-2