• 제목/요약/키워드: cell-free extract

검색결과 583건 처리시간 0.038초

Effects of Salviae Miltiorrhizae Radix Hot Aqueous Extract on Nitric Oxide and Prostaglandin E2 Production and on 1,1-diphenyl-2-picryl hydrazyl Radical Scavenging in Macrophages

  • Yeo, In Ho;Lee, Cham Kyul;Lee, Eun Yong
    • 대한약침학회지
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    • 제17권1호
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    • pp.7-12
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    • 2014
  • Objectives: The objective of this study is to investigate the effects of Salviae Miltiorrhizae Radix hot aqueous extract on nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) production and on 1,1-diphenyl-2-picryl hydrazyl (DPPH) free-radical scavenging in macrophages. Methods: Salviae Miltiorrhizae Radix (300 g) was heated at $100^{\circ}C$ with distilled water (2 L) for 4 hours. The extract was filtered and concentrated to 100 mL by using a rotary evaporator, was frozen at $-80^{\circ}C$, and was then freeze-dried by using a freezing-drying system. The RAW 264.7 macrophage was subcultured by using $10-{\mu}g/mL$ lipopolysaccharide (LPS). In order to evaluate cytotoxicity, we performed 3-(4,5-dimrthylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays and measured the cell viability. The NO production was measured by using Griess assays, and the $PGE_2$ production was measured by using enzyme immunoassays. The antioxidant activity, the 1,1-diphenyl-2-picryl hydrazyl (DPPH) free-radical scavenging capability, was measured by using the DPPH method. Results: Cell viability with the 1-, 5-, 25-, 125- and $625-{\mu}g/mL$ Salviae Miltiorrhizae Radix hot aqueous extract was not significantly decreased compared to the cell viability without the extract. When 125 and $625{\mu}g/mL$ of Salviae Miltiorrhizae Radix hot aqueous extract were used, nitric oxide (NO) production in LPS-stimulated RAW 264.7 macrophages was significantly inhibited compared to that in the control group. When 25, 125, and $625{\mu}g/mL$ of Salviae Miltiorrhizae Radix hot aqueous extract were used, $PGE_2$ production in LPS-stimulated RAW 264.7 macrophages was significantly inhibited compared to that in the control group. The 125- and $625-{\mu}g/mL$ Salviae Miltiorrhizae Radix hot aqueous extracts had high DPPH free-radical scavenging capabilities in RAW 264.7 macrophages. Conclusion: This study indicates that Salviae Miltiorrhizae Radix hot aqueous extract suppresses NO and $PGE_2$ production and improves DPPH free-radical scavenging capability. Thus, it seems that Salviae Miltiorrhizae Radix hot aqueous extract may have an anti-inflammation effect and antioxidant activity.

Multifuctional Activities of Cultured extracts from Lactobacillus plantarum Ml as cosmeceutical ingredients.

  • S. Y. Vi;Lee, J. I;E. J. Han;G. J. Jung
    • 대한화장품학회:학술대회논문집
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    • 대한화장품학회 2003년도 IFSCC Conference Proceeding Book II
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    • pp.243-244
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    • 2003
  • The effects of Lactic acid bacteria have been investigated on anti-tumor. cholesterol reduction in blood. promotion of immune and skin-beauty. We are focused on cosmeceutical activity of Lactic acid bacteria (LAB). Ml, which is found in Korean traditional food. Kimchi The LAB.Ml has been identified as Lactobacillus plantarum Ml and individually cultured with Soybean soup and Soybean-Curd whey, until the total acidity has been reached the highest. After then, cell-free extracts from Ml have been used for the following studies. We assessed the effect of Lactobacillus plantarum Ml on the depigmentation of B16FlO melanoma cell. The melanin content of cells was decreased with 1-3% of cultured extracts. The tyrosinase activity was reduced by cell-free extracts of Lactobacillus plantarum Ml. Anti-aging and anti-oxidative activity of Ml cultured extract was also studied in NIH-3T3 human fibroblast cells. It showed that induction of cell proliferation. collagen synthesis and free radical scavenging activity. Additional studies for anti-fungal and anti-acne activity were also detected on Staphylococcus aureus and Propionibacterium acnes, respectively. These results suggest that cultured extract of Lactobacillun plantarum Ml would be used for cosmeceutical ingredients through multifunctional reaction on skin such as whitening, anti-wrinkle. anti-oxidation and anti-acnes.

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Antiproliferative Effects of Free and Encapsulated Hypericum Perforatum L. Extract and Its Potential Interaction with Doxorubicin for Esophageal Squamous Cell Carcinoma

  • Amjadi, Issa;Mohajeri, Mohammad;Borisov, Andrei;Hosseini, Motahare-Sadat
    • 대한약침학회지
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    • 제22권2호
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    • pp.102-108
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    • 2019
  • Objectives: Esophageal squamous cell carcinoma (ESCC) is considered as a deadly medical condition that affects a growing number of people worldwide. Targeted therapy of ESCC has been suggested recently and required extensive research. With cyclin D1 as a therapeutic target, the present study aimed at evaluating the anticancer effects of doxorubicin (Dox) or Hypericum perforatum L. (HP) extract encapsulated in poly(lactic-co-glycolic acid) (PLGA) nanoparticles on the ESCC cell line KYSE30. Methods: Nanoparticles were prepared using double emulsion method. Cytotoxicity assay was carried out to measure the anti-proliferation activity of Dox-loaded (Dox NPs) and HP-loaded nanoparticles (HP NPs) against both cancer and normal cell lines. The mRNA gene expression of cyclin D1 was evaluated to validate the cytotoxicity studies at molecular level. Results: Free drugs and nanoparticles significantly inhibited KYSE30 cells by 55-73% and slightly affected normal cells up to 29%. The IC50 of Dox NPs and HP NPs was ~ 0.04-0.06 mg/mL and ~ 0.6-0.7 mg/mL, respectively. Significant decrease occurred in cyclin D1 expression by Dox NPs and HP NPs (P < 0.05). Exposure of KYSE-30 cells to combined treatments including both Dox and HP extract significantly increased the level of cyclin D1 expression as compared to those with individual treatments (P < 0.05). Conclusion: Dox NPs and HP NPs can successfully and specifically target ESCC cells through downregulation of cyclin D1. The simultaneous use of Dox and HP extract should be avoided for the treatment of ESCC.

소평탕(消平湯)이 RIN-m5F 세포에서 인슐린 분비 및 유전자 발현에 미치는 영향 (Effect of Sopyung-tang Extract on Insulin Secretion and Gene Expression in RIN-m5F Cells)

  • 윤성식;조충식
    • 대한한방내과학회지
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    • 제31권1호
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    • pp.25-39
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    • 2010
  • Background : At high glucose levels in $\beta$-cells, cell viability and insulin secretion are decreased by glucotoxicity. Sopyung-tang(SPT) had an effect on blood glucose level decrease and antioxidant enzyme activities in streptozotocin-induced diabetic rats. Objectives : This study performed a series of experiment to verify the effects of SPT extract on the cell viability, antioxidant enzyme activities, insulin secretion and insulin mRNA expression at hyperglycemic states of RIN-m5F. Methods : After treatment at various concentrations of SPT added to the RIN-m5F cells, cell viability by MTT assay, free radical-scavenging activity, SOD activity and insulin secretion were measured. Additionally, insulin-related gene expression was measured using real-time RT-PCR. Results : Compared to the control group, SPT extract showed considerable effects on RIN-m5F cell viability, DPPH radical-scavenging activity, superoxide dismutase (SOD) activity, insulin secretion and insulin-related gene expression. Conclusions : This study showed that SPT extract has an effect on $\beta$-cell cell viability, insulin secretion and insulin-related gene expression. Thus, SPT extract may be used for treatment of diabetes and its complications. Further mechanism studies of SPT seem to be necessary on the glucotoxicity and oxidative stress.

Degradation of Polyvinyl Alcohol by Brevibacillus laterosporus: metabolic Pathway of Polyvinyl Alcohol to Acetate

  • Lim, Joong-Gyu;Park, Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • 제11권6호
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    • pp.928-933
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    • 2001
  • Approximately 0.1 mg/ml of polyvinyl alcohol (PVA) was degraded by the growing cell, Brevibacillus laterospours, for 30 h, and 0.2 mg/ml of PVA was degraded by the cell-free extract that was isolated from Brevibacillus laterosporus. Approximately $0.29{\mu}g$/ml of acetic acid was produced from PVA by using the cell-free extract as a catalyst for 40 min. $V_{max}\;and\;K_m$ value of purified PAV-degradation enzyme was 3.75g/l and 2.75 g/l/min in reaction with EDTA and 3.99 g/l and 2.98 g/l/min in reaction without EDTA, respectively. Molecular weight of the purified enzyme determined by SDS-PAGE was 63,000 Da. Alcohol dehydrogenase and aldehyde dehydrogenase activities were qualitatively detected on a native acrylamide gel by an active staining method, indicating the existence of the metabolic pathway to use PVA as a substrate.

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분자 샤페론을 사용한 연속확산식 무세포단백질 발현 시스템에서의 재조합 Plasminogen Activator의 효율적 발현 (Enhanced Synthesis of Active rPA in the Continuous Exchange Cell-free Protein Synthesis [CECF] System utilizing Molecular Chaperones)

  • 박창길;김태완;최차용;김동명
    • KSBB Journal
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    • 제21권2호
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    • pp.118-122
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    • 2006
  • 본 연구에서는 연속확산식 무세포 단백질 발현 시스템을 이용한 이황화결합 함유 단백질의 생산 시, 발현된 단백질의 응집으로 인한 비활성화의 문제점이 분파샤페론을 통한 발현 단백질의 용해도 증가, 세포 파쇄액의 화학적 처리를 통한 환원활성 제거, 산화환원완충액을 이용한 적절한 산화환원전위의 제공 등을 통해 단백질의 폴딩에 유리한 반응조건을 구축함으로써 해결될 수 있음을 보였다. 이러한 연구 결과는 각종 게놈 시퀀싱 프로젝트의 빠른 진척에 따라 현재 요구되고 있는 고속, 고효율의 단백질 발현 수단으로써 무세포 단백질 발현 시스템이 적용될 수 있는 가능성을 높여 줄 수 있을 것으로 기대된다.

지방간 유도 세포모델에서 황금(黃芩), 목향(木香), 시호(柴胡) 복합 추출물의 이상지질혈증에 대한 효과 (Anti-hyperlipidemic Effects of Scutellariae Radix, Aucklandiae Radix and Bupleuri Radix (SAB) extract in FL83B cells)

  • 도현주;김경호;오태우
    • 대한본초학회지
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    • 제35권5호
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    • pp.23-31
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    • 2020
  • Objectives : This study was conducted to evaluate the anti-hyperlipidemia effect of Scutellariae Radix, Aucklandiae Radix and Bupleuri Radix(SAB). Methods : FL83B cells were mouse liver hepatocytes, and we used this cell line. FL83B cells were treated with 0.5 mM oleic acid(OA) for 24 h, SAB extract was treated. After OA treatment, intracellular triglyceride (TG) and free fatty acid contents were measured with AdiopoRed™ assay and Free Fatty Acid Quantitation assay kit, respectively. Further, we evaluated several lipogenesis and metabolic markers such as sterol regulatory element-binding transcription factor-1c (SREBP-1c), acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), 3-hydroxy3-methyl-glutaryl CoA reductase (HMGCR), hormone-sensitive lipase (HSL), carnitine palmitoyltransferase (CPT-1), peroxisome proliferator activated receptor alpha (PPARα), and cluster of differentiation (CD36) using RT-PCR and Western-blot analysis. Results : OA markedly increased intracellular TG and free fatty acid, which plays a key role in reducing hepatic lipid accumulation, in FL83B cells. These increases were alleviated by SAB extract. The mRNA and protein expression of Fatty acid(FA) oxidation factors (CPT-1, PPARα), lipolysis factor(HSL), FA transporter(CD36), cholesterol synthesis factors (HMGCoA) and Lipodenesis (SREBP-1c, FAS, and ACC-1) were significantly increased by treatment of SAB extract in the OA-induced fatty liver cell model. Conclusions : In summary, the treat of SAB extract showed a significant reduction of the influx of fatty acids into hepatocytes, promoted the oxidation of fatty acids, and regulated fat synthesis-related factors, thereby regulating the accumulation of TG and free fatty acids.

음양곽(淫羊藿)이 토끼의 수축혈관에 미치는 이완효과 (Relaxation Effect of Epimedium Koreanum Extract on Rabbit Carotid Artery)

  • 노병진;박선영
    • 동의생리병리학회지
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    • 제27권6호
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    • pp.730-737
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    • 2013
  • The purpose of this study is to investigate vasorelaxant effect of Epimedium koreanum(EK) extract on rabbit carotid artery. In this study, to determine vasorelaxant effect of EK extract on rabbit carotid artery, arterial rings with intact or damaged endothelium were used for experiment using organ bath, and were contracted by norepinephrine(NE). After being contracted, arterial rings were treated with EK extract in a dose-dependent manner To study its mechanism, the contracted arterial rings induced by NE were pretreated with indomethacin(IM), $N_{\omega}$-nitro-L-arginine(L-NNA), methylene blue(MB) or tetraethylammonium chloride(TEA) and 0.1 $mg/m{\ell}$ EK extract was added. To analyze the effect of the EK extract on influx of extracellular calcium chloride($Ca^{2+}$) in rabbit carotid artery, in $Ca^{2+}$-free krebs solution, krebs solution containing 1 mM $Ca^{2+}$ was infused into the contracted arterial ring by NE after pretreatment of EK extract. To measure the cytotoxicity of the EK extract, cell viability of human umbilical vein endothelial cell(HUVEC) was measured by MTT assay, and nitric oxide(NO) was measured by Griess reagent. The EK extract significantly was relaxed the arterial ring with intact endothelium contracted by NE, but the vasorelaxant effect of the EK extract was inhibited in the arterial rings with damaged endothelium. The vasorelaxant effect of the EK extract was not different between the IM-pretreatedand and non-treated arterial rings. The vasorelaxant effect of EK extract were significantly inhibited, when arterial rings were pretreated with L-NNA, TEA, MB. And in $Ca^{2+}$-free krebs solution, increasing of arterial contraction by $Ca^{2+}$ addition were also inhibited by the treatment of EK, but not significant. The treatment of EK extract was increased NO concentration in HUVEC. This study suggested that the vasorelaxant effect of EK extract would be related with EDHF and NO production and increasing of cyclic GMP.

Inhibitory Effects of Latilactobacillus curvatus BYB3 Cell-Free Extract on Human Melanoma B16F10 Cells and Tumorigenic Mice

  • Dingyun Li;Xing Wang;Dong-June Park;Dong Hun Lee;Sejong Oh
    • Journal of Microbiology and Biotechnology
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    • 제34권3호
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    • pp.589-595
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    • 2024
  • Latilactobacillus curvatus BYB3 (BYB3) is a species of lactic acid bacteria, formerly named Lactobacillus curvatus, which is isolated from kimchi. In this study, the effect of BYB3, Lactobacillus rhamnosus GG, and Lactobacillus acidophilus GP1B strain extracts at various concentrations was examined on B16F10, a mouse melanoma cell line. Cell viability was examined via MTT assay, and the results indicated that compared to the other two probiotics, BYB3 significantly decreased the total percentages of viable cells. The effects of BYB3 on cell migration and proliferation in B16F10 cells were evaluated using wound healing mobility and proliferation assays, respectively; the results indicated that BYB3 inhibits cell migration and proliferation in a concentration-dependent manner. Using human dermal fibroblast cells to investigate BYB3 extract in vivo had no effect on skin-related cells. Nonetheless, the BYB3 extract inhibited tumor growth in a mouse model, as demonstrated by liver slices. Therefore, this suggests that using BYB3 extract to inhibit melanoma may be a novel approach.

Antioxidative activity of peony root

  • Lee, Seung-Chul;Kwon, Yong-Soo;Kim, Hyun-Pyo;Heo, Moon-Young
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 2003년도 Annual Meeting of KSAP : International Symposium on Pharmaceutical and Biomedical Sciences on Obesity
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    • pp.61-61
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    • 2003
  • The ethanol extract of peony root (Paeonia Lactiflora Pall, Paeoniaceae) and its major active components including gallic acid and methyl gallate were evaluated for their protective effects against free radical generation and lipid peroxidation. And protective effects against hydrogen peroxide-induced oxidative DNA damage in a mammalian cell line were performed. The ethanol extract of peony root (PRE), gallic acid and methyl gallate were shown to possess the significant free radical scavenging effect against 1,1-diphenyl-2-picryl hydrazine (DPPH) radical generation and were revealed the inhibitory effect of lipid peroxidation as expressed by malondialdehyde (MDA) formation. They were also found to strongly inhibit hydrogen peroxide-induced DNA damage from NIH/3T3 fibroblasts, assessed by single cell gel electrophoresis. Furthermore, oral administration of 50% PRE (50% ethanol extract), gallic acid and methyl gallate potently inhibited micronucleated reticulocyte (MNRET) formation of mouse peripheral blood induced by KBrO3 treatment in vivo. Therefore, PRE containing gallic acid and methyl gallate may be a useful natural antioxidant by scavenging free radicals, inhibition of lipid peroxidation and protecting oxidative DNA damage.

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