Baricitinib ameliorates inflammatory and neuropathic pain in collagen antibody-induced arthritis mice by modulating the IL-6/JAK/STAT3 pathway and CSF-1 expression in dorsal root ganglion neurons.

BACKGROUND: Janus kinase (JAK) inhibitors, such as baricitinib, are widely used to treat rheumatoid arthritis (RA). Clinical studies show that baricitinib is more effective at reducing pain than other similar drugs. Here, we aimed to elucidate the molecular mechanisms underlying the pain relief conferred by baricitinib, using a mouse model of arthritis. METHODS: We treated collagen antibody-induced arthritis (CAIA) model mice with baricitinib, celecoxib, or vehicle, and evaluated the severity of arthritis, histological findings of the spinal cord, and pain-related behaviours. We also conducted RNA sequencing (RNA-seq) to identify alterations in gene expression in the dorsal root ganglion (DRG) following baricitinib treatment. Finally, we conducted in vitro experiments to investigate the direct effects of baricitinib on neuronal cells. RESULTS: Both baricitinib and celecoxib significantly decreased CAIA and improved arthritis-dependent grip-strength deficit, while only baricitinib notably suppressed residual tactile allodynia as determined by the von Frey test. CAIA induction of inflammatory cytokines in ankle synovium, including interleukin (IL)-1ß and IL-6, was suppressed by treatment with either baricitinib or celecoxib. In contrast, RNA-seq analysis of the DRG revealed that baricitinib, but not celecoxib, restored gene expression alterations induced by CAIA to the control condition. Among many pathways changed by CAIA and baricitinib treatment, the interferon-alpha/gamma, JAK-signal transducer and activator of transcription 3 (STAT3), and nuclear factor kappa B (NF-κB) pathways were considerably decreased in the baricitinib group compared with the celecoxib group. Notably, only baricitinib decreased the expression of colony-stimulating factor 1 (CSF-1), a potent cytokine that causes neuropathic pain through activation of the microglia-astrocyte axis in the spinal cord. Accordingly, baricitinib prevented increases in microglia and astrocytes caused by CAIA. Baricitinib also suppressed JAK/STAT3 pathway activity and Csf1 expression in cultured neuronal cells. CONCLUSIONS: Our findings demonstrate the effects baricitinib has on the DRG in relation to ameliorating both inflammatory and neuropathic pain.

Gait disorders induced by photothrombotic cerebellar stroke in mice.

Patients with cerebellar stroke display relatively mild ataxic gaits. These motor deficits often improve dramatically; however, the neural mechanisms of this improvement have yet to be elucidated. Previous studies in mouse models of gait ataxia, such as ho15J mice and cbln1-null mice, have shown that they have a dysfunction of parallel fiber-Purkinje cell synapses in the cerebellum. However, the effects of cerebellar stroke on the locomotor kinematics of wild-type mice are currently unknown. Here, we performed a kinematic analysis of gait ataxia caused by a photothrombotic stroke in the medial, vermal, and intermediate regions of the cerebellum of wild-type mice. We used the data and observations from this analysis to develop a model that will allow locomotive prognosis and indicate potential treatment regimens following a cerebellar stroke. Our analysis showed that mice performed poorly in a ladder rung test after a stroke. During walking on a treadmill, the mice with induced cerebellar stroke had an increased duty ratio of the hindlimb caused by shortened duration of the swing phase. Overall, our findings suggest that photothrombotic cerebellar infarction and kinematic gait analyses will provide a useful model for quantification of different types of acute management of cerebellar stroke in rodents.

Greater arterial wall viscosity in endurance-trained men.

PURPOSE: The age-associated increase in arterial wall viscosity (AWV) is attenuated by high cardiorespiratory fitness level. However, AWV in endurance-trained athletes have not been determined. We designed a cross sectional study to compare central AWV and compliance between endurance-trained young athletes and age-matched control men. METHODS: Twenty-one endurance-trained men (age 20.7 ± 0.3 years) and 20 age-matched healthy control men (age 21.6 ± 0.4 years) were studied. The common carotid artery was measured noninvasively by tonometry and automatic tracking of B-mode images to obtain instantaneous pressure and diameter hysteresis loops, and we calculated the dynamic carotid arterial compliance, static (effective and isobaric) compliance, and viscosity index. RESULTS: The AWV index in the endurance-trained men was larger than the control peers (2285 ± 181 vs. 1429 ± 124 mmHg·s/mm: P < 0.001). In addition, dynamic and static compliance were not statistically different between both groups. CONCLUSION: The present study indicated that the central AWV in endurance-trained athletes was greater than age-matched healthy control men. We believe that the AWV, as well as arterial compliance, is an important element for assessing vascular adaptation to endurance training.

Alteration of gait parameters in a mouse model of surgically induced knee osteoarthritis.

PURPOSE: Joint pain is the most common symptom of osteoarthritis (OA); however, its mechanism remains unclarified. The present study investigated hindlimb motion during locomotion on the treadmill using a three-dimensional (3D) motion analysis system with high-speed cameras to evaluate whether this method can be used as an indication of joint pain in a mouse model of surgically induced OA. METHODS: We resected the medial meniscus and medial collateral ligament in 8-week old C57BL/6 male mice and performed locomotion recording 6 months post-operatively. Additionally, we performed the same recording after oral administration of the selective cyclooxygenase-2 inhibitor to determine whether alteration of the parameters were associated with joint pain. RESULTS: OA development, characterized by cartilage degeneration and osteophyte formation, was markedly enhanced in the OA group. There was no significant difference between the sham and OA groups in basic gait parameters, including stance duration, swing duration and gait cycle. However, when we divided the gait cycle into four phases and calculated the joint ranges of motion in each phase, the range of motion of the knee joint during the stepping-in phase and the swing duration were significantly decreased in the OA group. These significant differences between the sham and OA groups were diminished by the oral administration of a selective cyclooxygenase-2 inhibitor to the OA group. CONCLUSION: The present method may be useful to evaluate joint pain in experimental mice and contribute to elucidating the molecular mechanisms of pain in the OA knee joint in combination with genetically modified mice.

Relative contribution of organs other than brain to resting energy expenditure is consistent among male power athletes.

We have previously shown that resting energy expenditure (REE) adjusted by fat-free mass (FFM) in male college athletes remains consistent regardless of FFM. The FFM comprises internal organs with high metabolic activity, such as liver and brain, which account for 60 to 80% of REE in adults. The purpose of the present study is to examine the contribution of internal organs to the REE of the FFM fraction among male power athletes. The study included 37 American male college football players. REE was measured by indirect calorimetry and body composition was measured by dual energy X-ray absorptiometry (DXA). Mass of brain, liver, and kidneys was measured by MRI and mass of heart was estimated by echocardiography. Normal levels of thyroid hormone (triiodothyronine: T3) were confirmed in all subjects prior to the analysis. Multiple regression analysis was used to assess the influence of FFM, fat mass (FM), T3, and mass of organs on variance of REE. Average body weight and FFM were 81.2±11.3 kg and 67.7±7.4 kg, respectively. The relative contributions of liver, kidneys, and heart to REE were consistent regardless of FFM, while the REE of brain was negatively correlated with FFM (r=-0.672, p<0.001). Only FFM and T3 were found to be independent factors influencing REE. These results suggest that a steady contribution of internal organs other than the brain is the major reason for the consistency of the REE/FFM ratio in male power athletes.

Effects of resveratrol and SIRT1 on PGC-1α activity and mitochondrial biogenesis: a reevaluation.

It has been reported that feeding mice resveratrol activates AMPK and SIRT1 in skeletal muscle leading to deacetylation and activation of PGC-1α, increased mitochondrial biogenesis, and improved running endurance. This study was done to further evaluate the effects of resveratrol, SIRT1, and PGC-1α deacetylation on mitochondrial biogenesis in muscle. Feeding rats or mice a diet containing 4 g resveratrol/kg diet had no effect on mitochondrial protein levels in muscle. High concentrations of resveratrol lowered ATP concentration and activated AMPK in C2C12 myotubes, resulting in an increase in mitochondrial proteins. Knockdown of SIRT1, or suppression of SIRT1 activity with a dominant-negative (DN) SIRT1 construct, increased PGC-1α acetylation, PGC-1α coactivator activity, and mitochondrial proteins in C2C12 cells. Expression of a DN SIRT1 in rat triceps muscle also induced an increase in mitochondrial proteins. Overexpression of SIRT1 decreased PGC-1α acetylation, PGC-1α coactivator activity, and mitochondrial proteins in C2C12 myotubes. Overexpression of SIRT1 also resulted in a decrease in mitochondrial proteins in rat triceps muscle. We conclude that, contrary to some previous reports, the mechanism by which SIRT1 regulates mitochondrial biogenesis is by inhibiting PGC-1α coactivator activity, resulting in a decrease in mitochondria. We also conclude that feeding rodents resveratrol has no effect on mitochondrial biogenesis in muscle.

Organ size increases with weight gain in power-trained athletes.

The purpose of this study was to determine whether overfeeding and high-intensity physical training increase organ mass. We examined this question using cross-sectional and longitudinal studies in which we measured collegiate male American football players. Freshman (n = 10) and senior players in their second and third years of college (n = 17) participated in the cross-sectional study. The same measurements of the same freshman players (n = 10) were assessed after the one-year weight gain period in the longitudinal study. Fat-free mass (FFM), skeletal muscle, and adipose tissue mass were obtained using dual-energy X-ray absorptiometry. Liver, kidney, brain, and heart volumes were calculated using magnetic resonance imaging or echocardiography. Compared with the freshman players, the senior players had 10.8 kg more FFM, and 0.29 kg, 0.08 kg, and 0.09 kg greater liver, heart, and kidney mass, respectively. In the longitudinal study, FFM, liver, heart, and kidney mass of the freshman players increased by 5.2 kg, 0.2 kg, 0.04 kg, and 0.04 kg, respectively, after one year of overfeeding and physical training. On the other hand, the organ-tissue mass to FFM ratio did not change, except for the brain, in either the cross-sectional or longitudinal studies. Our results indicated that the organ-tissue masses increased with overfeeding and physical training in male collegiate American football players.

Effects of different modes of exercise on appetite and appetite-regulating hormones.

The present study determined the changes in appetite and appetite-regulating gut hormones during and following bouts of both rope skipping exercise (weight-bearing) and bicycle ergometer exercise (non-weight-bearing). After a 12-h fast, 15 young men (mean ± SD, age 24.4 ± 1.7 yrs, maximal oxygen uptake 47.0 ± 6.5 mL/kg/min) participated in three 160 min trials: (1) rope skipping exercise (295 ± 40 kcal, 3 sets × 10 min with 5-min interval, then rested for 120 min); (2) bicycle ergometer exercise (288 ± 36 kcal, 3 sets × 10 min with 5-min interval, then rested for 120 min); (3) control (rested for 160 min). Ratings of perceived hunger and acylated ghrelin were suppressed and total peptide YY (PYY) were increased during and immediately after exercise in both exercise trials, but glucagon liked peptide-1 was not changed. Furthermore, suppressed hunger during rope skipping exercise was greater than that during bicycle ergometer exercise, but there were no differences in acylated ghrelin and total PYY. These results indicate that weight-bearing exercise has a greater exercise-induced appetite suppressive effect compared with non-weight-bearing exercise, and both forms of exercise lowered acylated ghrelin and increased total PYY, but the changes did not differ significantly between exercise modes.

ß-Adrenergic stimulation does not activate p38 MAP kinase or induce PGC-1α in skeletal muscle.

There are reports that the ß-adrenergic agonist clenbuterol induces a large increase in peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) in skeletal muscle. This has led to the hypothesis that the increases in PGC-1α and mitochondrial biogenesis induced in muscle by endurance exercise are mediated by catecholamines. In the present study, we evaluated this possibility and found that injecting rats with clenbuterol or norepinephrine induced large increases in PGC-1α and mitochondrial proteins in brown adipose tissue but had no effect on PGC-1α expression or mitochondrial biogenesis in skeletal muscle. In brown adipocytes, the increase in PGC-1α expression induced by ß-adrenergic stimulation is mediated by activation of p38 mitogen-activated protein kinase (p38 MAPK), which phosphorylates and activates the cAMP response element binding protein (CREB) family member activating transcription factor 2 (ATF2), which binds to a cyclic AMP response element (CRE) in the PGC-1α promoter and mediates the increase in PGC-1α transcription. Phospho-CREB does not have this effect. Our results show that the reason for the lack of effect of ß-adrenergic stimulation on PGC-1α expression in muscle is that catecholamines do not activate p38 or increase ATF2 phosphorylation in muscle.

Habitual rowing exercise is associated with high physical fitness without affecting arterial stiffness in older men.

The present study elucidated the effects of habitual rowing exercise on arterial stiffness and plasma levels of the vasoconstrictor endothelin-1 and the vasodilator nitric oxide (NO) in older men. Eleven rowers (68.0 ± 1.6 years) and 11 sedentary control older men (64.9 ± 1.1 years) were studied. Peak oxygen uptake (36.0 ± 1.7 vs. 27.7 ±1.9 ml · kg(-1) · min(-1)), leg press power (1346 ± 99 vs. 1077 ± 68 W), and HDL-cholesterol (75 ± 5 vs. 58 ±3 mg · ml(-1)) were higher and triglyceride (78 ± 9 vs. 120 ± 14 mg · ml(-1)) was lower in rowers than in control participants (all P < 0.05). Arterial stiffness indices (carotid ß-stiffness and cardio-ankle vascular index) and plasma endothelin-1 and NOx (nitrite + nitrate) levels did not differ between the two groups. These results suggest that habitual rowing exercise in older men is associated with high muscle power and aerobic capacity, and favourable blood lipid profile without affecting arterial stiffness or plasma levels of endotheline-1 and NO.

Association between muscular strength and metabolic risk in Japanese women, but not in men.

We examined whether cardiorespiratory fitness (maximal oxygen uptake, VO(2)max) and muscular strength (grip strength) are associated with individual and clustered metabolic risk factors independently of abdominal adiposity in Japanese men (n=110) and women (n=110) aged 20-69 years. Blood pressure, triglycerides (TG), HDL cholesterol, and fasting plasma glucose (FPG) were assessed and metabolic risk score was calculated, which is the sum of the z scores for each individual risk factor. Waist circumference was measured and the area of visceral fat was assessed by MRI. Multiple linear regression analysis revealed that VO(2)max was inversely associated with TG in men (p<0.05) and grip strength was negatively associated with FPG and metabolic risk score in women (p<0.001 and p<0.05, respectively), independently of waist circumference. Adjusting for visceral fat instead of waist circumference, similar results were obtained in women (p<0.01 and p<0.05, respectively), but the association between VO(2)max and TG in men was attenuated to nonsignificant. This cross-sectional study demonstrates that muscular strength is inversely associated with plasma glucose levels and clustered metabolic risk factors independently of abdominal adiposity in Japanese women, but not in men.

Fat-free mass can be utilized to assess resting energy expenditure for male athletes of different body size.

The fat-free mass (FFM) of athletes is typically large, and thus the FFM is often utilized to estimate their resting energy expenditure (REE). While the proportional contribution of organ-tissues to the total influence of FFM on REE is known for untrained individuals and female athletes, the extent to which this is valid for male athletes is unclear. The purpose of this study was to clarify the contribution of the components of FFM to REE in male athletes. Fifty-seven male athletes participated in this study. REE was assessed by indirect calorimetry and body composition by dual X-ray absorptiometry. The athletes were equally divided into three groups based on FFM: Small (S), Medium (M), and Large (L). When measured REE (REEm) was compared with REE estimated (REEe) based on the four organ-tissue compartments with set metabolic rates, REEm and REEe had a strong association (r=0.76, p<0.001). In addition, the absolute value of total REE became larger in accordance with body size (S: 1,643±144, M: 1,865±140, and L: 2,060±156 kcal/d) accompanied by increases in mass of all four organ-tissue compartments as body size increased. The consistency of REE/FFM in male athletes in spite of the difference in body size can be explained by the steadiness among the three groups of the relative contribution of each organ-tissue compartment to the FFM. Based on these results, the FFM is the major determinant of REE regardless of body size in male athletes.

Visceral fat is a strong predictor of insulin resistance regardless of cardiorespiratory fitness in non-diabetic people.

Abdominal adiposity and low cardiorespiratory fitness are associated with insulin resistance in people with impaired glucose tolerance and type 2 diabetes. However, little is known about which factor precedes insulin resistance in people with impaired glucose tolerance and type 2 diabetes, and which is the stronger predictor of insulin resistance in non-diabetic people. The purpose of this study was to examine the relationship between insulin resistance and cardiorespiratory fitness, visceral fat, and subcutaneous fat in non-diabetic people. Subjects included 87 men and 77 women aged 30-72 y (mean+/-SD, 51.3+/-12.3 y). Cardiorespiratory fitness was assessed by measuring the maximal oxygen uptake (VO2max) in a progressive continuous test to exhaustion on a cycle ergometer. The visceral and subcutaneous fat areas were measured by magnetic resonance imaging. The homeostasis model assessment of insulin resistance (HOMA-R) was calculated from the fasting concentrations of glucose and insulin. Stepwise multiple linear regression analysis revealed that visceral and subcutaneous fat were significant correlates of HOMA-R, explaining 24% and 6% of the variance, respectively, whereas sex, age, and VO2max were not significant independent determinants. Abdominal fat deposition rather than cardiorespiratory fitness is a significant predictor of insulin resistance in non-diabetic people; visceral fat is the most important factor.

Elderly oarsmen have larger trunk and thigh muscles and greater strength than age-matched untrained men.

To evaluate whether regularly performed rowing exercise affects the trunk muscles size and function, and to examine the effect of rowing exercise on thigh muscle size and function in elderly rowers, we compared the cross-sectional area (CSA) and strength of these muscles in elderly male rowers and in age-matched untrained men. Participants were 16 elderly rowing-trained men (ROW age, 67.8 +/- 2.3 years) and 18 elderly untrained men (CON 66.2 +/- 3.0 years). CSA was measured by MRI in the trunk and thigh muscles. Isometric trunk flexion force and leg extension power were measured. ROW had a 20% larger total trunk muscle CSA than CON (P < 0.01); rectus abdominis was 27% larger, psoas major 64% larger, and erector spinae 14% larger in ROW than in CON (P < 0.05-0.001). Isometric trunk flexion force was related to the CSA of the rectus abdominis (r = 0.777, P < 0.001) and psoas major (r = 0.694, P < 0.001), and was 42% larger in ROW than in CON (P < 0.001). However, force adjusted for the CSA of the muscles did not differ significantly between CON and ROW. In ROW, the CSA was 13% larger in the total thigh muscles (P < 0.01), and leg extension power was 43% higher than in CON (P < 0.001). These results suggest that rowing exercise is a favorable training modality for the trunk muscles, especially psoas major and that it improves thigh muscle size and function in elderly men.