• Title/Summary/Keyword: HSC-T6

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Glial Fibrillary Acidic Protein Splice Variants in Hepatic Stellate Cells - Expression and Regulation

  • Lim, Michelle Chin Chia;Maubach, Gunter;Zhuo, Lang
    • Molecules and Cells
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    • v.25 no.3
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    • pp.376-384
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    • 2008
  • The glial fibrillary acidic protein (GFAP) is traditionally used as a marker for astrocytes of the brain, and more recently for the hepatic stellate cells (HSCs) of the liver. Several GFAP splice variants have been previously reported in the astrocytes of the CNS and in the non-myelinating Schwann cells of the PNS. In this study, we investigate whether GFAP splice variants are present in the HSCs and their expression as a function of HSCs activation. Furthermore, the regulation of these transcripts upon treatment with interferon gamma ($IFN-{\gamma}$) will be explored. Using semi-quan-titative RT-PCR and real-time PCR, we examine the expression and regulation of GFAP splice variants in HSCs as well as their respective half-life. We discover that most of the GFAP splice variants ($GFAP{\alpha}$, ${\beta}$, ${\delta}$, ${\varepsilon}$ and $\kappa$) found in the neural system are also expressed in quiescent and culture-activated primary HSCs. Interestingly, $GFAP{\alpha}$ is the predominant form in quiescent and culture-activated primary HSCs, while $GFAP{\beta}$, predominates in the SV40-immortalized activated HSC-T6. $GFAP{\delta}$, ${\varepsilon}$ and ${\kappa}$ have similar half-lives of 10 hours, while $GFAP{\beta}$ has a half-life of 17 hours. Treatment of HSC-T6 with $IFN-{\gamma}$ results in a significant 1.29-fold up-regulation of $GFAP{\alpha}$ whereas the level of the other transcripts remains unchanged. In summary, $GFAP{\alpha}$, ${\beta}$, ${\delta}$, ${\varepsilon}$ and $\kappa$ are present in HSCs. They are differentially regulated on the transcription level, implying a role of the 5' and 3' untranslated regions.

Inhibitory Effect of Angelica Gigantis Radix on Fibrogenesis in Rat Hepatic Stellate Cells (당귀(當歸)가 간성상세포의 섬유화 기전 억제에 미치는 영향)

  • Son, Hyun-Soo;Kim, Young-Chul
    • The Journal of Internal Korean Medicine
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    • v.31 no.3
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    • pp.415-424
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    • 2010
  • Objectives : This study was performed to investigate the anti-fibrogenic effect of Angelica Gigantis Radix on cultured rat hepatic stellate cells. Materials and Methods : Hepatic stellate cells(HSC-T6) were treated with various concentrations of Angelica Gigantis Radix extract for both 24 and 48 hours. The extraction was done either with distilled water or 80% EtOH. After the treatment, cell viability, cell proliferation, procollagen production and the mRNA expression of the ASMA, TIMP1, TIMP2, procollagen Type 1a2, and Cytokine IL-6 production were measured by using MTT assay, BrdU assay, RT-PCR, procollagen Type I C-peptide EIA and IL-6 ELISA assay. Results : The cell viability treated with water extraction was significantly increased, but there were no significant changes treated with 80% EtOH extraction. The cell proliferation treated with water extraction decreased only in the 24 hours group, while there were significant decreases either in the 24 and 48 hours groups treated with 80% EtOH extraction. The mRNA expressions of the ASMA, TIMP2 and procollagen 1a2 decreased in a concentration-dependent manner in the 48 hours group. Procollagen production decreased in a concentration-dependent manner in both the 24 and 48 hours groups. Cytokine IL-6 production increased in a concentration-dependent manner in both the 24 and 48 hours groups. Conclusion : These results suggest that Angelica Gigantis Radix is beneficial in the treatment of cirrhotic patients as well as for patients with chronic hepatitis.

Inhibitory Effect of Artemisiae Capillaris Herba on Fibrogenesis in Rat Hepatic Stellate Cells (인진(茵蔯)이 간성상세포의 섬유화 억제에 미치는 영향)

  • Kim, Seong-A;Woo, Hong-Jung;Kim, Young-Chul;Lee, Jang-Hoon
    • The Journal of Internal Korean Medicine
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    • v.29 no.1
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    • pp.177-188
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    • 2008
  • Objectives : This study was performed to investigate the anti-fibrogenic effect of Artemisiae Capillaris Herba on cultured rat hepatic stellate cells. Materials and Methods : Hepatic stellate cells(HSC-T6) were treated with various concentrations of Artemisiae Capillaris Herba extract for 24 hours. The extraction was done either with distilled water or 50% EtOH. After the treatment, cell viability, proliferation, procollagen levels and the mRNA of the collagen type 1a2 and ASMA were measured by using MTT assay, BrdU assay, RT-PCR, and Procollagen Type I C-peptide EIA Kit. Results : The viability and proliferation of the hepatic stellate cells were decreased as the concentration increased. The mRNA expression decreased consistently with the volume of the secreted procollagen with the extraction made with distilled water, which indicates the herb has inhibitory effect on fibrogenesis of the liver by regulating one of the fibrosis associated genes in transcription. However, it increased in 50% EtOH extraction, which shows that a more stable reaction is expected of the extraction made with distilled water than the extraction made with 50% EtOH. The production of procollagen was decreased by a low-concentration treatment with Artemisiae Capillaris Herba, but increased by a high concentration. It seemed that the cells were responding to Artemisiae Capillaris Herba in low- concentrations, thus producing small amounts of collagen. When the drug was administered at high enough concentration to give direct toxicity to cells, the ability of cells to produce collagen was activated, and the overproduction of collagen was observed as an undesirable results. Conclusion : These results suggest that Artemisiae Capillaris Herba is beneficial in the treatment of cirrhotic patients as well as for the patients with chronic hepatitis when extracted with water in the proper concentrations.

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Natural Compounds from Danshen Suppress the Activity of Hepatic Stellate Cells

  • Oh, Seong-Hwan;Cho, Kyung-Hwan;Yang, Beom-Seok;Roh, Yong-Kyun
    • Archives of Pharmacal Research
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    • v.29 no.9
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    • pp.762-767
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    • 2006
  • Danshen is an herbal medication frequently used in oriental medicine to treat liver or kidney malfunction. In the course of our studies, we observed that compounds purified from Danshen exhibit an inhibitory activity against Discoidin Domain Receptor 2 (DDR2) tyrosine kinase. Through this inhibition, these compounds also inhibited the growth of HSC T6 cells and suppressed the expression of alpha-smooth muscle actin and MMP2, as well as collagen synthesis, all of which are increased in activated liver stellate cells. Given that activation of liver stellate cells is the hallmark of liver fibrosis and that DDR2 plays a critical role in this activation, these results suggest that one of the pharmacological activities of Danshen extract that protects the liver is the inhibition of key cell-signaling kinases, such as DDR2, in liver stellate cells.

An improved method to determine hydroxyproline in an immortalized rat liver stellate cell line (HSC-T6)

  • Lee, Eun-Su;Kwak, Wie-Jong;Lee, Eung-Seok;Soh, Yun-Jo
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.154.1-154.1
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    • 2003
  • Hydroxyproline (HYP) is a post-translational product of proline hydroxylation catalyzed by an enzyme prolyl 4-hydroxylase which plays a crucial role in the synthesis of all collagens, because the 4-hydroxyproline residues are essential for the folding of the newly synthesized collagen polypeptide chains into triple-heical molecules. Considering the role of collagen and its significance in many clinically important diseases such as liver cirrhosis, a great deal of attention has been directed toward the development of an assay at cell-based system. (omitted)

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Dimensional Responses of Wood Under Cyclical Changing Temperature at Constant Relative Humidity

  • Yang, Tiantian;Ma, Erni;Shi, Yi
    • Journal of the Korean Wood Science and Technology
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    • v.43 no.5
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    • pp.539-547
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    • 2015
  • To investigate dimensional responses of wood under dynamic temperature condition, poplar (populous euramericana Cv.) specimens, 20 mm in radial (R) and tangential (T) directions with two thicknesses of 4 and 10 mm along the grain, were exposed to cyclic temperature changes in square wave between $25^{\circ}C$ and $40^{\circ}C$ at 60% relative humidity (RH) for three different cycling periods of 6 h, 12 h and 24 h. R and T dimensional changes measured during the cycling gave the following results: 1) Transverse dimensional changes of the specimens were generally square but at an opposite phase and lagged behind the imposed temperature changes. The phase lag was inversely correlated with cycling period, but positively related to specimen thickness, while the response amplitude was directly proportional to cycling period, but in a negative correlation with specimen thickness. 2) The specimens showed swelling hysteresis behavior. The heat shrinkage coefficient (HSC) became greater as cycling period increased or specimen thickness decreased. 3) Dimensional changes of the specimens produced deformation accumulation during repeated adsorption and desorption. The deformation accumulating ratio decreased with an increase in cycling period and specimen thickness. 4) Wood suffered 1.5 times as many dimensional changes per unit temperature variation as per unit humidity variation, and this deformation behaved even more seriously under static condition.

Improved human hematopoietic reconstitution in HepaRG co-transplanted humanized NSG mice

  • Kim, Jin;Ryu, Bokyeong;Kim, Ukjin;Kim, Chang-Hwan;Hur, Gyeung-Haeng;Kim, C-Yoon;Park, Jae-Hak
    • BMB Reports
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    • v.53 no.9
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    • pp.466-471
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    • 2020
  • Several humanized mouse models are being used to study humanspecific immune responses and diseases. However, the pivotal needs of fetal tissues for the humanized mice model have been huddled because of the demand for ethical and medical approval. Thus, we have verified the hematopoietic and immunomodulatory function of HepaRG and developed a new and easy humanized mouse model to replace the use of fetal liver tissue. HepaRG co-transplanted Hu-NSG mice significantly increased CD45+ lymphocytes and CD19+ B cells and CD3+ T cells than normal Hu-NSG, suggesting enhanced reconstitution of the human immune system. These results have improved the applicability of humanized mice by developing new models easily accessible.

MicroRNA-126 Regulates the Expression of Stem Cell Transcription Factors (Sox2 and Lin28) in Various Ovarian Tumors (MicroRNA-126은 난소 종양세포의 줄기세포 전사인자 (Sox2와 Lin28) 발현을 조절한다)

  • Park, Ho;Jekal, Seung Joo
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.4
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    • pp.298-305
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    • 2015
  • Stem cell-like tumor cells are reported to be the main reason for tumor recurrence and metastasis. As one of the new approaches to overcome cancer, studies are emerging to inhibit the expressions of stem cell transcriptional factors (Oct4, Sox2, Klf-4, and Lin28) in cancer cells. MicroRNAs are master genetic regulators that can control development and differentiation of stem cells. In this study using various ovarian tumors (Skov3, Ovcar3, Tov112D, Tov21G, PA-1 and Hsc832(c)T), we examined the expressions of stem cell-related transcription factors, and the biological changes in cell survival and growth by miR-126 that targets stem cell transcriptional factors. We observed that treatment of miR-126 induced the morphological changes and cell suspension in most cells. In addition, miR-126 induced gradual regression of cell division except Skov3 cells, especially significant time-dependent reduction in Tov112D, Tov21G and PA-1. When we examined the expression of stem cell transcriptional factors, Sox2 was shown to be down-regulated after miR-126. Our results demonstrate that miR-126 treatment can provide the reversible environment to regulate cell division and to induce cell death of ovarian tumors, suggesting the molecular biological clues for clinical usage.