• 제목/요약/키워드: Conditioned medium

검색결과 173건 처리시간 0.022초

Methanol Extract of Cinnamomum cassia Represses Cellular Proliferation and Gli-mediated Transcription in PANC-1 Human Pancreatic Cancer Cells

  • Lee, Hwa Jin
    • Natural Product Sciences
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    • 제20권3호
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    • pp.170-175
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    • 2014
  • Twenty five methanolic plant extracts were investigated to determine the anticancer activity against sonic hedgehog (shh)/Gli signaling pathway dependent cancer, PANC-1 human pancreatic cancer cells, through three screening programs. All extracts were inspected their inhibitory properties on sonic hedgehog-conditioned medium (shh-CM) induced alkaline phosphatase (ALP) activity in C3H10T1/2 mouse mesenchymal stem cells to examine whether the plant extracts affect the shh/Gli signaling pathway. Next, plant extracts were screened the ability to suppress the cell proliferation of PANC-1 human pancreatic cancer cells. Finally, active plant extracts from the two screening systems were evaluated for the suppressive effect on Gli-mediated transcriptional activity in PANC-1 cells. Among active plants, Cinnamomum cassia suppressed Gli-mediated transcriptional activity leading to the down-regulated expression of Gli-target genes such as Gli-1 and Patched-1 (Ptch-1). This study provides the consideration for the important role of natural products in drug discovery process as well as the basis for the further analysis of active plant and potential identification of novel bioactive compounds as inhibitors of Gli and therapeutic candidates against shh/Gli signaling pathway dependent cancers.

Hematopoietic effect of deer antler extract fermented by Bacillus subtilis on murine marrow cells

  • Park, Yooheon;Choi, Hyeon-Son;Lee, Hyun-Sun;Suh, Hyung Joo
    • Nutrition Research and Practice
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    • 제9권5호
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    • pp.451-458
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    • 2015
  • BACKGROUND/OBJECTIVES: We examined the chemical composition and the effect of fermented deer antler on hematopoietic factors in bone marrow cells. MATERIALS/METHODS: For the preparation of fermented deer antler extract (FAB), fermentation was carried out using Bacillus subtilis at $30^{\circ}C$ for 7 days. The hematopoietic effect of FAB was investigated hematopoietic factors in marrow cells. RESULTS: The contents of total sugar, sulfated glycosaminoglycans, and uronic acid and the dry weight gradually increased with fermentation time. The sialic acid content (from 0.14 mg/mL to 0.54 mg/mL) was the highest on the 4th day of fermentation after which it decreased. The proliferating activity of bone marrow cells increased with fermentation times. The levels of various hematopoietic growth factors were determined to verify the beneficial effect of deer antler extract fermented by B. subtilis on hematopoiesis. FAB increased the number of stem cell factors and granulocyte colony-stimulating factor in bone marrow cells. In addition, FAB augmented the burst-forming unit erythroid and total colonies in splenocyte-conditioned medium compared with non-fermented antler extract (NFA). However, FAB did not affect the mRNA levels of erythropoietin, an important factor for erythropoiesis. CONCLUSIONS: FAB, like NFA, did not directly affect hematopoiesis, but contributed to hematopoiesis by stimulating the production of hematopoietic factors.

GDNF secreted by pre-osteoclasts induces migration of bone marrow mesenchymal stem cells and stimulates osteogenesis

  • Yi, Sol;Kim, Jihee;Lee, Soo Young
    • BMB Reports
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    • 제53권12호
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    • pp.646-651
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    • 2020
  • Bone resorption is linked to bone formation via temporal and spatial coupling within the remodeling cycle. Several lines of evidence point to the critical role of coupling factors derived from pre-osteoclasts (POCs) during the regulation of bone marrow-derived mesenchymal stem cells (BMMSCs). However, the role of glial cell-derived neurotrophic factor (GDNF) in BMMSCs is not completely understood. Herein, we demonstrate the role of POC-derived GDNF in regulating the migration and osteogenic differentiation of BMMSCs. RNA sequencing revealed GDNF upregulation in POCs compared with monocytes/macrophages. Specifically, BMMSC migration was inhibited by a neutralizing antibody against GDNF in pre-osteoclast-conditioned medium (POC-CM), whereas treatment with a recombinant GDNF enhanced migration and osteogenic differentiation. In addition, POC-CM derived from GDNF knock-downed bone marrow macrophages suppressed BMMSC migration and osteogenic differentiation. SPP86, a small molecule inhibitor, inhibits BMMSC migration and osteogenic differentiation by targeting the receptor tyrosine kinase RET, which is recruited by GDNF into the GFRα1 complex. Overall, this study highlights the role of POC-derived GDNF in BMMSC migration and osteogenic differentiation, suggesting that GDNF regulates bone metabolism.

Physicochemical Analysis of Yogurt Produced by Leuconostoc mesenteroides H40 and Its Effects on Oxidative Stress in Neuronal Cells

  • Lee, Na-Kyoung;Lim, Sung-Min;Cheon, Min-Jeong;Paik, Hyun-Dong
    • 한국축산식품학회지
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    • 제41권2호
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    • pp.261-273
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    • 2021
  • Leuconostoc mesenteroides H40 (H40) was isolated from kimchi, and its probiotic properties and neuroprotective effect was evaluated in oxidatively stressed SH-SY5Y cells. H40 was stable in artificial gastric conditions and can be attached in HT-29 cells. In addition, H40 did not produce β-glucuronidase and showed resistant to several antibiotics. The conditioned medium (CM) was made using HT-29 cells refined with heat-killed probiotics (Probiotics-CM) and heated yogurts (Y-CM) to investigate the neuroprotective effect. Treatment with H40-CM not only increased cell viability but also significantly improved brain derived neurotropic factor (BDNF) expression and reduced the Bax/Bcl-2 ratio in oxidatively stress-induced SH-SY5Y cells. Besides, probiotic Y-CM significantly increased BDNF mRNA expression and decreased Bax/Bcl-2 ratio. The physicochemical properties of probiotic yogurt with H40 was not significantly different from the control yogurt. The viable cell counts of lactic acid bacteria in control and probiotic yogurt with H40 was 8.66 Log CFU/mL and 8.96 Log CFU/mL, respectively. Therefore, these results indicate that H40 can be used as prophylactic functional dairy food having neuroprotective effects.

Neuroprotective Effects of Heat-Killed Levilactobacillus brevis KU15152 on H2O2-Induced Oxidative Stress

  • Hyun-Ji Bock;Na-Kyoung Lee;Hyun-Dong Paik
    • Journal of Microbiology and Biotechnology
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    • 제33권9호
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    • pp.1189-1196
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    • 2023
  • This study proposed to demonstrate the neuroprotective effects of heat-killed Levilactobacillus brevis KU15152. Heat-killed L. brevis KU15152 showed antioxidant activity similar to that of Lacticaseibacillus rhamnosus GG, in terms of radical scavenging activity. To evaluate the neuroprotective effects, conditioned medium (CM) obtained by incubating heat-killed bacteria in intestinal cells (HT-29) was used through gut-brain axis. CM from L. brevis KU15152 protected neuroblastoma cells (SH-SY5Y) against H2O2-induced oxidative stress. Pretreatment with CM significantly alleviated the morphological changes induced by H2O2. Heat-killed L. brevis KU15152 showed an increased brain-derived neurotrophic factor (BDNF) expression in HT-29 cells. L. brevis KU15152-CM remarkably downregulated the Bax/Bcl-2 ratio, while upregulating the expression of BDNF and tyrosine hydroxylase (TH) in SH-SY5Y cells. Furthermore, L. brevis KU15152-CM reduced caspase-3 activity following H2O2 treatment. In conclusion, L. brevis KU15152 can be potentially used as food materials to avoid neurodegenerative diseases.

Oral squamous carcinoma cells stimulated by Porphyromonas gingivalis-derived lipopolysaccharide induce osteoclastogenesis through a paracrine mechanism

  • Bo Ram Keum;Soon Chul Heo;Hyung Joon Kim
    • International Journal of Oral Biology
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    • 제49권3호
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    • pp.79-86
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    • 2024
  • Periodontal disease (PD) is strongly linked to increased risk of oral squamous cell carcinoma (OSCC); however, the specific mechanism through which the development of PD and OSCC is simultaneously promoted remains unclear. This study explored the impact of periodontal pathogens on OSCC progression and the contribution of periodontal pathogen-stimulated OSCC to PD development. The expression of osteoclastogenesis-inducing factors was assessed using quantitative reverse transcription polymerase chain reaction analysis following stimulation of OSCC with lipopolysaccharide (LPS) derived from the periodontal pathogen Porphyromonas gingivalis (Pg), a pathogen commonly responsible for PD. The cell counting kit-8 assay was used to determine the effects of Pg-LPS on OSCC proliferation and drug resistance to cisplatin and 5-fluorouracil. The effects of conditioned medium (CM) derived from Pg-LPS-stimulated OSCC on osteoclastogenesis was evaluated using tartrate-resistant acid phosphatase (TRAP) staining on bone marrow-derived macrophages (BMMs). Pg-LPS administration in SCC-25 and YD-8 OSCC cell lines induced a significant increase in receptor activator of nuclear factor kappa-B ligand mRNA expression; however, it did not affect cell proliferation. Treatment with CM derived from Pg-LPS-stimulated SCC-25 or YD-8 cells markedly enhanced the formation of TRAP-positive multinucleated cells during osteoclast differentiation of BMMs. Altogether, these findings demonstrate that Pg-LPS-stimulated OSCC promoted osteoclastogenesis through a paracrine mechanism.

Sucrose, 칼슘, 붕소 첨가가 복숭아 화분 발아에 미치는 영향 (Effect of Sucrose, Calcium and Boron Added in the Medium on Pollen Germination of Peach(Prunus persica SIEB))

  • 천병덕;최인수;강점순
    • 생명과학회지
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    • 제16권4호
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    • pp.561-565
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    • 2006
  • 우리나라 과수 중 개화기가 빠른 복숭아는 저온에 의하여 화분발아 장해로 결실율이 저하되고 있다. 본 연구는 저온에서 화분 발아율을 증진시킬 수 있는 방법을 강구하기 위한 기초연구로서 화분 발아조건과 배지내에 sucrose, $CaCl_2$$H_3BO_3$ 첨가농도를 구명하고자 일련의 실험을 실시하였다. 복숭아 화분은 배지에 치상한 후 30분부터 발아가 시작되어 3시간 30분까지 지속되었으나, 그 이후는 더 이상 발아되지 않았다. 따라서 복숭아의 화분발아 판정을 위한 적정 시간은 치상 후 4시간이 좋았다. 복숭아 화분의 적정발아 온도는 $20^{\circ}C$였고, 이보다 저온이거나 고온에서는 화분발아율이 감소하였다. Sucrose는 화분발아를 촉진하였고 최적농도는 10%였다. 무기염류 중 $H_3BO_3$ 첨가는 '장택백봉'과 '수봉' 품종에서 화분 발아율을 무처리에 비해 $20{\sim}30%$ 증진시켰으며, 그 효과는 저온인 $10^{\circ}C$에서 현저하였다. 최적처리 농도는 $100\;mg\;{\cdot}\;L^{-1}$ 처리였다. 반면 $CaCl_2$ 첨가하면 화분발아율이 감소하였다. $10^{\circ}C$의 저온에서 화분발아율은 품종간에 뚜렷한 차이가 있었으며, '장택백봉', '백봉', '월미', '백도', '대구보', '유명', '수봉', '백미조생'의 순으로 화분발아율이 낮았다.

Conjugated Linoleic Acid (CLA)와 그 이성체가 전립선 암세포의 증식에 미치는 영향 (The Effect of Conjugated Linoleic Acid and Its Isomers on the Proliferation of Prostate TSU-Prl Cancer Cells)

  • 오윤신;김은지;김종우;김우경;이현숙;윤정한
    • Journal of Nutrition and Health
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    • 제35권2호
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    • pp.192-200
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    • 2002
  • Conjugated linoleic acid (CLA) is a collective term for positional and geometric isomers of octadecadienoic acid in which the double bonds are conjugated. CLA has anticancer activity in a variety of animal cancer models, and cis-9, trans-11 (c9t11) and trans-10, cis-12(t10c12) CLA are the most predominant isomers present in the synthetic preparations utilized in these animal studies. To compare the ability of c9t11, t10c12 and an isomeric mixture of CLA to inhibit TSU-Prl cell growth, cells were incubated in a serum-free medium with various concentrations of these fatty acids. The isomeric mixture inhibited cell growth in a dose-dependent manner (1-3 $\mu$M) with a 41 $\pm$ 1% inhibition observed at 3 $\mu$M concentration after 48 hours. T10c12 also inhibited cell proliferation in a dote-dependent manner, However, the efficacy and potency of this isomer was much greater than that of the isomeric mixture with a 49 $\pm$ 2% inhibition observed at 0.3 $\mu$M concentration after 48 hours. By contrast, c9t11 slightly increased cell proliferation. To determine whether the growth-inhibiting effect of CLA is related to the changes in production of insulin-like growth factors (IGF) and IGF-binding proteins (IGFBP) by these cells, serum-free conditioned media were collected. Immunoblot analysis of conditioned media using a monoclonal anti-IGF-II antibody showed that both the isomeric mixture and t10c12 inhibited secretion of both mature 7,500 Mr and higher Mr forms of pro IGF-II, whereas c9t11 had no effect. Ligand blot analysis with 125I-IGF-II revealed the presence of two types of IGFBPs : 24,000 Mr IGFBP-4 and 30,000 Mr IGFBP-6. The production of IGFBP-4 slightly decreased at the highest concentrations of the isomeric mixture and t10c12. These results indicate that CLA inhibits human prostate cancer cell growth, an effect largely due to the action of t10c12. The growth inhibition may result, at least in part, from decreased production of IGF-II and IGFBP-4 by these cells.

혈소판농축혈장 내의 성장요소가 조골세포주의 활성도에 미치는 영향 (THE EFFECT OF GROWTH FACTORS IN PLATELET-RICH PLASMA ON THE ACTIVITY OF OSTEOBLAST CELL LINE)

  • 정태욱;김영수;김창회;장경수;임영준
    • 대한치과보철학회지
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    • 제42권2호
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    • pp.175-191
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    • 2004
  • Statement of problem: Platelet-rich plasma(PRP) is well known to be very effective method to stimulate and accelerate the healing of bone and soft tissue. However, there are few reports which deal with the mechanisms of the PRP on the activation of the osteoblasts. Purpose: This study was aimed to investigate the effect of growth factors in PRP on the activity of osteoblasts. Material and method: To evaluate the effect on human, human osteoblast cell line was cultured. PRP was extracted from the blood of a healthy volunteer. Using the recombinant growth factors of PDGF, $TGFT-\beta$, IGF-1, bFGF which are mainly found at bone matrix and their neutralizing antibody, the effect of PRP on the attachment and proliferation of osteoblasts was evaluated. To evaluate the autocrine and paracrine effects, conditioned media(CM) of PRP was made and compared with PRP. By the western blot analysis, the expression of growth factors in PRP, CM was examined. Cell morphology was compared by the light microscope. Results : 1) The effects of CM on osteoblast were similar to the effects of PRP. 2) PRP, CM, recombinant $TGF-\beta$, bFGF, IGF-1 showed significantly higher cellular attachment than control(p<0.05) in the cell attachment assay. In the cell proliferation assay, PRP, CM, recombinant $TGF-\beta$, IGF-1, bFGF, PDGF increased significantly cell proliferation(p<0.01). Among the recombinant growth factors, IGF-1 showed the highest cellular attachment and proliferation. 3) In the western blot assay, bFGF, IGF-1, PDGF weve equally expressed in PRP and CM. 4) The attachment of osteoblast cell decreased significantly after the addition of neutralizing antibody against $TGF-\beta$, IGF-1(p<0.05). In the cell proliferation assay, the addition of neutralizing antibody against $TGF-\beta$, bFGF, PDGF, IGF-1 decreased significantly the cellular proliferation(p<0.05). The amount of decreasing in the cell attachment and proliferation is the highest in at-lGF-1. 5) The cells in control group were flattened and elongated with a few cellular processes in the a light microscope. But, the cells appeared as spherical, plump cells with well developed cellular processes in experimental groups. The cells in PRP and CM had more prominent developed features than recombinant growth factor groups. Conclusions : These findings imply that PRP maximize the cellular activity in early healing period using the synergistic effect, autocrine, paracrine effects of growth factors and increase the rate and degree of bone formation.

APPLICATION OF HF COASTAL OCEAN RADAR TO TSUNAMI OBSERVATIONS

  • Heron, Mal;Prytz, Arnstein;Heron, Scott;Helzel, Thomas;Schlick, Thomas;Greenslade, Diana;Schulz, Eric
    • 대한원격탐사학회:학술대회논문집
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    • 대한원격탐사학회 2006년도 Proceedings of ISRS 2006 PORSEC Volume I
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    • pp.34-37
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    • 2006
  • When tsunami waves propagate across open ocean they are steered by Coriolis force and refraction due to gentle gradients in the bathymetry on scales longer than the wavelength. When the wave encounters steep gradients at the edges of continental shelves and at the coast, the wave becomes non-linear and conservation of momentum produces squirts of surface current at the head of submerged canyons and in coastal bays. HF coastal ocean radar is well-conditioned to observe the current bursts at the edge of the continental shelf and give a warning of 40 minutes to 2 hours when the shelf is 50-200km wide. The period of tsunami waves is invariant over changes in bathymetry and is in the range 2-30 minutes. Wavelengths for tsunamis (in 500-3000 m depth) are in the range 8.5 to over 200 km and on a shelf where the depth is about 50 m (as in the Great Barrier Reef) the wavelengths are in the range 2.5 - 30 km. It is shown that the phased array HF ocean surface radar being deployed in the Great Barrier Reef (GBR) and operating in a routine way for mapping surface currents, can resolve surface current squirts from tsunamis in the wave period range 20-30 minutes and in the wavelength range greater than about 6 km. There is a trade-off between resolution of surface current speed and time resolution. If the radar is actively managed with automatic intervention during a tsunami alert period (triggered from the global seismic network) then it is estimated that the time resolution of the GBR radar may be reduced to about 2 minutes, which corresponds to a capability to detect tsunamis at the shelf edge in the period range 5-30 minutes. It is estimated that the lower limit of squirt velocity detection at the shelf edge would correspond to a tsunami with water elevation of less than 5 cm in the open ocean. This means that the GBR HF radar is well-conditioned for use as a monitor of small and medium scale tsunamis, and has the potential to contribute to the understanding of tsunami genesis research.

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