• 대한한방내과학회지
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• 제29권1호
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• pp.243-257
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• 2008

Myostatin, a negative regulator of myogenesis, is shown to function by controlling the proliferation of myoblasts. In this study we show that myostatin is an inhibitor of myoblast differentiation and that this inhibition is mediated through Smad 3. To determine MyoD expression by Dodamtang treatment, we compared the expression pattern of $C_{2}C_{12}$ mouse myoblasts that constitutively express myostatin with control cells. In vitro, increasing concentrations of Dodamtang reversibly prevented the myogenic blockage of myoblasts by myostatin expression. ELISA assay, Western and confocal analysis indicated that treatment of Dodamtang to the low serum culture media increased the levels of MyoD leading to the inhibition of myogenic differentiation by myostatin. The stable transfection of $C_{2}C_{12}$ myoblasts with myostatin expressing constructs did rescue MyoD-induced myogenic differentiation. Consistent with this, the treatment of Dodamtang rescued the expression of a MyoD in $C_{2}C_{12}$ myoblasts treated with myostatin. Taken together, these results suggest that induction of MyoD by Dodamtang inhibits myostatin activity and expression via SMAD3 resulting in the rescue of the myoblasts to differentiate into myotubes. Thus we propose that myostatin action by Dodamtang plays a critical role in myogenic differentiation and that the muscular hyperplasia and hypertrophy seen in animals that blockage of functional myostatin is because of deregulated proliferation and differentiation of myoblasts.

• 대한본초학회지
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• 제23권2호
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• pp.159-168
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• 2008

Objectives : The present study has been undertaken to investigate the effects of Dipsaci Radix on Muscle Fiber Atrophy and MyoD Expression in Gastrocnemius of MCAO Rats Methods : In order to investigate effects of Dipsaci radix on the skeletal muscle atrophy following stroke, cerebral infarct was induced by the middle cerebral artery occlusion (MCAO) in the rats. Water extract of Dipsaci radix (184.4 mg/100 g) was treated for 4 weeks, once a day orally, after the MCAO. Effects were evaluated with muscle fiber type composition and cross-sectioned area of muscle fibers in gastrocnemius of the unaffected & affected hind limbs. And MyoD protein expression in gastrocnemius was demonstrated with immunohistochemistry and western blotting. Results : Obtained results were as follows; 1. Infarct volume was not attenuated by Dipsaci radix treatment in the MCAO rats. 2. At the affected-side hind limb of the MCAO rats, the increase of type-I fibers and the decrease of type-II fibers were induced by Dipsaci radix treatment. 3. At the affected-side hind limb of the MCAO rats, decreases of cross-sectioned areas of type-I and type-II fibers were attenuated by Dipsaci radix treatment. 4. At the affected-side hind limb of the MCAO rats, MyoD positive cells were increased by Dipsaci radix treatment. 5. At the affected-side hind limb of the MCAO rats, MyoD expressions were increased by Dipsaci radix treatment. Conclusions : These results suggest that Dipsaci radix has a protective effect against muscle atrophy, through the inhibition of the muscle cell apoptosis, following the central nervous system demage.

• 한국미생물·생명공학회지
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• 제46권2호
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• pp.102-110
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• 2018

myo-Inositol (MI)을 대사하여 다른 물질로 전환하는 미생물을 과수원 토양으로부터 분리하였다. 분리균 YB-46은 유일한 탄소원으로 MI이 첨가된 배지에서 성장하였고 16S rDNA 염기서열에 따라 Enterobacter 속의 균주로 추정되었다. Fosmid pCC1FOS 벡터를 사용하여 제조된 거대 유전체 은행으로부터 MI을 미지의 대사 물질로 전환하는 Escherichia coli 형질전환주를 선발하였다. 이로부터 플라스미드를 분리하고 삽입된 유전자의 일부 염기서열을 결정한 결과 336 아미노 잔기로 구성된 myo-inositol dehytrogenase (IolG)를 암호화하는 iolG 유전자가 발견되었다. 분리균 YB-46의 IolG는 E. aerogenes와 Bacillus subtilis의 IolG와 약 50% 수준의 상동성을 보였다. 카르복실 말단에 hexahistidine이 연결되도록 제조한 His-tagged IoG (HtIolG)의 유전자를 재조합 대장균에서 발현하여 균체 파쇄액으로부터 HtIolG를 정제하였다. 정제된 HtIolG는 $45^{\circ}C$와 pH 10.5에서 최대 활성을 보였고 MI과 D-glucose에 대한 활성이 가장 높았으며 D-chiro-inositol, D-mannitol 및 D-xylose에도 90% 이상의 활성을 보였다. 최적 반응조건에서 MI을 기질로 하여 반응 동력학적 계수를 측정한 결과 $K_m$과 $V_{max}$가 1.83 mM과 $0.724{\mu}mol/min/mg$로 확인되었다. HtIolG의 활성은 $Zn^{2+}$에 의해 1.7배 증가하였으며, $Co^{2+}$와 SDS에 의해서는 크게 감소하였다.

• Journal of Microbiology and Biotechnology
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• 제13권6호
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• pp.906-911
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• 2003

The function of genes related to spectinomycin biosynthesis (spcD, speA, speB, spcS2) from Streptomyces spectabilis ATCC 27741, a spectinomycin producer, was analyzed. Each gene was subcloned from a spectinomycin biosynthetic gene cluster and overexpressed in E. coli BL21 (DE3) using pET vector. After incubating each purified protein with its possible substrates, the final products were analyzed using high-performance liquid chromatography (HPLC). From these results, spcD, speA, and speB have been identified to be dTDP-glucose synthase, myo-inositol monophosphatase, and myo-inositol dehydrogenase, respectively. In addition, the results suggest that the spcS2 gene product functions downstream of the speB gene product in the biosynthetic pathway of spectinomycin. Taken together, the present study elucidates the early steps of the biosynthetic pathway for 6-deoxyhexose (6-DOH) part (actinospectose) and aminocyclitol part (actinamine) of spectinomycin.

• 한국미생물·생명공학회지
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• 제24권4호
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• pp.485-492
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• 1996

myo-Inositol, a growth factor for Saccharomyces cerevisiae (S. cerevisiae), has been known to be incorporated into phosphatidylinositol (PI), which is a kind of phospholipid in the cell membrane, by a membrane-associated PI-synthesizing enzyme. The deficiency of myo-inositol in S. cerevisiae adversely affected the membrane structure and function. On the basis of biochemical functions of myo-inositol, the effect of deficiency of myo-inositol on the aerobic glucose metabolism was investigated by measuring specific oxygen uptake rate (Q$_{O2}$) used as an indicator representing the respiratory capacity of S. cerevisiae in batch and continuous cultures. The respiratory capacity of aerobic glucose metabolism in S. cerevisiae was also monitored after glucose pulse-addition in a continuous culture (D=0.2, 1/hr), in which glucose was utilized through respiratory metabolism. The deficiency of myo-inositol was found to lead to both the decrease of the maximum specific oxygen uptake rate (Q$_{O2max}$) observed from the batch as well as in the continuous culture experiment and the decrease of the respiratory capacity of aerobic glucose metabolism of S. cerevisiae determined from the glucose pulse-addition experiment, in which the glucose flux into respiratory and fermen- tative metabolism was quantitatively analyzed.

• 한국축산식품학회지
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• 제41권4호
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• pp.623-635
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• 2021

The effect of deer antler extract on muscle differentiation and muscle atrophy were evaluated to minimize muscle loss following aging. Various deer antler extracts (HWE, hot water extract of deer antler; FE, HWE of fermented deer antler; ET, enzyme-assisted extract of deer antler; UE, extract prepared by ultrasonication of deer antler) were evaluated for their effect on muscle differentiation and inhibition of 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR)-induced muscle atrophy in C2C12 cells. Morphological changes according to the effect of antler extracts on muscle differentiation were confirmed by Jenner-Giemsa staining. In addition, the expression levels of genes related to muscle differentiation and atrophy were confirmed through qRT-PCR. In the presence of antler extracts, the length and thickness of myotubes and myogenin differentiation 1 (MyoD1) and myogenic factor 5 (Myf5) gene expression were increased compared to those in the control group (CON). Gene expression of AMP-activated protein kinase (AMPK), MyoD1, and myogenin, along with the muscle atrophy factors muscle RING finger-1 (MuRF-1) and forkhead box O3a (FoxO3a) upon addition of deer antler extracts to muscle-atrophied C2C12 cells was determined by qRT-PCR after treatment with AICAR. The expression of MuRF-1 and FoxO3a decreased in the groups treated with antler extracts compared to that in the group treated with AICAR alone. In addition, gene expression of MyoD1 and myogenin in the muscle atrophy cell model was significantly increased compared that into the CON. Therefore, our findings indicate that antler extract can increase the expression of MyoD1, Myf5 and myogenin, inhibit muscle atrophy, and promote muscle differentiation.

• Animal cells and systems
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• 제2권1호
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• pp.117-122
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• 1998

D-raf, a Drosophila homolog of the human c-raf-1, is known as a signal transducer in cell proliferation and differentiation. A previous study found that the D-raf gene expression is regulated by the DNA replication-related element (DRE)/DRE-binding factor (DREF) system. In this study, we found the sequences homologous to transcription factor C/EBP, MyoD, STAT and Myc recognition sites in the D-raf promoter. We have generated various base substitutional mutations in these recognition sites and subsequently examined their effects on D-raf promoter activity through transient CAT assays in Kc cells with reporter plasmids p5'-878DrafCAT carrying the mutations in these binding sites. Through gel mobility shift assay using nuclear extracts of Kc cells, we detected factors binding to these recognition sites. Our results show that transcription factor C/EBP, STAT and Myc binding sites in D-raf promoter region play a positive role in transcriptional regulation of the D-raf gene and the Myo D binding site plays a negative role.

• The Journal of Korean Physical Therapy
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• 제29권3호
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• pp.145-151
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• 2017

Purpose: The purpose of this study was to investigate the validity and reliability of the measurement of shoulder joint motions using an inertial measurement unit (IMU). Methods: For this study, 33 participants (32 females and 1 male) were recruited. The subjects were passively positioned with the shoulder placed at specific angles using a goniometer (shoulder flexion $0^{\circ}-170^{\circ}$, abduction $0^{\circ}-170^{\circ}$, external rotation $0^{\circ}-90^{\circ}$, and internal rotation $0^{\circ}-60^{\circ}$ angles). Kinematic data on the shoulder joints were simultaneously obtained using IMU three-dimensional (3D) angular measurement (MyoMotion) and photographic measurement. Test-retest reliability and concurrent validity were examined. Results: The MyoMotion system provided good to very good relative reliability with small standard error of measurement (SEM) and minimal detectable change (MDC) values from all three planes. It also presented acceptable validity, except for some of shoulder flexion, shoulder external rotation, and shoulder abduction. There was a trend for the shoulder joint measurements to be underestimated using the IMU 3D angular measurement system compared to the goniometer and photo methods in all planes. Conclusion: The IMU 3D angular measurement provided a reliable measurement and presented acceptable validity. However, it showed relatively low accuracy in some shoulder positions. Therefore, using the MyoMotion measurement system to assess shoulder joint angles would be recommended only with careful consideration and supervision in all situations.

• Asian-Australasian Journal of Animal Sciences
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• 제29권7호
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• pp.1037-1043
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• 2016

Epigenetic processes in the development of skeletal muscle have been appreciated for over a decade. DNA methylation is a major epigenetic modification important for regulating gene expression and suppressing spurious transcription. Up to now, the importance of epigenetic marks in the regulation of Pax7 and myogenic regulatory factors (MRFs) expression is far less explored. In the present study, semi-quantitative the real-time polymerase chain reaction (RT-PCR) analyses showed MyoD and Myf5 were expressed in activated and quiescent C2C12 cells. MyoG was expressed in a later stage of myogenesis. Pax7 was weakly expressed in differentiated C2C12 cells. To further understand the regulation of expression of these genes, the DNA methylation status of Pax7, MyoD, and Myf5 was determined by bisulfite sequencing PCR. During the C2C12 myoblasts fusion process, the changes of promoter and exon 1 methylation of Pax7, MyoD, and Myf5 genes were observed. In addition, an inverse relationship of low methylation and high expression was found. These results suggest that DNA methylation may be an important mechanism regulating Pax7 and MRFs transcription in cell myogenic differentiation.

• 한방재활의학과학회지
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• 제18권1호
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• pp.47-63
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• 2008

Objectives : Eucommiae cortex is reported that it helps bone and skeletal muscle stronger. In case of bone, many report is presented, but reports related to skeletal muscle are rarely existed. So in order to investigate effects of Eucommiae cortex on the skeletal muscle atrophy following stroke, cerebral infarct was induced by the middle cerebral artery occlusion (MCAO) in the rats. Methods : In order to induce MCAO rats, nylon suture was advanced and then blocked middle cerebral artery(MCA). Water extract of Eucommiae cortex was treated for 15 days, once a day orally, after the MCAO. Effects were evaluated with muscle weights, muscle fiber type composition, cross-sectioned area of muscle fibers in soleus and gastrocnemius of the unaffected and affected hind limbs. And MyoD protein expression in gastrocnemius was demonstrated with immunohistochemistry and western blotting. Results : In the affected hind limb of the MCAO rats, muscle weight loss of gastrocnemius and tibialis anterior muscles were attenuated by Eucommiae cortex treatment. In soleus muscle of the affected hind limb of the MCAO rats, increase of type-I fibers and decrease of type-II fibers were induced by Eucommiae cortex treatment. In soleus muscle of the affected hind limb of the MCAO rats, decrease of cross-sectioned areas of type-I fibers was attenuated by Eucommiae cortex treatment. In gastrocnemius muscle of the affected hind limb of the MCAO rats, increase of type-I fibers and decrease of type-II fibers were induced by Eucommiae cortex treatment. In gastrocnemius muscle of the affected hind limb of the MCAO rats, decreases of cross-sectioned areas of type-I and type-II fibers were attenuated by Eucommiae cortex treatment. In gastrocnemius muscle of the affected and unaffected hind limb of the MCAO rats, MyoD expressions were increased by Eucommiae cortex treatment. Conclusions : These results suggest that Eucommiae cortex has a protective effect against muscle atrophy, through the inhibition of the muscle cell apoptosis, following the central nervous system demage.