• Title/Summary/Keyword: C2C12 cell

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Structure of Cholesteryl Hemisuccinate (Cholesteryl Hemisuccinate의 구조)

  • Park, Young-Ja
    • Korean Journal of Crystallography
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    • v.15 no.1
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    • pp.29-34
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    • 2004
  • The crystal structure of cholesteryl hemisuccinate ($C_{31}H_{50}O_4$) was investigated by X-ray diffraction method. The cholesterol fragment of the title compound were in good agreement with those for related cholesterol derivatives. The unit cell contains four cholesteryl hemisuccinate molecules that are not related by crystal symmetry and have their tetracyclic system almost parellel to each other. There are two pairs of hydrogen-bonded dimer between A and D molecules and B and C molecules. These two hydrogen-bonded dimers are parallel to the c-axis, and are closely packed.

Immunoenhancing Effects of Conjugated Linoleic Acid on Chemotactic Activity of Porcine Peripheral Blood Polymorphonuclear Cells (돼지 말초혈액 다형핵 백혈구의 유주성에 있어서 conjugated linoleic acid의 면역증강효과)

  • Kim, Ju-hyang;Chung, Chung-soo;Lee, Chul-young;Yang, Mhan-pyo
    • Journal of Veterinary Clinics
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    • v.20 no.1
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    • pp.1-6
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    • 2003
  • Immunoenhancing effects of conjugated linoleic acid (CLA) isomers (l0t-l2c CLA, 9c-11t CLA, CLA mixture, 9c-11c CLA and 9t-11t CLA) on chemotactic activity of porcine peripheral blood polymorphonuclear cells (PMN) were examined. The chemotactic activity of PMN was evaluated by a modified Boyden chamber assay. CLA isomers at higher concentration of 50 to 200$\mu$M exhibited a low viability of cells by trypan blue exclusion. CLA isomers were used at concentration of 20uM showing no cytotoxic effect and high cell viability. CLA isomers themselves were not active or slight chemotactic for PMN. But culture supernatant from mononuclear cells (MNC) treated with 10t-12c CLA, 9c-11t CLA and CLA mixture except for 9c-11c. CLA and 9t-11t CLA enhanced remarkably chemotactic activity or porcine PMN PMN migration by culture supernatant from MNC treated with CLA mixture was found to be true chemotaxis by checkboard assay. This migration was also induced by porcine recombinant interleukin (rIL)-8. PMN chemotaxis caused both culture supernatant from MNC treated with CLA mixture and porcine rIL-8 was inhibited in a dose-dependent manner by addition of anti-porcine IL-8 polyclonal antibody. Therefore, these results strongly suggested that CLA (10t-12c CLA, 9c-11t CLA and CLA mixture) could stimulate porcine MNC to release and IL-8 like chemotactic activity.

Effects of polysaccharide (polycan) derived from black yeast in dexamethasone-induced muscle atrophy cell model (Dexamethasone으로 유도한 근위축 세포모델에서 흑효모 배양물 유래 polycan의 근위축 개선에 대한 효과)

  • Hwang, Su-Jin;Lim, Jong-Min;Ku, Bon-Hwa;Cheon, Da-Mi;Jung, Yu Jin;Kim, Young-Suk;Oh, Tae Woo
    • Herbal Formula Science
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    • v.29 no.1
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    • pp.45-55
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    • 2021
  • Objectives : This study was conducted to evaluate the anti-atrophic effect of polycan in dexamethasone-induced skeletal muscle atrophy in vitro model. Methods : C2C12 myoblast were differentiated into myotube by 2% horese serum medium for 6 days, and then treated polycan extract at different concentrations for 24h. The effect of dexamethasone on the induction of muscle atrophy and expression of atrophy-related genes in differentiated C2C12 myotubes using a GSH, ROS, real-time PCR, western blots analysis. Results : The results showed that Treatment with polycan (100 and 200 ㎍/㎖) noncytotoxic levels on both myoblast and myotube. Polycan decreased the ROS level overproduced with dexamethasone and improved the depletion of GSH level. Dexamethasone showed a decrease in myotube diameter, which was associated with up-regulation muscle-specific ubiquitin ligases markers, such as atrogin-1, FoxO3, myostatin and muscle RING finger-1 (MuRF1), and down-regulation of myogenin, MEF2, Myogenic regulatory factor 5, 6 and MyoD. The results showed that polycan treatment significantly dose-dependently inhibited it. Furthermore, decreased expressions of PI3K/Akt signal pathway by dexamethasone were reversed by treatment with polycan. Conclusions : Thus, polycan suppresses dexamethasone induced muscle atrophy in C2C12 myotube in vitro model through activation of PI3K/Akt pathway and protective effect of improve skeletal muscle function.

Analysis of Genes Regulated by HSP90 Inhibitor Geldanamycin in Neurons

  • Yang, Young-Mo;Kim, Seung-Whan;Kwon, O-Yu
    • Biomedical Science Letters
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    • v.15 no.1
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    • pp.97-99
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    • 2009
  • Geldanamycin is a benzoquinone ansamycin antibiotic that binds to cytosol HSP90 (Heat Shock Protein 90) and changes its biological function. HSP90 is involved in the intracellular important roles for the regulation of the cell cycle, cell growth, cell survival, apoptosis, angiogenesis and oncogenesis. To identify genes expressed during geldanamycin treatment against neurons of rats (PC12 cells), DNA microarray method was used. We have isolated 2 gene groups (up-or down-regulated genes) which are geldanamycin differentially expressed in neurons. Granzyme B is the gene most significantly increased among 204 up-regulated genes (more than 2 fold over-expression) and Chemokine (C-C motif) ligand 20 is the gene most dramatically decreased among 491 down-regulated genes (more than 2 fold down-expression). The gene increased expression of Cxc110, Cyp11a1, Gadd45a, Gja1, Gpx2, Ifua4, Inpp5e, Sox4, and Stip1 are involved stress-response gene, and Cryab, Dnaja1, Hspa1a, Hspa8, Hspca, Hspcb, Hspd1, Hspd1, and Hsph1 are strongly associated with protein folding. Cell cycle associated genes (Bc13, Brca2, Ccnf, Cdk2, Ddit3, Dusp6, E2f1, Illa, and Junb) and inflammatory response associated genes (Cc12, Cc120, Cxc12, Il23a, Nos2, Nppb, Tgfb1, Tlr2, and Tnt) are down-regulated more than 2 times by geldanamycin treatment. We found that geldanamycin is related to expression of many genes associated with stress response, protein folding, cell cycle, and inflammation by DNA microarray analysis. Further experimental molecular studies will be needed to figure out the exact biological function of various genes described above and the physiological change of neuronal cells by geldanamycin. The resulting data will give the one of the good clues for understanding of geldanamycin under molecular level in the neurons.

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Anti-diabetic mechanism of melania snail (Semisulcospira libertina) protamex hydrolysates (다슬기 protamex 가수분해물(MPH)의 항당뇨 기작 연구)

  • Pyo, Sang-Eun;Choi, Jae-Suk;Kim, Mi-Ryung
    • Food Science and Preservation
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    • v.24 no.7
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    • pp.1007-1016
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    • 2017
  • Melania snail (Semisulcospira libertina) was traditionally used as the healthy food in Korea. It was generally known to improve liver function and heal a diabetes. The aim of this study was to elucidate the anti-diabetic mechanism of melanian snail hydrolysates treated with protamex (MPH) by investigating the inhibitory action on protein tyrosine phosphatase 1B (PTP1B), the improving effect on the insulin resistance in C2C12 myoblast and the protective effect for pancreatic beta-cell (INS-1) under the glucose toxicity. The melania snail hydrolysates treated with protamex (MPH), which showed the highest degree of hydrolysis (43%), and inhibited effectively PTP1B activity ($IC_{50}=15.42{\pm}1.1{\mu}g/mL$), of which inhibitory effect was higher than usolic acid, positive control ($IC_{50}=16.65{\mu}g/mL$). MPH increased the glucose uptake in C2C12 myoblast treated with palmitic acid. In addition, MPH increased insulin mRNA expression level by over 160% with enhanced cell viability in INS-1 cell under the high glucose concentration (30 mM). These results suggest that MHP may improve the diabetic symptom by the inhibiting the PTP1B activity, increasing the glucose uptake in muscle cell and protecting the pancreatic beta-cell from glucose toxicity.

The Effect of Cultured Perichondrial Cell Sheet Covered Highly Active Engineered Cartilage: in vivo Comparative Assessment (배양연골막이 피복된 고효능 인공연골의 생체내 효과)

  • Park, Se-Il;Moon, Young-Mi;Jeong, Jae-Ho;Jang, Kwang-Ho;Ahn, Myun-Hwan
    • Journal of Veterinary Clinics
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    • v.28 no.5
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    • pp.486-496
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    • 2011
  • A special mesenchymal tissue layer called perichondrium has a chondrogenic capacity and is a candidate tissue for engineering of cartilage. To overcome limited potential for chondrocyte proliferation and re-absorption, we studied a method of cartilage tissue engineering comprising chondrocyte-hydrogel pluronic complex (CPC) and cultured perichondrial cell sheet (cPCs) which entirely cover CPC. For effective cartilage regeneration, cell-sheet engineering technique of high-density culture was used for fabrication of cPCs. Hydrogel pluronic as a biomimetic cell carrier used for stable and maintains the chondrocytes. The human cPCs was cultured as a single layer and entirely covered CPC. The tissue engineered constructs were implanted into the dorsal subcutaneous tissue pocket on nude mice (n = 6). CPC without cPCs were used as a controls (N = 6). Engineered cartilage specimens were harvested at 12 weeks after implantation and evaluated with gross morphology and histological examination. Biological analysis was also performed for glycosaminoglycan (GAG) and type II collagen. Indeed, we performed additional in vivo studies of cartilage regeneration using canine large fullthickness chondrial defect model. The dogs were allocated to the experimental groups as treated chondrocyte sheets with perichondrial cell sheet group (n = 4), and chondrocyte sheets only group (n = 4). The histological and biochemical studies performed 12 weeks later as same manners as nude mouse but additional immunofluorescence study. Grossly, the size of cartilage specimen of cPCs covered group was larger than that of the control. On histological examination, the specimen of cPCs covered group showed typical characteristics of cartilage tissue. The contents of GAG and type II collagen were higher in cPCs covered group than that of the control. These studies demonstrated the potential of such CPC/cPCs constructs to support chondrogenesis in vivo. In conclusion, the method of cartilage tissue engineering using cPCs supposed to be an effective method with higher cartilage tissue gain. We suggest a new method of cartilage tissue engineering using cultured perichondrial cell sheet as a promising strategy for cartilage tissue reconstruction.

The Crystal and Molecular Structure of Salicylaldehyde-4-piperidinothiosemicarbazone (Salicylaldehyde-4-piperidinothiosemicarbazone의 결정 및 분자구조)

  • Young-Ja Lee
    • Journal of the Korean Chemical Society
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    • v.20 no.1
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    • pp.3-14
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    • 1976
  • The crystal structure of alicylaldehyde-4-piperidinothiosemicarbazone, $C_{13}H_{l7}N_3OS$, has been determined by single crystal X-ray analysis. The crystals are orthorhombic, space group $P2_12_12_1$, with unit cell dimensions a = 6.52(2), b = 13.42(4), c = 14.92(4)${\AA}$. There are four formular units in a unit cell. The structure was solved by the heavy atom method and refined by isotropic block diagonal least-squares methods to a final R value of 0.10 for 1019 observed reflections. The oxygen atom of the hydroxyl group is involved in two hydrogen bonds, one as donor in the intramolecular O-H${\cdots}$N hydrogen bond and the other as acceptor in the intermolecular N-H${\cdots}$O hydrogen bond, the distances of the hydrogen bonds 2.56 and 3.00${\AA}$ respectively.The molecules are joined into infinite columns by the N-H${\cdots}o$O hydrogen bonds which form spirals along the two fold screw axis parallel to the a axis. The molecular columns are held together by van der Waals forces.

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The Crystal Structure of Benzidine Dihydrochloride (Benzidine 鹽酸鹽의 結晶構造)

  • Koo, Chung-Hoe;Kim, Hoon-Sup;Shin, Hyun-So
    • Journal of the Korean Chemical Society
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    • v.16 no.1
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    • pp.18-24
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    • 1972
  • Benzidine dihydrochloride crystallizes in the triclinic system. The space group is $P_1$. The unit cell dimensions are; a = 4.38${\pm}$0.01, b = 5.76${\pm}$0.01, c = 12.82${\pm}$0.02${\AA}$, $\alpha$ = 101.5${\pm}$0.2, $\beta$ = 99.5${\pm}$0.2, $\gamma$ = 99.5${\pm}$0.2$^{\circ}$; with one molecule per unit cell. The crystal structure has been solved by two dimensional Patterson and by trial and error methods, and refined by means of two dimensional differential synthesis. The bond distances are C-C(*) = 1.40${\pm}$0.02, C-C = 1.52${\pm}$0.02, C-N = 1.51${\pm}$0.03 and N-H${\cdot}{\cdot}{\cdot}$Cl = 3.21${\pm}$0.03${\AA}$. The structure consists of hydrogen bonded molecular layers, extending to the (100) plane, and the hydrogen bonding scheme is similar to that of p-phenylenediamine dihydrochloride. The adhesion between hydrogen bonded molecular layers is due to van der Waals forces.

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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.

Methylation of the Mouse Dlx5 and Osx Gene Promoters Regulates Cell Type-specific Gene Expression

  • Lee, Ji Yun;Lee, Yu Mi;Kim, Mi Jin;Choi, Je Yong;Park, Eui Kyun;Kim, Shin Yoon;Lee, Sam Poong;Yang, Jae Sup;Kim, Dong Sun
    • Molecules and Cells
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    • v.22 no.2
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    • pp.182-188
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    • 2006
  • Dlx5 and Osx are master regulatory proteins essential for initiating the cascade leading to osteoblast differentiation in mammals, but the mechanism of osteoblast-specific expression is not fully understood. DNA methylation at CpG sequences is involved in tissue and cell type-specific gene expression. We investigated the methylation status of Dlx5 and Osx in osteogenic and nonosteogenic cell lines by methylationspecific PCR (MSP). The CpG dinucleotides of the Dlx5 and Osx promoter regions were unmethylated in osteogenic cell lines transcribing these genes but methylated in nonosteogenic cell lines. Treatment of C2C12 cells with 5-AzadC induced dose- and timedependent expression of Dlx5 and Osx mRNA by demethylating the corresponding promoters. Furthermore the mRNAs for the osteoblast markers ALP and OC, which were undetectable in untreated cells, gradually increased after 5-AzadC treatment. In addition, BMP-2 stimulation induced Dlx5 expression by hypomethylating its promoter. These findings suggest that DNA methylation plays an important role in cell type-specific expression of Dlx5 and Osx.