• Title/Summary/Keyword: cell-free extract

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Protective Effects of Membrane-Free Stem Cell Extract from H2O2-Induced Inflammation Responses in Human Periodontal Ligament Fibroblasts (무막줄기세포추출물의 H2O2에 의해 유도된 치주 세포의 염증 반응 보호 효과)

  • He, Mei Tong;Kim, Ji Hyun;Kim, Young Sil;Park, Hye Sook;Cho, Eun Ju
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.6
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    • pp.95-103
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    • 2019
  • Periodontal inflammation, a major kind of periodontal diseases, is characterized to bleed, pain, and teeth loss, and it is resulted from oxidative stress. Membrane-free stem cell extract could avoid the immunogencity rejection by removal of cell membrane. In the present study, we investigated the protective effect of membrane-free stem cell extract from oxidative stress-induced periodontal inflammation in human periodontal ligament fibroblasts (HPLF). In the cell viability measurement, membrane-free stem cell extract showed significant increase of cell viability, compared with the $H_2O_2$-treated control group. To further investigation of molecular mechanisms, we measured inflammation and apoptosis related protein expressions. Membrane-free stem cell extract attenuated inflammation-related protein expressions such as nuclear factor kappa light chain enhancer of activated B cells, inducible nitric oxide synthase, and interleukin-6. In addition, the treatment of membrane-free stem cell extract decreased apoptotic protein expressions such as cleaved caspase-9, -3, poly (ADP-ribose) polymerase, and B-cell lymphoma 2 (Bcl-2)-associated X protein/Bcl-2 ratio in the $H_2O_2$-treated HPLF cells. In conclusion, membrane-free stem cell extract exhibited anti-oxidative stress effects by regulation of inflammation and apoptosis in HPLF, suggesting that it could be used as the treatment agents for periodontal inflammatory disease.

Effect of Lactobacilli on Reactive Oxygen Scavenging and Immune Stimulation (유산균의 활성산소 소거 및 면역증강효과)

  • lee, Ho;Yang, Seung-Gak;Park, Soo-Nam;Jeon, Do-Yong
    • KSBB Journal
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    • v.7 no.4
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    • pp.290-295
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    • 1992
  • Reactive oxygen scavenging activity and immune stimulatory activity of lactobacilli were investigated by different free radical scavenging assays and Ig G assay. Lactobacilli culture (S/N) and its complex with $Mn^{2+}$ have significant effects in XOD assay and response to paraquat. Cell free extract significantly prevented the photohemolysis. Thus, it seems that each sample from lactobacilli has a different free radical scavening mechanism. Furthermore, it is assumed that cell free extract from lactobacilli activates antibody stimulation of B cell through a stimulation of T cell.

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The Effect of Alisma orientale Extract on Free Fatty Acid-induced Lipoapoptosis in HepG2 Cells (택사(澤瀉)가 유리지방산으로 유발된 HepG2 cell의 lipoapoptosis에 미치는 영향)

  • Kim, Eun-Young;Lee, Jang-Hoon
    • The Journal of Internal Korean Medicine
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    • v.35 no.2
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    • pp.184-194
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    • 2014
  • Objectives : This study was designed to investigate the effect on lipoapoptosis of Alisma orientale extract against free fatty acid-induced cellular injury. Methods : HepG2 cells were used in an vitro model. HepG2 cells were treated with free fatty acids to generate a cellular model of nonalcoholic fatty liver disease (NAFLD). Using this cellular model, the anti-apoptotic effect and reducing steatosis of Alisma orientale extract against free fatty acid-induced cellular injury was evaluated by measuring steatosis and apoptosis. Results : Alisma orientale extract significantly attenuated free fatty acid-induced intracellular steatosis. Alisma orientale extract inhibited free fatty acid-mediated activation of pJNK, PUMA, BAX, caspase-3, and -9, and apoptotic kinases that are correlated with NAFLD. Alisma orientale extract also promoted Bcl-2, a anti-apoptotic protein. Conclusions : From the above, the Alisma orientale extract decreased the hepatocyte steatosis and showed the hepatocelluar protective effect by the regulation of apoptosis-related protein. It proposes the possibility of Alisma orientale extract to the treatment of nonalcoholic fatty liver disease in clinics.

The Effects of BunSimGiEumGami-Bang(Fenxinqiyinjiameifang) on Serotonin of P815 cell (분심기음가미방(分心氣飮加味方)의 항산화능과 serotonin 대사에 미치는 영향)

  • Lim, Jae-Won;Jung, In-Chul;Lee, Sang-Ryong
    • Journal of Oriental Neuropsychiatry
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    • v.22 no.2
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    • pp.147-162
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    • 2011
  • Objectives : This experiment was designed to investigate the effect of the BSGE hot water extract on serotonin biosynthesis of depression model. Methods : The cytotoxicity of the BSGE hot water extract was analyzed by MTT assay on P815 cell. The antioxidant activity was measured by DPPH free radical-scavenging activity and SOD activity on P815 cell. The quantity of 5-HT and expression of TPH-1, AAADC and MAOa mRNA was measured by of HPLC profle Analysis on P815 cell. Results : 1. The BSGE hot water extract increased DPPH free radical-scavenging activity and SOD activity on P815 cell. 2. The BSGE hot water extract increased significantly the quantity of 5-HT. 3. The BSGE hot water extract increased the expression of TPH-1 mRNA. Conclusions : This experiment shows that the BSGE hot water extract might be effective for the prevention and treatment of depression. Investigation into the clinical use of the BSGE hot water extract for depression is suggested for future research.

Protective Effect of Acanthopanax senticosus Extract on Alloxan-induced β-cell Damage

  • Rho, Hye-Won;Lee, Ji-Hyun;Kim, Jong-Suk;Kim, Hyung-Rho;Park, Byung-Hyun;Park, Jin-Woo
    • Preventive Nutrition and Food Science
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    • v.10 no.1
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    • pp.46-51
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    • 2005
  • The protective effect of Acanthopanax senticosus (AS) extract on alloxan-induced pancreatic β-cell damage was investigated in HIT T-15 cells, a Syrian hamster pancreatic β-cell line. Alloxan caused the pancreatic β-cell damage through the generation of reactive oxygen free radicals, increased DNA fragmentation, and decreased cellular NAD/sup +/ levels. The β-cell damage was significantly prevented by the pretreatment with water soluble extract of AS roots. These results suggest that the protective effect of AS extract, on alloxan-induced β-cell damage, is primarily due to the inhibition of the generation of reactive oxygen free radical species (ROS) by alloxan.

Biosynthesis of $\beta$-Lactam Antibiotics by Cell-free Extract from Lysobacter lactamgenus

  • Roh, Ju-Won;Nam, Doo-Hyun
    • Archives of Pharmacal Research
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    • v.15 no.3
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    • pp.234-238
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    • 1992
  • Using cell-free extract of Lysobacter lactamgenus, enzymatic conversion of $\delta$-L-($\alpha$-aminoadiphyl)-L-cysteinyl-D-valine (ACV) the first substrate of $\beta$-lactam biosynthesis, into antibiotic compounds was attempted. In high performance liquid chromatographic (HPLC) analysis, the biosynthetic intermediates for cephalosporin antibiotics including isopenicillin N, deacetoxycephalosporin C, deacetylcephalosporin C and unknown cephem compound were detected in reaction mixtures. It implies that cephabacin compounds from L lactamgenus could be produced by biosynthetic routes through penicillin ring formation and its expansion to cephalosporin ring, likely as cephalosporin C from Cephalosporium or cephamycin C from Streptomyces. Among biosynthetic enzyme in cell-free extract, the ring formation activity (isopenicillin N synthetase activity) was separated in 50-60% of ammonium sulfate fraction, and ring expansion activity (deacetoxycephalosporin C synthetase activity) was found to be in 40-50% fraction. The partially purified isopenicillin N synthetase could convert as much as 90% ACV to isopenicillin N during 6-hour reaction.

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The Conversion of Lithocholic Acid into 5$\beta$-Androstan-3, 17-dione in the Cell-free System of Mycobacterium sp. NRRL B-3805

  • Lee, Kang-Man;Park, Hye-Kyung
    • Archives of Pharmacal Research
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    • v.14 no.3
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    • pp.261-265
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    • 1991
  • In a microbial cell-free extract system, side chain cleavage on various sterols and steroids was tested. The cell-free extracts of Mycobacterium sp. NRRL B-3805 showed the side chain cleavage activity on lithocholic acid to form 5$\beta$-androstan-3.17-dione. The properties of the activity were examined.

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Glucose Oxidation and It's Oxidative Enzyme Systems in Dunaliella tertiolecta. (II) Evidence for Glycolytic and Pentose Phosphate Pathways in Cell-free Extracts (Dunaliella tertiolecta의 포도당산화와 산화효소계 (II) Cell-free Extracts를 사용한 Glycolytic 및 Pentose Phosphate Pathway의 존재확인)

  • 권영명
    • Journal of Plant Biology
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    • v.12 no.2
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    • pp.15-22
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    • 1969
  • By spectrophotometric assay method, the following enzymes could be detected in Dunaliella tertiolecta and Chlorella pyrenoidosa cell-free extracts: Hexokinase; Glucose-6-phosphate, 6-Phosphogluconate and Triosephosphate dehydrogenase; Transketolase; Phosphogluco and Ribosephosphate isomerase; Phosphoglucomutase; Phosphofructokinase; Fructosediphosphate aldorase and Ribulosephosphate 3-epimerase. Such enzymes are in accordance with the proposed pathway of glucose catabolism by D. tertiolecta as well as C. pyrenoidosa. Also, it could be estimated, under the presence of NADP, that pentose phosphate pathway were more active than glycolytic pathway in D. tertiolecta cell-free systems.

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Enhancement of Glucose-Fueled Cell-Free Protein Synthesis by the Addition of Lipids (지질의 첨가를 통한 포도당 기반 무세포 단백질 합성 시스템의 단백질 발현 효율 향상)

  • Lee, So Jeong;Kim, Ho-Cheol;Kim, Dong-Myung
    • Korean Chemical Engineering Research
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    • v.57 no.1
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    • pp.85-89
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    • 2019
  • Cell-free protein synthesis utilizes the translational machinery in a cell extract. Unlike the conventional cell-based expression methods, not being affected by the conditions for cell growth, cell-free protein synthesis enables flexible manipulation of individual factors affecting the efficiency protein biosynthesis. However, the high cost and low stability of the energy sources to regenerate ATP have limited the use of cell-free synthesis for large-scale production of recombinant proteins. One of the approaches to address this problem is to use glucose as an alternative energy source to regenerate ATP through the glucose-metabolizing pathways in a cell extract. In this study, in an attempt to improve the efficiency of ATP regeneration by reinforcing oxidative phosphorylation process, we supplemented with cellular lipids to a glucose-fueled reaction mixture for cell-free protein synthesis. As a result of the lipid supplementation, the productivity of chloramphenicol acetyltransferase in a cell-free synthesis system using glucose increased more than 6 fold compared to when the lipid was not supplemented.

Producyion of Threonine Using Methanol Dehydrogenase and Serine Hydroxyltransferase in a New Methylotrophic Bacterium KJ29 (New Methylotrophic Bacterium KJ29의 Methanol Dehydrogenase와 Serine Hydroxymethyltransferase를 이용한 Threonine의 생산에 관한 연구)

  • 김경자
    • Microbiology and Biotechnology Letters
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    • v.21 no.6
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    • pp.577-581
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    • 1993
  • The amino acid threonine was produced from glycine and ethanol in a reaction mixture using cell free extract of the methylotrophic bacterium isolated from soil and identified as mellthylo-bacterium sp. KJ29. Although the isolate could grow on carbon source other than methanol, only the cell free extract from the cells grown on methanol produced threonine. Methanol dehydrogenase (MDH) activity was present only in the cells grown on methanol when compared to the cells grown on heterotrophic substrates.

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