• KSBB Journal
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• 제14권5호
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• pp.523-527
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• 1999

A naturally luminescent bacterium, Photobacterium phosphoreum was stored in 2.5% NaCl solution at 2$0^{\circ}C$, 4$^{\circ}C$, -2$0^{\circ}C$ and -7$0^{\circ}C$ for 30 days. In vivo luminescence and concentrations of total and culturable cells were determined by luminometer, spectrophotometer and dilution plate counting, respectively. When stored at 4$^{\circ}C$ and 2$0^{\circ}C$, concentrations of cells were rapidly decreased as a result of cell lysis, leading to adrop in turbidity and cultured counts. The bioluminescence of cells stored at 4$^{\circ}C$ was maintained until 12 days while those of cells starved at other temperatures decreased to background level within 3 days. Following incubation of stored cells in fresh liquid medium, activities of viable cells increased throughout storage period excepting cells stored at 2$0^{\circ}C$. Changes in bioluminescence intensity following addition of 2.5% NaCl solution markedly showed in cells stored at -2$0^{\circ}C$ and -7$0^{\circ}C$ and increased to maximum 8 fold.

• 한국식품저장유통학회지
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• 제31권2호
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• pp.298-306
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• 2024

본 연구에서는 식물 추출물 혼합 분말의 근육세포 분화 및 산화적 스트레스에 대한 세포 보호 효과를 확인하고자 하였다. 추출물을 최대 1,000 ㎍/mL 농도까지 세포에 처리한 결과, 세포 생존율이 감소하지 않음을 확인되었다. 식물 추출물 혼합분말이 근육세포 분화 인자에 미치는 영향을 확인하기 위해 myogenin과 MHC의 발현 여부를 확인한 결과, 무처리군에 비해 발현이 증가함을 확인하였다. H₂O₂에 의해 유도된 산화적 스트레스에 대한 세포 보호 효과를 확인한 결과, 식물 추출물 혼합 분말 처리에 의해 H₂O₂ 단독 처리군보다 세포 생존율이 증가하였으며, LDH와 creatine kinase의 활성이 감소하였다. 또한, Bax와 Bcl-2의 발현을 조절하여 caspase-9와 -3 활성화를 억제함을 확인하였다. 이를 통해 식물 추출물 혼합 분말의 근육세포 분화 효과 및 H₂O₂에 의해 유도된 산화적 스트레스에 대한 세포 보호 효과가 있음을 확인하였다. 따라서 식물 추출물 혼합 분말은 근감소증 개선을 위한 기능성 소재로써 활용이 가능할 것으로 판단되며, 향후 근감소증 개선을 위한 기능성 소재로서의 유효성 확보를 위해서 근감소증 세포 및 동물모델을 이용한 효능 및 기전 분석 연구가 더 필요할 것으로 사료된다.

• 생명과학회지
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• 제25권10호
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• pp.1098-1102
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• 2015

본 연구에서는 C2C12 근육 세포의 분화 과정에 있어 IGF-I이 지방산의 수송을 담당하는 FABPpm mRNA 및 단백질 발현에 어떠한 영향을 미치는지에 대해 알아보았다. 그 결과 근육세포의 분화에 있어 FABPpm의 단백질과 mRNA 발현이 IGF-I에 의해 유의하게 조절되었음을 알 수 있었다. 이는 기존의 여러 연구의 결과인 IGF-I이 골격근에서 근육 관련 유전자들의 발현을 조절하여 근부피 유지 및 증대에 중심적인 역할 것뿐만 아니라, 지방산의 수송을 담당하는 FABPpm의 발현에도 영향을 미친다는 사실을 밝혔다는 것에 의의가 있다고 생각된다. 향후 골격근에서 IGF-I에 의한 FABPpm 발현 조절에 있어, 세포 내부에서의 신호전달 경로 및 상호작용 인자들에 관한 연구를 통해 지방 대사를 조절하는 기전에 관한 연구가 필요할 것으로 사료된다.

• 생명과학회지
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• 제12권1호
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• pp.55-60
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• 2002

골격근 세포의 분화는 근육특이 유전자들의 전사적 활성과 근육아세포에서 근육소관으로의 형태적 분화로 특징지어진다. 본 연구에서는 TSA가 근육형성의 일련의 과정에서 NF-kB DNA 결합 활성과 융합에 미치는 영향을 조사하였다. 대조군과 비교해서 TSA가 처리된 C2C12 myoblast는 융합하여 근육소관을 형성할 수 없었으며 NF-kB DNA 결합 활성은 억제되었다. 이런 현상들이 TSA에 의한 직접적인 것인지 알아보기 위해서 TSA가 처리되지 않고 분화를 유도하기 위해서 사용된 배지를 농축하여 C2C12 myoblast에 TSA와 함께 동시에 처리하였다. 그 결과 세포는 융합하여 근육소관을 형성하였으며 NF-kB DNA 결합 활성이 회복되었다. 이러한 결과는 TSA가 아마도 여러 관련 인자들을 통해 myoblast의 융합과 NF-kB DNA 결합 활성을 억제함으로 근육형성과정에 영향을 미침을 시사한다.

• Journal of Oral Medicine and Pain
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• 제26권1호
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• pp.39-50
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• 2001

In order to investigate the effect of Transforming growth factor ${\beta}1$(below TGF-${\beta}1$) and osteogenic protein-1(below Op-1) onto the myogenic differentiation, C2C12 satellite myoblastic cell line was cultured and treated with both growth factors. At first morphological changes with microscopical examination were examined, and isolated total RNA to analyse mRNA expression of bone marker proteins, muscle regulatory proteins, TGF-${\beta}$ receptor and their ligands by Northern blot analysis. And cellular proliferative inducibility of both growth factors was also tested to C2C12 cells. Incubating the cell with $5ng/m{\ell}$ of TGF-${\beta}1$ until 4 days almost inhibited multinucleated myotube formation expressing muscular regulatory proteins, and induced decreasing Id proteins. However, no osteoblastic phenotypes was induced by TGF-${\beta}1$ in C2C12 cells. The mRNA expression of TGF-${\beta}$ receptors with TGF-${\beta}1$ was conversed after 48 hours cultured. Type I TGF-${\beta}$ receptor was seemed to play a role in negative signalling for inhibition of myogenic differentiation. OP-1 dose dependently induced ALP activity, osteopontine production and bone sialoprotein production at concentrations above $100ng/m{\ell}$ and osteocalcin production at concentrations above $300ng/m{\ell}$. The concentration of OP-1 required to induce these osteoblastic phenotypes was the same as that required to almost completely inhibit myotube formation. Incubation with above $100ng/m{\ell}$ OP-1 suppressed the expression of mRNA for muscular egulatory proteins from 2 days after incubation. Expression of Id-1, 2, 3 mRNA were stimulated by OP-1 at concentration above $300ng/m{\ell}$. When C2C12 cells were treated with both growth factors, TGF-${\beta}1$ potentiated the inhibitory effect of OP-1 on myotube formation and expression of mRNA for myogenin at 12 days. And TGF-${\beta}1$ reduced osteocalcin and bone sialoprotein production induced by OP-1 at 12 days in C2C12 cells. Both growth factor had no mitogenic effect. These results indicate that OP-1 converts the differentiation pathway of C2C12 myoblasts into that of osteoblastic lineage cells and it's not heritable, but TGF-${\beta}1$ does not and has reversible inhibitory activity on the myogenic differentiation. TGF-${\beta}1$ and OP-1 play a role in myogenic differentiation via different mechanism between them.

• 대한의생명과학회지
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• 제12권3호
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• pp.131-137
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• 2006

RGS is a negative regulator of G-protein signaling and can be identified by the presence of a conserved $120{sim}125$ amino acid motif, which is referred to as the RGS box. A number of RGSs are induced in response to a wide variety of stimuli. Increased levels of RGSs lead to significant decreases in GPCR responsiveness. To obtain further evidence of a role of RGS proteins in rat C6 astrocytoma cells, we first determined the expression profile of RGS-specific mRNA in C6 cells using reverse transcription-polymerase chain reaction (RT-PCR) with a poly dT18 primer and transcript-specific primers. We found that RGS2, RGS3, RGS6, RGS9, RGS10, RGS12, and RGS16 were differentially expressed in C6 astrocytoma cells. The highest expression rate was found for RGS3, followed by RGS16, RGS10 and RGS9, whereas the expression level for RGS2 was barely detectable. We next assessed whether forskolin regulated the expression of RGSs expressed in C6 astrocytoma cells. The present study found that forskolin dose-dependently stimulated the expression of RGS2 transcripts. This up-regulation of RGS2 gene was abrogated by H-89, potent and broad-spectrum protein kinase A (PKA) inhibitors. Actinomycin D completely inhibited the up-regulation of RGS2 gene induced by forskolin $(10{\mu}M)$, indicating that the regulation of RGS2 gene is controlled at the transcriptional level. In addition, forskolin did significantly activate transcriptional cAMP response element (CRE) in either HEK 293 cells or C6 cells and did not modulate the $NF-{\kappa}B$ and AP-l activity as measured by luciferase reporter gene assay. Finally, forskolin induced the expression of RGS2 mRNA in C6 astrocytoma cells, which depend on the PKA pathway and CRE transcriptional pathways.

• Nutrition Research and Practice
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• 제16권2호
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• pp.147-160
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• 2022

BACKGROUND/OBJECTIVES: Patients with chronic kidney disease (CKD) have a high concentration of uremic toxins in their blood and often experience muscle atrophy. Indoxyl sulfate (IS) is a uremic toxin produced by tryptophan metabolism. Although an elevated IS level may induce muscle dysfunction, the effect of IS on physiological concentration has not been elucidated. Additionally, the effects of ursolic acid (UA) on muscle hypertrophy have been reported in healthy models; however, it is unclear whether UA ameliorates muscle dysfunction associated with chronic diseases, such as CKD. Thus, this study aimed to investigate whether UA can improve the IS-induced impairment of mitochondrial biogenesis. MATERIALS/METHODS: C2C12 cells were incubated with or without IS (0.1 mM) and UA (1 or 2 μM) to elucidate the physiological effect of UA on CKD-related mitochondrial dysfunction and its related mechanisms using real-time reverse transcription-polymerase chain reaction, western blotting and enzyme-linked immunosorbent assay. RESULTS: IS suppressed the expression of differentiation marker genes without decreasing cell viability. IS decreased the mitochondrial DNA copy number and ATP levels by downregulating the genes pertaining to mitochondrial biogenesis (Ppargc1a, Nrf1, Tfam, Sirt1, and Mef2c), fusion (Mfn1 and Mfn2), oxidative phosphorylation (Cycs and Atp5b), and fatty acid oxidation (Pdk4, Acadm, Cpt1b, and Cd36). Furthermore, IS increased the intracellular mRNA and secretory protein levels of interleukin (IL)-6. Finally, UA ameliorated the IS-induced impairment in C2C12 cells. CONCLUSIONS: Our results indicated that UA improves the IS-induced impairment of mitochondrial biogenesis by affecting differentiation, ATP levels, and IL-6 secretion in C2C12 cells. Therefore, UA could be a novel therapeutic agent for CKD-induced muscle dysfunction.

• 대한한의학회지
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• 제25권3호
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• pp.149-159
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• 2004

Backgrounds & Objectives : The water extract of Gyungokgo (GOG) has traditionally been used for treatment of general weakness and hemoptysis in oriental medicine. However, little is known about the mechanism by which the water extract of GOG rescues cells from these damages. This study was designed to investigate the protective mechanisms of GOG on H2O2­induced cell death in H9c2 cardiomyoblasts. Methods : In this study, we used H9c2 cells. Cells were treated with oxidative stress in the absence and presence of 1000㎍/ml GOG for 12hrs. Cells were treated with various concentrations of GOG for 12hrs. Cell viability was measured by MTT assay. Oxidative stress, which markedly decreased the viability of H9c2 cells, was characterized by apparent apoptotic features such as chromatin condensation as well as fragmentation of genomic DNA and nuclei. Results : GOG significantly reduced H₂O₂-induced cell death and apoptotic characteristics. The cotreatment of GOG and H₂O₂ in H9c2 cells also induced the phosphorylation of ERKs in a time-dependent manner. Moreover, PD098059, a MEK1 (upstream activator of ERK) inhibitor attenuated the protective effect of GOG on H₂O₂-induced cytotoxicity in H9c2 cardiomyoblast cells. Conclusions : These results suggest that MEK/ERK pathways play important roles in the protective effects of GOG in H9c2 cells. Taken together, they suggest that the protective effects of the water extracts of GOG against oxidative damages may be mediated by the regulation of HO-1, Fas/FasL and Bcl-XS proteins.

• 생명과학회지
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• 제22권12호
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• pp.1651-1657
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• 2012

본 연구에서는 C2C12 근육 세포에서 IGF-I이 세포골격 연결 단백질인 plectin과 MACF1 유전자 발현에 미치는 영향에 대해 알아보았다. 그 결과 IGF-I이 plectin 유전자의 단백질과 mRNA 발현을 증가시켰으며, MACF1 mRNA 발현을 증가시켰음을 알 수 있었다. 이는 운동에 의해 근육에서 분비가 증가하는 IGF-I이 근육 관련 유전자들의 발현을 조절하여 근부피 유지에 영향을 미친다는 기존의 연구 결과들에서 더 나아가 골격근 구조 안정화 및 근수축 기전에 기여하는 plectin과 MACF1 유전자 발현에도 영향을 미친다는 사실을 증명하였다는데 의의가 있다고 사료된다. 향후 근수축 기전에 있어, 운동 형태, 근섬유의 종류에 따른 세포 골격 단백질의 역할 규명 및 조절자에 관한 연구가 더 수행된다면 운동이 골격근의 생리적 변화에 미치는 영향에 대한 추가적 정보를 제공할 수 있을 것이다.

• Journal of Microbiology
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• 제40권1호
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• pp.38-42
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• 2002

The strain KKI isolated from soil contaminated with polycyclic aromatic hydrocarbons was identified as Pseundomonas sp. based on analyses by MIDI and Biolog Identification System. Cellular and physiological responses of strain KKI to two-ring polycyclic aromatic hydrocarbon, naphthalene were evaluated using radiorespirometry, PLFAs and sequence analysis of Rieske-type iron sulfur center of dioxygenase. KKI was found to be able to rapidly mineralize naphthalene. Notably, KKI cells pregrown on phenanthrene were able to mineralize naphthalene much more rapidly than naphthalenepregrown cells. The total cellular fatty acids of KKI were comprised of eleven C-even and two C-odd fatty acids (fatty acids < 0.2% in abundance were not considered in this calculation). Lipids 12:0 2OH, 12:03 OH, 16:0, 18:1 6c, 18:0 increased for naphthalene-exposed cells, while lipids 18:1 7c1/15:0 ism 2OH, 17:0 cyclo, 18:1 7c, 19:0 cyclo decreased. Data from Northern hybridization using a naphthalene dixoygenase gene fragment cloned out from KKI as a probe provided the information that naphthalene dioxygenase gene was more highly expressed in cells grown on phenanthrene than naphthalene.