Shin-Da Lee

Eicosapentaenoic Acid Protects against Palmitic Acid-Induced Endothelial Dysfunction via Activation of the AMPK/eNOS Pathway

Recent studies have shown that free fatty acids are associated with chronic inflammation, which may be involved in vascular injury. The intake of eicosapentaenoic acid (EPA) can decrease cardiovascular disease risks, but the protective mechanisms of EPA on endothelial cells remain unclear. In this study, primary human umbilical vein endothelial cells (HUVECs) treated with palmitic acid (PA) were used to explore the protective effects of EPA. The results revealed that EPA attenuated PA-induced cell death and activation of apoptosis-related proteins, such as caspase-3, p53 and Bax. Additionally, EPA reduced the PA-induced increase in the generation of reactive oxygen species, the activation of NADPH oxidase, and the upregulation of inducible nitric oxide synthase (iNOS). EPA also restored the PA-mediated reduction of endothelial nitric oxide synthase (eNOS) and AMP-activated protein kinase (AMPK) phosphorylation. Using AMPK siRNA and the specific inhibitor compound C, we found that EPA restored the PA-mediated inhibitions of eNOS and AKT activities via activation of AMPK. Furthermore, the NF-κB signals that are mediated by p38 mitogen-activated protein kinase (MAPK) were involved in protective effects of EPA. In summary, these results provide new insight into the possible molecular mechanisms by which EPA protects against atherogenesis via the AMPK/eNOS-related pathway.

Using Low-Intensity Pulsed Ultrasound to Improve Muscle Healing After Laceration Injury: An in vitro and in vivo Study

The purpose of this study was to determine whether low-intensity pulsed ultrasound (LIPUS) could enhance the regeneration of myofibers and shorten the healing time in injured muscle. NIH C2C12 cells, a well-known myoblastic cell line, are subclones derived from the mouse myoblast cell line established from normal adult C3H mouse leg muscle. The cells differentiate rapidly and produce extensive contracting myotubes expressing characteristic muscle proteins. We exposed C2C12 cells to LIPUS therapy using the EXOGEN 2000+ system ultrasound apparatus (Exogen Inc., Piscataway, NJ, USA) with a total treatment of 20 min every 24 h. At intervals of 2, 4, 6 and 8 days, cell growth was measured by the increase in cell number and western blot analysis of myogenin and actin. Forty mice (C57BL10J+/+) were divided into five groups of eight animals each and used in the published laceration injury model. The gastrocnemius muscle of the left leg was lacerated in all the animals. The control group (sham ultrasound) did not undergo LIPUS therapy. The ultrasound 7-, 14-, 21- and 28-day groups (only changing the number of days during which the ultrasound was applied to the injured muscle) were treated with LIPUS (20 min/day) for 7, 14, 21 and 28 consecutive days, respectively. All animals were sacrificed at 4 weeks after the injury. Evaluation methods included muscle regeneration and muscle contractile properties. LIPUS therapy produced a significantly higher proliferative rate and cell number at days 6 and 8 (p < 0.05). Densitometric evaluation revealed an increase in myogenin and actin proteins in cells treated with LIPUS in the 4-, 6- and 8-day groups. The regeneration of myofibers, fast-twitch and tetanus of LIPUS-treated muscles (21 and 28 days) was significantly greater relative to control muscles. There was no major strength difference between the normal non-injured muscle and the group treated with LIPUS for 28 days. In conclusion, this was the first experimental study to show that LIPUS therapy is able to enhance the regeneration of myofibers with better physiologic performance in injured mice muscles after laceration, especially prior to postoperative week 4. Findings of this study demonstrate a scientific basis for future clinical trials and establish an indication for LIPUS in enhancing muscle healing after laceration injury.

Topical cooling (icing) delays recovery from eccentric exercise-induced muscle damage.

Abstract Tseng, C-Y, Lee, J-P, Tsai, Y-S, Lee, S-D, Kao, C-L, Liu, T-C, Lai, C-S, Harris, MB, and Kuo, C-H. Topical Cooling (Icing) Delays Recovery From Eccentric Exercise–Induced Muscle Damage. J Strength Cond Res 27(5): 1354–1361, 2013—It is generally thought that topical cooling can interfere with blood perfusion and may have positive effects on recovery from a traumatic challenge. This study examined the influence of topical cooling on muscle damage markers and hemodynamic changes during recovery from eccentric exercise. Eleven male subjects (age 20.2 ± 0.3 years) performed 6 sets of elbow extension at 85% maximum voluntary load and randomly assigned to topical cooling or sham groups during recovery in a randomized crossover fashion. Cold packs were applied to exercised muscle for 15 minutes at 0, 3, 24, 48, and 72 hours after exercise. The exercise significantly elevated circulating creatine kinase-MB isoform (CK-MB) and myoglobin levels. Unexpectedly, greater elevations in circulating CK-MB and myoglobin above the control level were noted in the cooling trial during 48–72 hours of the post-exercise recovery period. Subjective fatigue feeling was greater at 72 hours after topical cooling compared with controls. Removal of the cold pack also led to a protracted rebound in muscle hemoglobin concentration compared with controls. Measures of interleukin (IL)-8, IL-10, IL-1&bgr;, and muscle strength during recovery were not influenced by cooling. A peak shift in IL-12p70 was noted during recovery with topical cooling. These data suggest that topical cooling, a commonly used clinical intervention, seems to not improve but rather delay recovery from eccentric exercise–induced muscle damage.

The Effects of Altitude Training on the AMPK-Related Glucose Transport Pathway in the Red Skeletal Muscle of Both Lean and Obese Zucker Rats

INTRODUCTION The skeletal muscle AMP-activated protein kinase (AMPK)-related glucose transport pathway is involved in glucose homeostasis. AIM In this study, we examined whether obese control Zucker rats had abnormal expression of proteins in the LKB1-AMPK-AS160-GLUT4 pathway in red gastrocnemius muscle compared to that in lean (normal) control Zucker rats. We also compared the chronic training effects of exercise, hypoxia, and altitude training on this pathway in lean and obese rats. METHODS At sea level, lean and obese rats were divided into 4 groups for 6 weeks training as follows: 1) control; 2) exercise (progressive daily swimming-exercise training with comparable exercise signals between the two groups); 3) hypoxia (8 hours of daily 14% O2 exposure); and 4) exercise plus hypoxia (also called altitude training). Seven animals were used for each group. RESULTS The obese rats in the control group had higher body weights, elevated fasting insulin and glucose levels, and higher baseline levels of muscle AMPK and AS160 phosphorylation compared with those of lean control rats. For obese Zucker rats in the exercise or hypoxia groups, the muscle AMPK phosphorylation level was significantly decreased compared with that of the control group. For obese Zucker rats in the altitude training group, the levels of AMPK, AS160 phosphorylation, fasting insulin, and fasting glucose were decreased concomitant with an approximate 50% increase in the muscle GLUT4 protein level compared with those of the control group. In lean rats, the altitude training efficiently lowered fasting glucose and insulin levels and increased muscle AMPK and AS160 phosphorylation as well as GLUT4 protein levels. CONCLUSION Our results provide evidence that long-term altitude training may be a potentially effective nonpharmacological strategy for treating and preventing insulin resistance based on its effects on the skeletal muscle AMPK-AS160-GLUT4 pathway.

Oral hydroxycitrate supplementation enhances glycogen synthesis in exercised human skeletal muscle.

Glycogen stored in skeletal muscle is the main fuel for endurance exercise. The present study examined the effects of oral hydroxycitrate (HCA) supplementation on post-meal glycogen synthesis in exercised human skeletal muscle. Eight healthy male volunteers (aged 22·0 (se 0·3) years) completed a 60-min cycling exercise at 70-75 % VO₂max and received HCA or placebo in a crossover design repeated after a 7 d washout period. They consumed 500 mg HCA or placebo with a high-carbohydrate meal (2 g carbohydrate/kg body weight, 80 % carbohydrate, 8 % fat, 12 % protein) for a 3-h post-exercise recovery. Muscle biopsy samples were obtained from vastus lateralis immediately and 3 h after the exercise. We found that HCA supplementation significantly lowered post-meal insulin response with similar glucose level compared to placebo. The rate of glycogen synthesis with the HCA meal was approximately onefold higher than that with the placebo meal. In contrast, GLUT4 protein level after HCA supplementation was significantly decreased below the placebo level, whereas expression of fatty acid translocase (FAT)/CD36 mRNA was significantly increased above the placebo level. Furthermore, HCA supplementation significantly increased energy reliance on fat oxidation, estimated by the gaseous exchange method. However, no differences were found in circulating NEFA and glycerol levels with the HCA meal compared with the placebo meal. The present study reports the first evidence that HCA supplementation enhanced glycogen synthesis rate in exercised human skeletal muscle and improved post-meal insulin sensitivity.

Effects of short-term detraining on measures of obesity and glucose tolerance in elite athletes

Abstract Athletes frequently adjust their training volume in line with their athletic competition schedule, onset of sport injury, and retirement. Whether maintenance of partial training activity during the detraining period can preserve optimal body composition and insulin sensitivity is currently unknown. Sixteen elite kayak athletes (mean [Vdot]O2max: 58.5 ml · kg−1· min−1, s = 1.77) were randomly assigned to a totally detrained group (age: 20.8 years, s = 0.7; body mass index: 23.74, s = 0.54) or partially detrained group (age: 21.8 years, s = 0.7; body mass index: 23.20, s = 1.02), whereby totally detrained participants terminated their training routine completely and the partially detrained participants preserved ∼50% of their previous training duration with equivalent intensity for one month. Body mass, waist circumference, oral glucose tolerance test, insulin, leptin, cortisol, and testosterone were measured during the trained state and after detraining. Waist circumferences for both the partially detrained and totally detrained groups were significantly elevated after detraining, with no group difference. However, body mass was reduced in both groups. Significant elevations in the area under the curve for insulin and fasted leptin with detraining were observed. These changes were greater in the totally detrained participants. In conclusion, the present results show that maintaining partial training activity cannot prevent an increase in waist circumference. During the detraining period, the magnitude of increase in plasma insulin and leptin concentrations was regulated in an activity-dependent manner.

Altered insulin-mediated and insulin-like growth factor-1-mediated vasorelaxation in aortas of obese Zucker rats

Objective:Insulin and insulin-like growth factor-1 (IGF-1) have vasorelaxant effects in vivo, which is dependent on nitric oxide (NO) production. The aim of this study was to investigate the vasorelaxant responses mediated by insulin and/or IGF-1 in aortas of obese Zucker rats.Methods:The thoracic aortas of eight lean and eight obese Zucker rats (6 months old) were isolated for vasorelaxation analysis. Insulin-induced and IGF-1-induced vasorelaxant responses were evaluated by the isometric tension of aortic rings in the organ bathes. The roles of phosphatidylinositol 3-kinase (PI3K) and nitric oxide synthase (NOS) in vasorelaxant responses were examined by treating selective inhibitors, such as wortmannin (an inhibitor of PI3K) and N ω-nitro-L-arginine methyl ester (L-NAME, a NOS inhibitor). In addition, the vascular responses to sodium nitroprusside (SNP), a direct vasodilator of vascular smooth muscle, were examined.Results:The insulin-induced vasorelaxation in aortas of obese rats was significantly decreased, whereas the IGF-1-induced vasorelaxation was significantly increased, compared with that in lean rats. After the pre-administration of wortmannin or L-NAME, the altered insulin-induced or IGF-1-induced vasorelaxation was abolished. There was no significant difference in the SNP-induced vasorelaxation between lean and obese rats.Conclusion:Our findings suggested that the decreased insulin-mediated vasorelaxation in obese rats appeared to be counteracted by the increased IGF-1-mediated vasorelaxation. Furthermore, the NO-dependent pathway was involved in the altered vasorelaxant responses. However, the SNP-induced vasorelaxation was not changed in obese rats.

Anxiety, depression and quality of life (QoL) in patients with chronic dizziness

Mood disorders and the severity of dizziness often interact with each other. However, the impact of age on anxiety and depression in dizzy patients has rarely been explored. The purpose of this study was to evaluate the correlation between the Dizziness Handicap Inventory (DHI) and the Hospital Anxiety and Depression Scale (HADS) in a group of patients with dizziness. The statistical relationships between age and each scale were studied. This cross-sectional study revealed a high correlation between DHI subgroups and HADS subscales. We found statistical correlation between neither age and HADS subscales nor age and DHI subgroups. In addition, Short-Form 36 Health Survey Questionnaires (SF-36) showed a high correlation with the dizziness handicap severity. The present study revealed that age does not influence mood disorders and the severity of dizziness in dizzy patients.

Effect of Mild Intermittent Hypoxia on Glucose Tolerance, Muscle Morphology and AMPK-PGC-1α Signaling

The main goal of this study was to investigate the long-term effect of daily 8-hour mild intermittent hypoxia (14-15% O2) on glucose tolerance and muscle morphology of Sprague-Dawley rats. The involvement of AMPK-PGC-1alpha-VEGF signaling pathways in the skeletal muscle was also determined during the first 8 hours of hypoxia. We found that mRNA levels of VEGF and PGC-1alpha were significantly increased above control after 8-h mild hypoxia without a change in AMPK phosphorylation. After 8 weeks of mild intermittent hypoxia treatment, plasma glucose and insulin levels in oral glucose tolerance test (OGTT), epididymal fat mass, and body weight were significantly lower compared to the control group. While soleus muscle weight was not changed, capillary and fiber densities in the hypoxia group were 33% and 35% above the control suggesting reorganization of muscle fibers. In conclusion, our data provide strong evidence that long-term mild intermittent hypoxia decreases the diffusion distance of glucose and insulin across muscle fibers, and decreases adiposity in rats. These changes may account for the improved glucose tolerance observed following the 8-week hypoxia treatment, and provides grounds for investigating the development of a mild non-pharmacological intervention in the treatment of obesity and type 2 diabetes.

Aerobic exercise acutely improves insulin‐ and insulin‐like growth factor‐1‐mediated vasorelaxation in hypertensive rats

Limited information is available concerning the effects of aerobic exercise on vasorelaxation in hypertension. The aim of this study was to investigate the effects of a single bout of aerobic exercise on insulin‐ and insulin‐like growth factor‐1 (IGF‐1)‐induced vasorelaxation in hypertensive rats. Four‐month‐old spontaneously hypertensive rats were randomly divided into a sedentary group (SHR) and an exercise group (SHR+Ex) subjected to a single bout of aerobic exercise conducted by treadmill running at 21 m min−1 for 1 h. Age‐matched Wistar–Kyoto rats were used as a normotensive control group (WKY). Insulin‐ and IGF‐1‐induced vasorelaxant responses in the three groups were evaluated by using isolated aortic rings, with or without endothelial denudation, in organ baths. Possible roles of phosphatidylinositol 3‐kinase (PI3K) and nitric oxide synthase (NOS) involved in the NO‐dependent vasorelaxation were examined by adding selective inhibitors. The role of superoxide was also clarified by adding superoxide dismutase (SOD). In addition, the endothelium‐independent vascular responses to sodium nitroprusside (SNP), a NO donor, were examined. The insulin‐ and IGF‐1‐induced vasorelaxation was significantly (P < 0.05) decreased in the SHR group compared with the WKY group. This decreased response in SHR was improved by exercise. These vasorelaxant responses among the three groups became similar after endothelial denudation and pretreatment with the PI3K inhibitor, NOS inhibitor or SOD. Also, no difference among groups was found in the SNP‐induced vasorelaxation. We concluded that a single bout of aerobic exercise acutely improves insulin‐ and IGF‐1‐mediated vasorelaxation in an endothelium‐dependent manner in hypertensive rats.