• Title/Summary/Keyword: myosin heavy chain

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Research article Black ginseng activates Akt signaling, thereby enhancing myoblast differentiation and myotube growth

  • Lee, Soo-Yeon;Go, Ga-Yeon;Vuong, Tuan Anh;Kim, Jee Won;Lee, Sullim;Jo, Ayoung;An, Jun Min;Kim, Su-Nam;Seo, Dong-Wan;Kim, Jin-Seok;Kim, Yong Kee;Kang, Jong-Sun;Lee, Sang-Jin;Bae, Gyu-Un
    • Journal of Ginseng Research
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    • v.42 no.1
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    • pp.116-121
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    • 2018
  • Background: Black ginseng (BG) has greatly enhanced pharmacological activities relative to white or red ginseng. However, the effect and molecular mechanism of BG on muscle growth has not yet been examined. In this study, we investigated whether BG could regulate myoblast differentiation and myotube hypertrophy. Methods: BG-treated C2C12 myoblasts were differentiated, followed by immunoblotting for myogenic regulators, immunostaining for a muscle marker, myosin heavy chain or immunoprecipitation analysis for myogenic transcription factors. Results: BG treatment of C2C12 cells resulted in the activation of Akt, thereby enhancing hetero-dimerization of MyoD and E proteins, which in turn promoted muscle-specific gene expression and myoblast differentiation. BG-treated myoblasts formed larger multinucleated myotubes with increased diameter and thickness, accompanied by enhanced Akt/mTOR/p70S6K activation. Furthermore, the BG treatment of human rhabdomyosarcoma cells restored myogenic differentiation. Conclusion: BG enhances myoblast differentiation and myotube hypertrophy by activating Akt/mTOR/p70S6k axis. Thus, our study demonstrates that BG has promising potential to treat or prevent muscle loss related to aging or other pathological conditions, such as diabetes.

Cryoprotective Effect and Mechanism of Corn Starch Enzyme Hydrolysates on Fish Protein 1. Cryoprotective Effect of Corn Starch Enzyme Hydrolysates on Fish Protein (전분가수분해물의 어육단백질 동결변성 방지효과 및 작용기구 1. 옥수수전분가수분해물의 어육단백질에 대한 동결변성 방지효과)

  • LEE Kang-Ho;JUNG Byung-Chun;HONG Byung-Il
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.6
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    • pp.823-828
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    • 1998
  • The objective of this study is to investigate cryoprotective effects of corn starch enzyme hydrolysates of nonsweet and low-calories on denaturation of frozen fish protein. The cryoprotective effects of were examined in Alaska pollack actomyosin solution by changes in SDS-PAGE pattern, solubility, and $Ca^{2+}$-ATPase activity. When samples stored for 0 and 30 days were compared on SDS-PAGE patterns, severe changes in all bands were shown on the control sample regardless of storage temperature, especially in myosin heavy chain (MHC). Not much difference no appeared the electrophoretic pattern in case of the samples containing sucrose at any storage temperature during 30 days of storage. The cryoprotective effect of the hydrolysates were markedly dependant on storage temperature and no MHC band was found in the samples stored at $-5^{\circ}C$. The SDS-PAGE patterns of sample stored at $-20^{\circ}C$, however, completely maintained after 30 days or storage. When the samples were stored at $-5^{\circ}C$, the solubility of the sample containing sucrose was retained at $90\%$ after 30 days of storage, whereas dramatically decreased in other samples. The samples including sucrose, D.E. 10, 15, and 20 revealed $90\%$ in solubility when stored at $-20^{\circ}C$. The tendency of remaining $Ca^{2+}$-ATPase activity was almost shown the same as that of solubility.

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Rheological Properties of Pork Myofibrillar Protein and Sodium Caseinate Mixture as Affected by Transglutaminase with Various Incubation Temperatures and Times (Transglutaminase를 첨가한 돈육 근원섬유단백질과 카제인염 혼합물의 배양온도와 시간에 따른 물성변화)

  • Hwang, Ji-Suk;Lee, Hong-Chul;Chin, Koo-Bok
    • Food Science of Animal Resources
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    • v.28 no.2
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    • pp.154-159
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    • 2008
  • To investigate the rheological properties of protein mixed gels mediated by microbial transglutaminase (MTGase), pork myofibrillar protein (MFP), sodium caseinate (SC) and their mixture (MS), the various gels were incubated at different temperatures for various times. Extracted MFP, SC and their mixture (MS, 1:1) were incubated at different temperatures ($4^{\circ}C$ vs $37^{\circ}C$) for various times (0, 0.5, 2, 4 hr), and assessed for viscosity, gel strength and other characteristics using differential scanning calorimeter (DSC) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). DSC measurements showed that incubation at $37^{\circ}C$ rather than $4^{\circ}C$ caused marked changes in thermal transition, and MS displayed similar thermal curves (three endothermic transitions) to MFP and SC alone. After incubation at $37^{\circ}C$ for 2 hrs, the viscosity (cP) of MS increased (p<0.05) due to induction by MTGase, whereas no differences were observed at $4^{\circ}C$. However, gel strength values were no different, regardless of incubation temperatures and times. Future research will address how longer incubation times affect the functionality of protein mixed gels mediated by MTGase.

Role of LPS-activated Macrophages in the Differentiation of Mesenchymal Stem Cells into Smooth Muscle Cells (중간엽 줄기세포의 평활근 세포로의 분화에서 LPS에 의해 활성화된 대식세포의 역할)

  • Lee, Mi Jeong;Do, Eun Kyoung;Kim, Jae Ho
    • Journal of Life Science
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    • v.23 no.1
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    • pp.137-142
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    • 2013
  • Human adipose-derived mesenchymal stem cells (hMSCs) are highly useful for vascular regeneration of injured or inflamed tissue. Lipopolysaccharide (LPS) is a potent activator of macrophages and stimulates macrophages to release inflammatory cytokines. In the present study, we explored the role of LPS-activated macrophages in the differentiation of hMSCs to smooth muscle cells (SMCs). We demonstrated that conditioned medium from LPS-induced macrophages (LPS CM) stimulates differentiation of hMSCs to SMCs, as evidenced by increased expression of smooth muscle-specific markers, including alpha-smooth muscle actin (${\alpha}$-SMA), smooth muscle-myosin heavy chain, and calponin. LPS induced the secretion of $PGF2{\alpha}$ from macrophages, and $PGF2{\alpha}$ treatment stimulated expression levels of SMC-specific markers in hMSCs. Furthermore, small interfering RNA-mediated silencing of the $PGF2{\alpha}$ receptor inhibited LPS CM-stimulated ${\alpha}$-SMA expression. These results suggest that LPS-activated macrophages promote differentiation of hMSCs to SMCs through a $PGF2{\alpha}$-dependent mechanism.

Comparison of Gene Expression Levels of Porcine Satellite Cells from Postnatal Muscle Tissue during Differentiation

  • Jeong, Jin Young;Kim, Jang Mi;Rajesh, Ramanna Valmiki;Suresh, Sekar;Jang, Gul Won;Lee, Kyung-Tai;Kim, Tae Hun;Park, Mina;Jeong, Hak Jae;Kim, Kyung Woon;Cho, Yong Min;Lee, Hyun-Jeong
    • Reproductive and Developmental Biology
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    • v.37 no.4
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    • pp.219-224
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    • 2013
  • Muscular satellite cell (SC), which is stem cell of postnatal pig, is an important for study of differentiation into adipogenesis, myogenesis, and osteoblastogenesis. In this study, we isolated and examined from pig muscle tissue to determine capacity in proliferate, differentiate, and expression of various genes. Porcine satellite cells (PSC) were isolated from semimembranosus (SM) muscles of 90~100 days old pigs according to standard conditions. The cell proliferation increased in multi-potent cell by Masson's, oil red O, and Alizarin red staining respectively. We performed the expression levels of differentiation related genes using real-time PCR. We found that the differentiation into adipocyte increased expression levels of both fatty acid binding protein 4 (FABP4) and peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) genes (p<0.01). Myocyte increased the expression levels of the myosin heavy chain (MHC), myogenic factor 5 (Myf5), myogenic regulatory factor (MyoD), and Myogenic factor 4 (myogenin) (p<0.01). Osteoblast increased the expression levels of alkaline phosphatase (ALP) (p<0.01). Finally, porcine satellite cells were induced to differentiate towards adipogenic, myogenic, and osteoblastogenic lineages. Our results suggest that muscle satellite cell in porcine may influence cell fate. Understanding the progression of PSC may lead to improved strategies for augmenting meat quality.

Isolation and identification of goose skeletal muscle satellite cells and preliminary study on the function of C1q and tumor necrosis factor-related protein 3 gene

  • Wang, Han;He, Ke;Zeng, Xuehua;Zhou, Xiaolong;Yan, Feifei;Yang, Songbai;Zhao, Ayong
    • Animal Bioscience
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    • v.34 no.6
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    • pp.1078-1087
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    • 2021
  • Objective: Skeletal muscle satellite cells (SMSCs) are significant for the growth, regeneration, and maintenance of skeletal muscle after birth. However, currently, few studies have been performed on the isolation, culture and inducing differentiation of goose muscle satellite cells. Previous studies have shown that C1q and tumor necrosis factor-related protein 3 (CTRP3) participated in the process of muscle growth and development, but its role in the goose skeletal muscle development is not yet clear. This study aimed to isolate, culture, and identify the goose SMSCs in vitro. Additionally, to explore the function of CTRP3 in goose SMSCs. Methods: Goose SMSCs were isolated using 0.25% trypsin from leg muscle (LM) of 15 to 20 day fertilized goose eggs. Cell differentiation was induced by transferring the cells to differentiation medium with 2% horse serum and 1% penicillin streptomycin. Immunofluorescence staining of Desmin and Pax7 was used to identify goose SMSCs. Quantitative realtime polymerase chain reaction and western blot were applied to explore developmental expression profile of CTRP3 in LM and the regulation of CTRP3 on myosin heavy chains (MyHC), myogenin (MyoG) expression and Notch signaling pathway related genes expression. Results: The goose SMSCs were successfully isolated and cultured. The expression of Pax7 and Desmin were observed in the isolated cells. The expression of CTRP3 decreased significantly during leg muscle development. Overexpression of CTRP3 could enhance the expression of two myogenic differentiation marker genes, MyHC and MyoG. But knockdown of CTRP3 suppressed their expression. Furthermore, CTRP3 could repress the mRNA level of Notch signaling pathway-related genes, notch receptor 1, notch receptor 2 and hairy/enhancer-of-split related with YRPW motif 1, which previously showed a negative regulation in myoblast differentiation. Conclusion: These findings provide a useful cell model for the future research on goose muscle development and suggest that CTRP3 may play an essential role in skeletal muscle growth of goose.

Effects of coated cysteamine hydrochloride on muscle fiber characteristics and amino acid composition of finishing pigs

  • Bai, Miaomiao;Liu, Hongnan;Xu, Kang;Yu, Rong;Oso, Abimbola Oladele;Deng, Jinping;Yin, Yulong
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.9
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    • pp.1430-1438
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    • 2019
  • Objective: This experiment was designed to determine the effects of coated cysteamine hydrochloride (CC) on muscle fiber characteristics, amino acid composition and transporters gene expression in the longissimus dorsi muscle (LDM) of finishing pigs. Methods: Two hundred and sixteen Duroc/Landrace/Yorkshire cross-bred male finishing pigs were fed with a corn-soybean basal diet supplemented with 0, 70, and 140 mg/kg cysteamine. Each group contained eight replicates of nine pigs per replicate. After 29 days, one pig was randomly selected from each replicate and slaughtered. Blood and LDM samples were collected and analyzed. Results: The results showed that supplemental dietary CC increased (p<0.05) the muscle fiber density. And CC supplementation also up-regulated (p<0.05) the expression of myosin heavy chain 1 (MyHC1) and MyHC2x mRNA levels, and down-regulated (p<0.05) MyHC2b expression in the LDM. Additionally, supplemental dietary CC reduced (p<0.05) the concentration of total cholesterol in the plasma and enhanced (p<0.05) the concentrations of essential amino acid and total amino acid in the LDM. The relative expression levels of chloramphenicol acetyltransferase 2, $b^{0,+}$ amino acid transporter, and $y^+$-L-type amino acid transporter 1 were upregulated (p<0.05) in the LDM when pigs were fed with the dietary CC of 70 mg/kg. Conclusion: Cysteamine supplementation could increase fiber density and distribution of fiber types. It also improved the deposition of protein in the LDM by up-regulated the expression of amino acid transporters.

Effects of Corn Silage on Growth Performance, Carcass Characteristics, and Gene Expression of Longissimus dorsi Muscle Biopsy of Hanwoo Steers (옥수수 사일리지가 거세한우의 성장, 도체특성 및 등심 유전자 발현에 미치는 영향)

  • Kim, U.H.;Kang, D.H.;Park, B.H.;Jang, S.S.;Chung, K.Y.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.24 no.1
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    • pp.41-48
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    • 2022
  • This study was conducted to investigate effects of corn silage on final fattening performance, carcass characteristics, and gene expression of Hanwoo Longissimus dorsi muscle biopsy. Twenty one steers with initial body weight of control 291.5±6.5kg, corn silage 291.7.0±17.3kg were used for 18 months of fattening period. Average daily gain of corn silage tended to increase compared to control in early fattening period(p=0.092). Feed conversion ratio of corn silage was higher than control in early fattening(p=0.005). The animals in corn silage increased A grade 23% in meat quantity than the control. Myogenic gene expression on the Longissimus dorsi biopsy were compared between corn silage and control. The level of myosin heavy chain(MHC) I, IIX mRNA were greater than the control in the whole period(p<0.05). The level of Peroxisome proliferator-activated receptor γ (PPAR γ) mRNA was greater than the corn silage(p<0.05). Inconclusion, corn silage will be possible to use as an alternative concentrate feeding system.

Anti-diabetic effects of the extract from Atractylodes lancea, Anemarrhena asphodeloides and Cinnamomum Cassia mixture in high fat diet-induced diabetic mice and regulation of the function in C2C12 mouse skeletal muscle cells (창출·지모·육계 복합추출물의 고지방식이 유도 당뇨병 마우스에서의 항당뇨 효능 및 C2C12 골격근세포에서의 조절기전 연구)

  • Park, Ki Ho;Kang, Seok Yong;Kang, Anna;Jung, Hyo Won;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.34 no.6
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    • pp.79-89
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    • 2019
  • Objective : This study investigated the anti-diabetic effects of DM1, a herbal mixture with Atractylodis Rhizoma, Anemarrhenae Rhizoma, and Cinnamomi Cortex in high fat diet (HFD)-induced diabetic mice and the mechanism in C2C12 mouse skeletal muscle cells. Methods : The C57B/6 mice were fed high fat for 12 weeks, and then administrated DM1 extract (500 mg/kg, p.o.) for 4 weeks. The changes of body weight, calorie and water intakes, fasting blood glucose levels and the serum levels of glucose, insulin, triglyceride, HDL-cholesterol, AST and ALT were measured in mice. The histological changes of liver and pancreas tissues were also observed by H&E stain. C2C12 myoblasts were differentiated into myotubes and then treated with DM1 extract (0.5, 1, and 2 mg/㎖) for 24 hr. The expression of myosin heavy chain (MHC), PGC1α, Sirt1 and NRF1, and the AMPK phosphorylation were determined in the myotubes by western blot, respectively. Results : The DM1 extract administration significantly decreased the calorie and water intakes, glucose, triglyceride, AST and ALT levels and increased insulin and HDL-cholesterol in HFD-induced diabetic mice. DM1 extract inhibited lipid accumulation in liver tissue and improved glucose tolerance. In C2C12 myotubes, DM1 treatment increased the expression of MHC, PGC1α, Sirt-1, NRF-1 and the AMPK phosphorylation. Conclusion : In our results indicate that DM1 can improve diabetic symptoms by decreasing the obesity, glucose tolerance and fatty liver in HFD-induced diabetic mice, and responsible mechanism is might be related with energy enhancement.

Effects of feeding high-energy diet on growth performance, blood parameters, and carcass traits in Hanwoo steers

  • Kang, Dong Hun;Chung, Ki Yong;Park, Bo Hye;Kim, Ui Hyung;Jang, Sun Sik;Smith, Zachary K.;Kim, Jongkyoo
    • Animal Bioscience
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    • v.35 no.10
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    • pp.1545-1555
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    • 2022
  • Objective: Our study aimed to investigate the effects of a 2% increase in dietary total digestible nutrients (TDN) value during the growing (7 to 12 mo of age) and fattening (13 to 30 mo of age) period of Hanwoo steers. Methods: Two hundred and twenty Hanwoo steers were assigned to one of two treatments: i) a control group (basal TDN, BTDN, n = 111 steers, growing = 70.5%, early fattening = 71.0%, late fattening = 74.0%) or high TDN (HTDN, n = 109 steers, growing = 72.6%, early = 73.1%, late = 76.2%). Growth performance, carcass traits, blood parameters, and gene expression of longissimus dorsi (LD) (7, 18, and 30 mo) were quantified. Results: Steers on the BTDN diets had increased (p≤0.02) DMI throughout the feeding trial compared to HTDN, but gain did not differ appreciably. A greater proportion of cattle in HTDN received Korean quality grade 1 (82%) or greater compared to BTDN (77%), while HTDN had a greater yield grade (29%) than BTDN (20%). Redness (a*) of LD muscle was improved (p = 0.021) in steers fed HTDN. Feeding the HTDN diet did not alter blood parameters. Steers fed HTDN diet increased (p = 0.015) the proportion of stearic acid and tended to alter linoleic acid. Overall, saturated, unsaturated, monounsaturated, and polyunsaturated fatty acids of LD muscle were not impacted by the HTDN treatment. A treatment by age interaction was noted for mRNA expression of myosin heavy chain (MHC) IIA, IIX, and stearoyl CoA desaturase (SCD) (p≤0.026). No treatment effect was detected on gene expression from LD muscle biopsies at 7, 18, and 30 mo of age; however, an age effect was detected for all variables measured (p≤0.001). Conclusion: Our results indicated that feeding HTDN diet could improve overall quality grade while minimum effects were noted in gene expression, blood parameters, and growing performance. Cattle performance prediction in the feedlot is a critical decision-making tool for optimal planning of cattle fattening and these data provide both benchmark physiological parameters and growth performance measures for Hanwoo cattle feeding enterprises.