Enhanced γ-aminobutyric acid and sialic acid in fermented deer antler velvet and immune promoting effects.

Deer antler velvet is widely used in traditional medicine for its anti-aging, antioxidant, and immunity-enhancing effects. However, few studies have reported on the discovery of probiotic strains for deer antler fermentation to increase functional ingredient absorption. This study evaluated the ability of probiotic lactic acid bacteria to enhance the concentrations of bioactive molecules (e.g., sialic acid and gamma-aminobutyric acid [GABA]) in extracts of deer antler velvet. Seventeen strains of Lactobacillus spp. that were isolated from kimchi and infant feces, including L. sakei, L. rhamnosus, L. brevis, and L. plantarum, and those that improved the life span of Caenorhabditis elegans were selected for evaluation. Of the 17 strains, 2 (L. rhamnosus LFR20-004 and L. sakei LFR20-007) were selected based on data showing that these strains increased both the sialic acid and GABA contents of deer antler extract after fermentation for 2 d and significantly improved the life span of C. elegans. Co-fermentation with both strains further increased the concentrations of sialic acid, GABA, and metabolites such as short-chain fatty acids and amino acids. We evaluated the biological effects of the fermented antler velvet (FAV) on the antibacterial immune response in C. elegans by assessing worm survival after pathogen infection. The survival of the C. elegans conditioned with FAV for 24 h was significantly higher compared with that of the control worm group fed only normal feed (non-pathogenic E. coli OP50) exposed to E. coli O157:H7, Salmonella typhi, and Listeria monocytogenes. To evaluate the protective effects of FAV on immune response, cyclophosphamide (Cy), an immune-suppressing agent was treated to in vitro and in vivo. We found that FAV significantly restored viability of mice splenocytes and immune promoting-related cytokines (interleukin [IL]-6, IL-10, inducible nitric oxide synthase [iNOS], interferon [IFN]-γ, and tumor necrosis factor [TNF]-α) were activated compared to non-fermented deer antlers. This finding indicated the protective effect of FAV against Cy-induced cell death and immunosuppressed mice. Taken together, our study suggests that immune-promoting antler velvet can be produced through fermentation using L. rhamnosus LFR20-004 and L. sakei LFR20-007.

Effect of Porcine Whole Blood Protein Hydrolysate on Slow-Twitch Muscle Fiber Expression and Mitochondrial Biogenesis via the AMPK/SIRT1 Pathway.

Skeletal muscle is a heterogeneous tissue composed of a variety of functionally different fiber types. Slow-twitch type I muscle fibers are rich with mitochondria, and mitochondrial biogenesis promotes a shift towards more slow fibers. Leucine, a branched-chain amino acid (BCAA), regulates slow-twitch muscle fiber expression and mitochondrial function. The BCAA content is increased in porcine whole-blood protein hydrolysates (PWBPH) but the effect of PWBPH on muscle fiber type conversion is unknown. Supplementation with PWBPH (250 and 500 mg/kg for 5 weeks) increased time to exhaustion in the forced swimming test and the mass of the quadriceps femoris muscle but decreased the levels of blood markers of exercise-induced fatigue. PWBPH also promoted fast-twitch to slow-twitch muscle fiber conversion, elevated the levels of mitochondrial biogenesis markers (SIRT1, p-AMPK, PGC-1α, NRF1 and TFAM) and increased succinate dehydrogenase and malate dehydrogenase activities in ICR mice. Similarly, PWBPH induced markers of slow-twitch muscle fibers and mitochondrial biogenesis in C2C12 myotubes. Moreover, AMPK and SIRT1 inhibition blocked the PWBPH-induced muscle fiber type conversion in C2C12 myotubes. These results indicate that PWBPH enhances exercise performance by promoting slow-twitch muscle fiber expression and mitochondrial function via the AMPK/SIRT1 signaling pathway.

Betaine, a component of Lycium chinense, enhances muscular endurance of mice and myogenesis of myoblasts.

Sarcopenia is a disease characterized by the loss of muscle mass and function that occurs mainly in older adults. The present study was designed to investigate the hypothesis that water extract of Lycium chinense (WELC) would improve muscle function and promote myogenesis for sarcopenia. We investigated the effect of water extracts of L. chinense on muscular endurance function and myogenesis to examine its efficacy in sarcopenia. Intake of WELC-containing cheese enhanced the muscular endurance function of mice in treadmill endurance tests. In addition, the cross-sectional areas of muscle fibers in the gastrocnemius muscle of L. chinense-fed mice were greater than that of control mice. Furthermore, WELC and its key component marker substance betaine promoted myogenesis of myoblasts by increasing the expression of the myogenic protein myosin heavy chain 3 (Myh3) and myotube formation. Taken together, our results suggest that L. chinense may potentially be useful in the development of preventive and therapeutic agents for sarcopenia, as well as in providing basic knowledge on myogenesis and muscular functions.

Erratum to: Comparisons of Chemical Composition, Flavor and Bioactive Substances between Korean and Imported Velvet Antler Extracts.

[This corrects the article DOI: 10.5851/kosfa.2021.e4.].

Comparisons of Chemical Composition, Flavor and Bioactive Substances between Korean and Imported Velvet Antler Extracts.

The aim of this study was to compare the antioxidant activity, chemical composition, flavor and bioactive compounds between Korean and imported velvet antlers (VAs)-derived extracts. The Korean (KVA), Russian (RVA) and New Zealand (NZVA) VAs (n=24 each, dry form) purchased from a local supplier were used in the investigation. After extracting with water (750 g VA with 6,000 mL water) for 20 h at 95°C, the VA extracts (VAE) were then used for analysis of antioxidant activity, amino acids (AAs), flavor and bioactive compounds. Compared to the RVA and NZVA, the KVA extract showed significantly higher 2,2-diphenyl 1 picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid (ABTS) radicals scavenging activities (p<0.05). Significantly higher Fe content was found in the KVA while, higher Mn, Zn and Ca contents were found in the RVA (p<0.05). Twenty AAs were detected in all three VAEs and some of them (e.g., glycine and alanine) were higher in the KVA (p<0.05). A higher diversity (quality and quantity) of flavor compounds was found in the KVA extract compared to the imported VAs-derived extracts. Over six hundred metabolic compounds were identified in the VAEs. Among them, 412 compounds were commonly found in all the VAE types while, 109, 107, and 84 biomarker compounds were only found in the KVA, NZVA, and RVA extracts, respectively. Based on the results obtained in this study, it may be concluded that the country of origin partly affected the antioxidant activity, chemical composition, flavor and bioactive compounds of the VAEs.

Effects of Gouda cheese and Allium hookeri on thermogenesis in mice.

Cheese contains various beneficial nutrients, including calcium and whey protein, as well as large amounts of saturated fatty acids. Thus, intake of cheese increases the production of low-density lipoprotein-cholesterol (LDL-C), a well-defined risk factor for cardiovascular disease. Therefore, identification of natural products that inhibit LDL-C production following cheese intake and verification of the efficacy of such products in animal models are essential. Here, we evaluated the effects of Allium hookeri, a well-known traditional herbal remedy, on metabolism and thermogenesis in mice consuming a cheese-containing diet. Intake of A. hookeri extracts significantly blocked increases in body weight and fat mass caused by intake of Gouda cheese in mice. Additionally, increases in blood triglyceride levels following intake of Gouda cheese were alleviated by A. hookeri. Moreover, intake of Gouda cheese enhanced thermogenesis efficiency. Thus, A. hookeri may have applications as an important additive for reducing the risk of metabolic disease resulting from cheese consumption.

Biochemical activity of magnesium ions on human osteoblast migration.

Magnesium is well known as a biodegradable biomaterial that has been reported to promote bone remodeling in several studies; however, the underlying biological mechanism remains unclear. In the present study, the role of magnesium ions in the migration of U-2 OS cells, which are osteoblast-like cell lines, was investigated. Magnesium treatment did not significantly alter the global transcriptome of U-2 OS cells, but increased the protein expression level of SNAI2, an epithelial-mesenchymal transition (EMT) marker. In addition, it was confirmed that the junctional site localization of Zona-occludens 1 (ZO-1), a representative tight junction protein, was destroyed by magnesium treatment; furthermore, it was determined that cytoplasmic localization increased, and alkaline phosphatase (ALP) activity increased. The obtained results on the mechanism by which magnesium is involved in osteoblast migration, which is important for fracture healing, will contribute to the understanding of the bone-formation process in patients with osteoporosis and musculoskeletal injury.

Identification of Differentially Expressed Genes Associated with Extracellular Matrix Degradation and Inflammatory Regulation in Calcific Tendinopathy Using RNA Sequencing.

Calcific tendinopathy (CT), developed due to calcium hydroxyapatite deposition in the rotator cuff tendon, mostly affects women in their 40 s and 50 s and causes severe shoulder pain. However, the molecular basis of its pathogenesis and appropriate treatment methods are largely unknown. In this study, we identified 202 differentially expressed genes (DEGs) between calcific and adjacent normal tendon tissues of rotator cuff using RNA sequencing-based transcriptome analysis. The DEGs were highly enriched in extracellular matrix (ECM) degradation and inflammation-related processes. Further, matrix metalloproteinase 9 (MMP9) and matrix metalloproteinase 13 (MMP13), two of the enzymes associated with ECM degradation, were found to be highly upregulated 25.85- and 19.40-fold, respectively, in the calcific tendon tissues compared to the adjacent normal tendon tissues. Histopathological analyses indicated collagen degradation and macrophage infiltration at the sites of calcific deposit in the rotator cuff tendon. Our study acts as a foundation that may help in better understanding of the pathogenesis associated with CT, and thus in better management of the disease.

A combination of postmortem ageing and sous vide cooking following by blowtorching and oven roasting for improving the eating quality and acceptance of low quality grade Hanwoo striploin.

OBJECTIVE: It is well recognized that beef cuts from a low quality grade are usually associated with tougher, drier and less flavorful. Thus, the present study aimed at investigating the combined effects of postmortem ageing and sous vide (SV) cooking followed by oven roasting or blowtorching on the eating quality of low quality grade Hanwoo beef striploins. METHODS: Hanwoo beef striploins (quality grade 3) obtained from 36 month-old Hanwoo steers were used, and the samples were chiller aged for 0 and 14 d at 4°C. After ageing, the samples were prepared into 2.5-cm steaks which were then SV cooked at 55°C for 5 h and then raised to 60°C for 1 h, and thereafter the SV-cooked the steaks were further roasted in oven for 20 min (SV+OV) or blowtorched (SV+TC) for 2 min. The cooked samples were analyzed for microbiological quality, browning index, Wanrner-Bratzler shear force (WBSF), aroma flavor compounds and sensory properties. RESULTS: The SV cooking significantly reduced the WBSF values in beef samples (p<0.05). Blowtorching after SV cooking led to a browner surface of the beef steaks (p<0.05). The samples treated with SV+OV or SV+TC exhibited higher levels of Maillard reaction-derived aroma flavor compounds such as; pyrazines and sulfur-containing compounds compared to those just SV cooked. More especially, the SV+OV- or SV+TC- treated samples presented significantly higher flavor and overall acceptability scores compared to those just SV cooked (p<0.05). Ageing beef for 14 d significantly improved the tenderness by reducing the WBSF and increasing the tenderness scores. CONCLUSION: Thus, the combination of postmortem ageing and SV cooking followed by additional treatments (blowtorching or oven roasting) could be used to improve the eating quality especially tenderness and flavor as well as overall acceptability of low grade Hanwoo beef.

Effects of Traditional Sauce Type and Storage Time on Quality Characteristics, Shelf-life and Flavor Compounds of Marinated Pork Cooked by Sous Vide Method.

The present study aimed at evaluating effects of traditional sauce type and storage time on shelf-life and flavor compounds of marinated pork cooked by Sous-Vide method. Five different traditional sauces (Meju soy sauce, Brewed soy sauce, Fish-soy sauce, Ishiru fish sauce and Anchovy fish sauce) purchased from Asian countries were used. After marination with the sauces, polyvinylchloride film bags containing the marinated pork samples were cooked using Sous-Vide method for 55°C for 5 h and 60°C for 30 min, and were then stored for 8 wk at 10°C. Results showed that the pork samples marinated with the sauces retarded the growth of total plate counts (TPC) during storage. At 8th wk storage, TPC counts were significantly lower in all samples marinated with the sauces compared to control (p<0.05). Lipid oxidation level was significantly lower in the T2 (Meju soy sauce) and T6 (Anchovy fish sauce) compared to those of T3 (Brewed soy sauce) and T5 (Ishiru fish sauce) or control after 8 wk storage (p>0.05). Forty volatile flavor compounds were detected from the control and marinated samples at 4th wk of storage. The pork marinated with Anchovy fish sauce presented significantly higher amounts of importantly pleasant flavor compounds such as; pyrazines and sulfur-containing compounds than those marinated with other remaining sauces and control. It is concluded that the marination with Anchovy fish sauce partly improved the shelf-life and increased amounts of pleasant flavor compounds of Sous-Vide cooked pork products during storage in comparison to the other remaining sauces.

Platycodon grandiflorum-derived saponin enhances exercise function, skeletal muscle protein synthesis, and mitochondrial function.

Lower physical performance is an important risk factor in hypokinetic-related chronic disease, metabolic syndrome, and muscle atrophy. Our previous research demonstrated that Platycodon grandiflorum-derived saponin (PS) protects against eccentric exercise-induced muscle damage and mitochondrial function-related peroxisomal acyl-coenzme A oxidase (ACOX-1) and carnitine palmitoyltransferase (CPT-1) in high-fat diet-induced non-alcoholic steatohepatitis, and it inhibits osteoclast differentiation. However, the effects of PS on physical performance remain unknown. Therefore, we investigated whether PS enhances physical activity and skeletal muscle function. Supplementation with PS (2 mg/kg for 4 weeks) increased grip strength, wheel running repetition, and time to exhaustion in treadmill and swimming exercises. Marked increases in the synthesis of skeletal muscle proteins and muscle stem cell-related paired-box 7 (PAX7) were observed, and a decrease in the negative regulator myostatin was associated with enhanced muscle regeneration. Furthermore, PS induced expression of mitochondrial function proteins, including OXPHOS-III and -IV, in vivo and in vitro. These results suggest that PS enhances exercise function by ameliorating skeletal muscle protein synthesis and mitochondrial function.

Platycodon grandiflorum-derived saponin attenuates the eccentric exercise-induced muscle damage.

Platycodon grandiflorum contains triterpenoid saponins, such as platycodin D and platyconic acid A, and acts as a multifold nutritious compound. Our previous research demonstrated that Platycodon grandiflorum-derived saponin (PS) improves high-fat diet-induced non-alcoholic steatohepatitis and inhibits osteoclast differentiation. The pivotal effects of PS on inflammatory mechanism were suppressed NF-κB and matrix metalloproteinase (MMPs). However, the effects of PS on skeletal muscle damage remain unknown. Therefore, we investigated whether PS protects against eccentric exercise-induced muscle damage. A significant reduction in eccentric exercise-induced muscle damage area and muscle damage related to the level of NF-κB p65 by PS was associated with the downregulation of ERK/p38/SMAD signaling. Eccentric exercise caused muscle damage by increasing the serum lactate dehydrogenase (LDH), creatinine kinase (CK) and C-related protein level. The serum LDH, CK and C-related protein level was significantly lower in the PS supplementation group compared with the control group. Moreover, PS was inhibited MMP-1, MMP-2 and MMP-9. PS protects against eccentric exercise-induced muscle damage. Together, these results provide a novel perspective on the biological function of PS against muscle damage.

Leptin induces CREB-dependent aromatase activation through COX-2 expression in breast cancer cells.

Leptin plays a key role in the control of adipocyte formation, as well as in the associated regulation of energy intake and expenditure. The goal of this study was to determine if leptin-induced aromatase enhances estrogen production and induces tumor cell growth stimulation. To this end, breast cancer cells were incubated with leptin in the absence or presence of inhibitor pretreatment, and changes in aromatase and cyclooxygenase-2 (COX-2) expression were evaluated at the mRNA and protein levels. Transient transfection assays were performed to examine the aromatase and COX-2 gene promoter activities and immunoblot analysis was used to examine protein expression. Leptin induced aromatase expression, estradiol production, and promoter activity in breast cancer cells. Protein levels of phospho-STAT3, PKA, Akt, ERK, and JNK were increased by leptin. Leptin also significantly increased cAMP levels, cAMP response element (CRE) activation, and CREB phosphorylation. In addition, leptin induced COX-2 expression, promoter activity, and increased the production of prostaglandin E2. Finally, a COX-2 inhibitor and aromatase inhibitor suppressed leptin-induced cell proliferation in MCF-7 breast cancer cells. Together, our data show that leptin increased aromatase expression in breast cancer cells, which was correlated with COX-2 upregulation, mediated through CRE activation and cooperation among multiple signaling pathways.

Evaluation of Estrogenic Activity of Extract from the Herbal Mixture Cynanchum wilfordii Hemsley, Phlomis umbrosa Turczaninow, and Angelica gigas Nakai.

Hormone replacement therapy (HRT) consists of highly effective prescription medications for treating menopausal symptoms; however, these agents have exhibited side effects including the risk of estrogen-induced carcinogenesis. Therefore, interest in phytotherapy-based materials as a natural source of alternatives to estrogen therapy has increased. However, some of these herbal medicines have been reported to increase the risk of estrogen-induced cancer. Herbal formulations composed of a combination of Cynanchum wilfordii Hemsley (CW), Phlomis umbrosa Turczaninow (PU), and Angelica gigas Nakai (AG) extracts (CPAE) have been used for treating menopausal symptoms. Therefore, in this study, we aimed to examine the safety of CPAE by determining its potential adverse estrogenic activity using the Organization for Economic Cooperation and Development (OECD) test guideline 455 (TG455) in a stably transfected transcriptionally activated human estrogen receptor α (hERα)-HeLa9903 cell model. We found that CPAE did not how any estrogenic activity or stimulate promoters containing estrogen response elements in MCF-7 cells. In addition, CPAE showed no significant selective activity against hERα and hERß, non-selective activity against the ER, or effects on ER target gene expression. Furthermore, CPAE did not significantly induce MCF-7 cell proliferation and uterine weight increase in ovariectomized rats. These results demonstrate that CPAE can be used as beneficial herbal drug for prevention and therapeutic intervention of estrogen carcinogenesis in menopausal women.

Capsaicin Inhibits Dimethylnitrosamine-Induced Hepatic Fibrosis by Inhibiting the TGF-ß1/Smad Pathway via Peroxisome Proliferator-Activated Receptor Gamma Activation.

Capsaicin (CPS) exerts many pharmacological effects, but any possible influence on liver fibrosis remains unclear. Therefore, we evaluated the inhibitory effects of CPS on dimethylnitrosamine (DMN) and TGF-ß1-induced liver fibrosis in rats and hepatic stellate cells (HSCs). CPS inhibited DMN-induced hepatotoxicity, NF-κB activation, and collagen accumulation. CPS also suppressed the DMN-induced increases in α-SMA, collagen type I, MMP-2, and TNF-α. In addition, CPS inhibited DMN-induced TGF-ß1 expression (from 2.3 ± 0.1 to 1.0 ± 0.1) and Smad2/3 phosphorylation (from 1.5 ± 0.1 to 1.1 ± 0.1 and from 1.6 ± 0.1 to 1.1 ± 0.1, respectively) by activating Smad7 expression (from 0.1 ± 0.0 to 0.9 ± 0.1) via PPAR-γ induction (from 0.2 ± 0.0 to 0.8 ± 0.0) (p < 0.05). Furthermore, in HSCs, CPS inhibited the TGF-ß1-induced increases in α-SMA and collagen type I expression, via PPAR-γ activation. These results indicate that CPS can ameliorate hepatic fibrosis by inhibiting the TGF-ß1/Smad pathway via PPAR-γ activation.

Platycodin D Inhibits Osteoclastogenesis by Repressing the NFATc1 and MAPK Signaling Pathway.

Platycodon grandiflorum root-derived saponins (Changkil saponins, CKS) are reported to have many pharmacological activities. In our latest research, CKS was proven to have a significant osteogenic effect. However, the detail molecular mechanism of CKS on osteoclastic differentiation has not been fully investigated. Administration of CKS considerably reduced OVX-induced bone loss, and ameliorated the reduction in plasma levels of alkaline phosphatase, calcium, and phosphorus observed in OVX mice. CKS also repressed the deterioration of bone trabecular microarchitecture. Interestingly, platycodin D, the most abundant and major pharmacological constituent of triterpenoid CKS, inhibited receptor activator of NF-κB ligand (RANKL)-induced activation of NF-κB, and ERK and p38 MAPK, ultimately repressing osteoclast differentiation. OVX-induced bone turnover was attenuated by CKS, possibly via repression of osteoclast differentiation by platycodin D, the active component of CKS. Platycodin D can be regarded as an antiosteoporotic candidate for treatment of osteoporosis diseases. J. Cell. Biochem. 118: 860-868, 2017. © 2016 Wiley Periodicals, Inc.

Autophagic response to exercise training in skeletal muscle with age

Autophagy, a highly conserved quality control mechanism, is essential for the maintenance of cellular homeostasis and for the orchestration of an efficient cellular response to stress. During aging, the efficiency of autophagic degradation declines, and intracellular waste products accumulate. Therefore, in this study, we tested the hypothesis that skeletal muscle from old mice would have decreased autophagosome formation when compared to the muscle from young mice. We also examined whether autophagic regulatory events differ between muscle fiber types and in response to exercise in aged male mice. The extensor digitorum longus (EDL) and gastrocnemius muscles were studied in young and old ICR mice. Exercise was performed by allowing the mice to run on a treadmill with a 5° incline at 16.4 m/min for 40 min/day, 5 days/week for 8 weeks after a 1-week adaptation period. Our results indicated that the levels of microtubule-associated protein 1b light chain 3, a marker of autophagosome formation, were lower in both the EDL and the gastrocnemius muscle of old mice compared to those young mice. To identify the factors related to the changes observed, the expression of autophagy regulatory proteins was examined in the EDL and gastrocnemius muscles. Beclin-1, autophagy-related gene 7 (ATG7), and lysosome-associated membrane protein were found to be lower in the EDL and gastrocnemius muscles of old mice compared to those in the young mice, then Beclin-1, ATG7, and muscle-specific RING finger protein-1 upregulated after regular exercise. Moreover, the muscle weight/body weight was significantly increased only in the gastrocnemius muscle of the old trained mice. These data suggest that autophagy regulatory events are attenuated in old skeletal muscle. However, this effect is upregulated when animals are subjected to exercise training (AU)

Effect of treadmill exercise on interleukin-15 expression and glucose tolerance in zucker diabetic Fatty rats.

BACKGROUND: Interleukin-15 (IL-15), a well-known myokine, is highly expressed in skeletal muscle and is involved in muscle-fat crosstalk. Recently, a role of skeletal muscle-derived IL-15 in the improvement of glucose homeostasis and insulin sensitivity has been proposed. However, little is known regarding the influence of endurance training on IL-15 expression in type 2 diabetic skeletal muscles. We investigated the effect of endurance exercise training on glucose tolerance and IL-15 expression in skeletal muscles using type 2 diabetic animal models. METHODS: MALE ZUCKER DIABETIC FATTY (ZDF) AND ZDF LEAN CONTROL (ZLC) RATS WERE RANDOMLY DIVIDED INTO THREE GROUPS: sedentary ZLC, sedentary ZDF (ZDF-Con), and exercised ZDF (ZDF-Ex). The ZDF-Ex rats were forced to run a motor-driven treadmill for 60 minutes once a day 5 times per week for 12 weeks. Intraperitoneal glucose tolerance test (IPGTT) was performed after 12 weeks. Expression of IL-15 was measured using ELISA in extracted soleus (SOL) and gastrocnemius medial muscles. RESULTS: After 12 weeks of treadmill training, reduction of body weight was observed in ZDF-Ex compared to ZDF-Con rats. Glucose tolerance using IPGTT in diabetic rats was significantly improved in ZDF-Ex rats. Furthermore, the expression of IL-15 was significantly increased (P<0.01) only in the SOL of ZDF-Ex rats compared to ZDF-Con. Additionally, IL-15 expression in SOL muscles was negatively correlated with change of body weight (R=-0.424, P=0.04). CONCLUSION: The present study results suggest that 12 weeks of progressive endurance training significantly improved glucose tolerance with concomitant increase of IL-15 expression in SOL muscles of type 2 diabetic rats.

Effect of HX108-CS supplementation on exercise capacity and lactate accumulation after high-intensity exercise.

BACKGROUND: In the present study, we determined the effects of HX108-CS (mixed extract of Schisandra chinensis and Chaenomeles sinensis) supplementation on lactate accumulation and endurance capacity. Furthermore, we examined CK (creatine kinase), LDH (lactate dehydrogenase) activity to determine whether the HX108-CS affected markers of skeletal muscle injury in vivo and in vitro. METHODS: Exercise capacity was measured by an exhaustive swimming test using ICR mice divided into four groups; one group received distilled water (DW) (Control group, n = 10), and the other groups received three different dosages of HX108-CS (10, 50 and 100 mg/kg, n = 10 per group) solution in water orally. Then, for the time-dependent measurements of blood lactate, CK, and LDH, Sprague-Dawley rats were divided into two groups; one received DW (Control group, n = 10), and the other group received HX108-CS (100 mg/kg, n = 10) solution in the same way as mice. Before the exercise test, the animals were given either DW or HX108-CS for 2 weeks. High-intensity treadmill exercise was performed for 30 minutes. Blood samples were collected and analyzed during and after exercise. For the in vitro experiment, C2C12 cells were treated with HX108-CS to examine its effect on lactate production, CK, and LDH activity. RESULTS: Blood lactate concentration was significantly lowered immediately after treadmill exercise in HX108-CS group; however, there were no significant differences in activities of CK and LDH between HX108-CS and control during treadmill exercise and recovery phase. Furthermore, treatment with 100 mg/kg of HX108-CS led to a significant increase in the time to exhaustion in swimming test, and concurrently blood lactate concentration was significantly decreased in 50 and 100 mg/kg treated group. Moreover, our results of in vitro experiment showed that HX108-CS suppressed lactate production, CK, and LDH activity in a dose-dependent manner. CONCLUSIONS: These results suggest that supplementation with HX108-CS may enhance exercise capacity by lowering lactate accumulation. This may in part be related to an amelioration of skeletal muscle injury.

Autophagic response to exercise training in skeletal muscle with age.

Autophagy, a highly conserved quality control mechanism, is essential for the maintenance of cellular homeostasis and for the orchestration of an efficient cellular response to stress. During aging, the efficiency of autophagic degradation declines, and intracellular waste products accumulate. Therefore, in this study, we tested the hypothesis that skeletal muscle from old mice would have decreased autophagosome formation when compared to the muscle from young mice. We also examined whether autophagic regulatory events differ between muscle fiber types and in response to exercise in aged male mice. The extensor digitorum longus (EDL) and gastrocnemius muscles were studied in young and old ICR mice. Exercise was performed by allowing the mice to run on a treadmill with a 5° incline at 16.4 m/min for 40 min/day, 5 days/week for 8 weeks after a 1-week adaptation period. Our results indicated that the levels of microtubule-associated protein 1b light chain 3, a marker of autophagosome formation, were lower in both the EDL and the gastrocnemius muscle of old mice compared to those young mice. To identify the factors related to the changes observed, the expression of autophagy regulatory proteins was examined in the EDL and gastrocnemius muscles. Beclin-1, autophagy-related gene 7 (ATG7), and lysosome-associated membrane protein were found to be lower in the EDL and gastrocnemius muscles of old mice compared to those in the young mice, then Beclin-1, ATG7, and muscle-specific RING finger protein-1 upregulated after regular exercise. Moreover, the muscle weight/body weight was significantly increased only in the gastrocnemius muscle of the old trained mice. These data suggest that autophagy regulatory events are attenuated in old skeletal muscle. However, this effect is upregulated when animals are subjected to exercise training.