• Title/Summary/Keyword: in vitro/in vivo studies

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Modulacon of Cell Cycle Control by Histone Deacetylase Inhibitor Trichostatin A in A549 Human Non-small Cell Lung Cancer Cells (인체폐암세포 A549의 세포주기 조절인자에 미치는 histone deacetylase inhibitor trichostatin A의 영향)

  • Hwang Ji Won;Kim Young Min;Hong Su Hyun;Choi Byung Tae;Lee Won Ho;Choi Yung Hyun
    • Journal of Life Science
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    • v.15 no.5 s.72
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    • pp.726-733
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    • 2005
  • Histone deacetylase (HDAC) inhibitors target key steps of tumor development. They inhibit proliferation, induce differentiation and/or apoptotic cell death, and exhibit potent antimetastatic and antiangiogenic properties in cancer cells in vitro and in vivo. Although they are emerging as a promising new treatment strategy in malignancy, how they exert their effect on human non-small cell lung cancer cells is as yet unclear. The present study was undertaken to investiate the underlying mechanism of a HDAC inhibitor trichostatin A (TSA)-induced growth arrest and its effect on the cell cycle control gene products in a human lung carcinoma cell line A549. TSA treaoent induced the growth inhibition and morphological changes in a concentration-dependent manner. Treatment of A549 cells with TSA resulted in a concentration-dependent increased G1 (under 100 ng/ml) and/or G2/M (200 ng/ml) cell population of the cell cycle as determined by flow cytometry Moreover, 200 ng/ml TSA treatment significantly induced the population of sub-G1 cells (23.0 fold of control). This anti-proliferative effect of TSA was accompanied by a marked inhibition of cyclins, positive regulators of cell cycle progression, and cyclin-dependent kinases (Cdks) expression and concomitant induction of tumor suppressor p53 and Cdk inhibitors such as p21 and p27 Although further studies are needed, these findings provide important insights into the possible molecular mechanisms of the anti-cancer activity of TSA in human lung carcinoma cells.

Anti-inflammatory activities of Sagunja-tang(Sijunzi-tang) (사군자탕(四君子湯)의 항염증 효능 연구)

  • Lee, Jin-Ah;Ha, Hye-Kyung;Jung, Da-Young;Lee, Ho-Young;Lee, Nam-Hun;Lee, Jun-Kyoung;Seo, Chang-Seob;Kim, Jung-Hoon;Lee, Mee-Young;Shin, Hyeun-Kyoo
    • Herbal Formula Science
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    • v.18 no.1
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    • pp.87-94
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    • 2010
  • Objectives : To provide the information of efficacy for Sagunja-tang (Sijunzi-tang; SG), it was evaluated the anti-inflammatory effect. SG, a widely used herbal formula in tranditional Korean medicine, has been used to treat for the Boki-invigorating. In many studies, plant-derived anti-inflammatory efficacies have been investigated for their potential inhibitory effects on lipopolysaccharide (LPS)-stimulated macrophages. This study was performed to examine the anti-inflammatory effects of SG extract on LPS-stimulated RAW 264.7 cells. Methods : The productions of nitric oxide (NO), prostaglandin (PG)$E_2$, interleukin (IL)-6 and tumor necrosis factor (TNF)-$\alpha$ were examined in a macrophage cell line, RAW 264.7 cells, in the presence of the SG extract. RAW 264.7 cells were incubated with LPS $1\;{\mu}g/mL$ and SG extract for 18 hours. The anti-inflammatory activity of SG was investigated by carrageenin-induced paw edema in rats. The paw volume was measured at 0, 2 and 4 hours following carrageenin-induced paw edema in rats. Results : SG extract showed inhibitory effect on $PGE_2$, IL-6 and TNF-$\alpha$ by LPS-stimulated RAW 264.7 cells. But SG extract was not inhibitory effect on NO by LPS-stimulated RAW 264.7 cells. And administration of SG extract (1 g/kg) showed a reduction in carrageenin-induced paw edema on rats. Conclusions : These results suggest that SG extract has anti-inflammatory activities in vitro and in vivo models.

Effects of β-Glucan from Paenibacillus polymyxa and L-theanine on Growth Performance and Immunomodulation in Weanling Piglets

  • Hwang, Y.H.;Park, B.K.;Lim, J.H.;Kim, M.S.;Song, I.B.;Park, S.C.;Jung, H.K.;Hong, J.H.;Yun, H.I.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.12
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    • pp.1753-1759
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    • 2008
  • Forty weanling piglets ($5.6{\pm}0.5kg$ and 26 to 30 d of age) were used in a 28-d experiment to determine the effects of ${\beta}$-glucan from Paenibacillus polymyxa and L-theanine on growth performance. Piglets were randomly allotted to four groups (n = 10, 2 animals per pen) provided with the basal feed (control), ${\beta}$-glucan 400 mg/kg feed, L-theanine 80 mg/kg feed or ${\beta}$-glucan plus l-theanine (combination of the above-mentioned concentrations). Body weight and feed consumption were recorded during four weeks. Subsequently, the immunomodulatory effects of ${\beta}$-glucan and L-theanine were investigated for lipopolysaccharide (LPS)-induced cytokine production in vitro and in vivo on day 28. Although there were no significant differences in the growth performances among the treatment groups, ${\beta}$-glucan plus L-theanine had 5.6% greater ADG (p = 0.074) on day 21 to 28. ${\beta}$-Glucan alone or plus L-theanine increased interleukin (IL)-10 levels and decreased interferon (IFN)-$\gamma$ and tumor necrosis factor (TNF)-${\alpha}$ levels in cultured medium by LPS treatment (p<0.05). Plasma IL-10 levels were also increased in the piglets fed with ${\beta}$-glucan alone or plus L-theanine after LPS challenge ($25{\mu}g/kg$, i.p.), whereas plasma IFN-$\gamma$ and TNF-${\alpha}$ levels were decreased (p<0.05). The levels of IFN$\gamma$ in piglets fed with ${\beta}$-glucan plus L-theanine showed the greatest inhibition after LPS challenges. In conclusion, treatment of ${\beta}$-glucan alone or plus L-theanine might lessen inflammatory responses against Gram-negative bacterial infection via the inhibition of pro-inflammatory cytokine production and enhancement of anti-inflammatory cytokine production. Further studies are needed to determine an optimal concentration of ${\beta}$-glucan and L-theanine for improved growth performance.

Polyphenol Contents and Antioxidant Activities of Lentil Extracts from Different Cultivars (품종별 렌틸 추출물의 폴리페놀화합물 함량 및 항산화 활성)

  • Lee, So-Hee;Lee, Syng-Ook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.7
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    • pp.973-979
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    • 2016
  • Lentils (Lens culinaris) have been gaining increasing attention recently as a top five superfood, as they are high in protein and other essential nutrients, including folate, iron, potassium, and various antioxidants. In the present study, phenolic extracts from four different lentil cultivars (green, red, French, and beluga) were evaluated for their total phenolic contents and in vitro antioxidant activities. Total polyphenol and flavonoid contents of four different lentil extracts were 27.30~30.30 mg tannic acid equivalents (TAE)/g and 13.14~16.29 mg quercetin equivalents (QUE)/g, respectively. Beluga and red lentil extracts showed higher polyphenol contents than others (P<0.05), whereas there was no significant difference in flavonoid contents among the four lentil cultivars. $RC_{50}$ values of the lentil extracts for DPPH radical, ABTS radical, and $H_2O_2$ were $57.42{\sim}64.49{\mu}g/mL$, $66.11{\sim}75.69{\mu}g/mL$, and $59.72{\sim}72.86{\mu}g/mL$, respectively. Among the four lentil extracts, beluga lentil extract showed the most potent scavenging effect in all three reactive oxygen species (ROS) scavenging assays, and thus beluga extract was further tested for its inhibitory effect on early peroxidation of linoleic acid. The results showed that beluga lentil extract significantly inhibited linoleic acid peroxidation in a dose-dependent manner (concentration required for 50% reduction=$222.76{\m}g/mL$). In addition, beluga lentil extract showed a significant protective effect against alcohol-induced cytotoxicity in AML-12 cells (normal mouse hepatocyte cell line). Taken together, these results suggest that lentil extracts represent potential sources of natural antioxidants, and further studies will be necessary to determine their protective effects against oxidative stress in vivo.

Effect of low frequency oscillations during milking on udder temperature and welfare of dairy cows

  • Antanas Sederevicius;Vaidas Oberauskas;Rasa Zelvyte;Judita Zymantiene;Kristina Musayeva;Juozas Zemaitis;Vytautas Jurenas;Algimantas Bubulis;Joris Vezys
    • Journal of Animal Science and Technology
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    • v.65 no.1
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    • pp.244-257
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    • 2023
  • The study aimed to investigate the effect of low-frequency oscillations on the cow udder, milk parameters, and animal welfare during the automated milking process. The study's objective was to investigate the impact of low-frequency oscillations on the udder and teats' blood circulation by creating a mathematical model of mammary glands, using milkers and vibrators to analyze the theoretical dynamics of oscillations. The mechanical vibration device developed and tested in the study was mounted on a DeLaval automatic milking machine, which excited the udder with low-frequency oscillations, allowing the analysis of input parameters (temperature, oscillation amplitude) and using feedback data, changing the device parameters such as vibration frequency and duration. The experimental study was performed using an artificial cow's udder model with and without milk and a DeLaval milking machine, exciting the model with low-frequency harmonic oscillations (frequency range 15-60 Hz, vibration amplitude 2-5 mm). The investigation in vitro applying low-frequency of the vibration system's first-order frequencies in lateral (X) direction showed the low-frequency values of 23.5-26.5 Hz (effective frequency of the simulation analysis was 25.0 Hz). The tested values of the first-order frequency of the vibration system in the vertical (Y) direction were 37.5-41.5 Hz (effective frequency of the simulation analysis was 41.0 Hz), with higher amplitude and lower vibration damping. During in vivo experiments, while milking, the vibrator was inducing mechanical milking-similar vibrations in the udder. The vibrations were spreading to the entire udder and caused physiotherapeutic effects such as activated physiological processes and increased udder base temperature by 0.57℃ (p < 0.001), thus increasing blood flow in the udder. Used low-frequency vibrations did not significantly affect milk yield, milk composition, milk quality indicators, and animal welfare. The investigation results showed that applying low-frequency vibration on a cow udder during automatic milking is a non-invasive, efficient method to stimulate blood circulation in the udder and improve teat and udder health without changing milk quality and production. Further studies will be carried out in the following research phase on clinical and subclinical mastitis cows.

Measurement of CYP450 Enzymes Activity of Bosentan in HepaRG Cell (HepaRG 세포를 이용한 Bosentan 약물의 CYP450 효소활성 측정)

  • Han, Kyoung-Moon;Jung, Jung-A;Sin, Ji-Soon;Cha, Hye-Jin;Bae, Young-Ji;Kim, Hyun-Uk;Kim, Young-Hoon;Seong, Won-Keun;Kang, Hoil
    • YAKHAK HOEJI
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    • v.58 no.4
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    • pp.255-261
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    • 2014
  • Poly-pharmacy has been on the rise because of aging of population and chronic disease. Most of drug metabolism happens in the liver by CYP isozymes and the metabolism by CYP450 enzymes. The Cytochrome P450 (CYP) is a superfamily of enzymes that catalyzes the oxidations of many endogenous and exogenous compounds. Primary human Hepatocytes (HH) are considered as the gold standard model for In vitro drug interaction studies. However, there are several limitations (cost, limited life span) for using HH cells. HepaRG cells are being used as a possible alternative. HepaRG cells were cultured in William E medium containing the positive control inducers (1A2: 10, 25, 50 ${\mu}M$ omeprazole, 2C9 and 2C19: 10 ${\mu}M$ rifampin, 3A4: 10, 25, 50 ${\mu}M$ rifampin) at $37^{\circ}C$, 5 % $CO_2$ in a humidified atmosphere. This study was to evaluate the induction of CYP isozymes (1A2, 2C9, 2C19 and 3A4) using LC-MS/MS. We evaluated the potential induction ability of Bosentan, as a drug of pulmonary artery hypertension, in HepaRG cells. For reference, dose of the Bosentan is determined to the basis of the $C_{max}$ (835 mg/ml) value. The enzyme activity demonstrated that CYP2C9 and 3A4 were induced up to 20 times by Bosentan. Like as In vivo, the enzyme activity of CYP2C9 and CYP3A4 is significantly induced in a dose-dependent by Bosentan.

CK2 phosphorylates AP-2α and increases its transcriptional activity

  • Ren, Kaiqun;Xiang, Shuanglin;He, Fangli;Zhang, Wenfeng;Ding, Xiaofeng;Wu, Yanyang;Yang, Liping;Zhou, Jianlin;Gao, Xiang;Zhang, Jian
    • BMB Reports
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    • v.44 no.7
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    • pp.490-495
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    • 2011
  • Transcription factor AP-$2{\alpha}$ involves in the process of mammalian embryonic development and tumorigenesis. Many studies have shown that AP-$2{\alpha}$ functions in association with other interacting proteins. In a two-hybrid screening, the regulatory subunit ${\beta}$ of protein casein kinase 2 ($CK2{\beta}$) was identified as an interacting protein of AP-$2{\alpha}$; we confirmed this interaction using in-vitro GST pull-down and in-vivo co-immunoprecipitation assays; in an endogenous co-immunoprecipitation experiment, we further found the catalytic subunit ${\alpha}$ of protein casein kinase 2 ($CK2{\alpha}$) also exists in the complex. Phosphorylation analysis revealed that AP-$2{\alpha}$ was phosphorylated by CK2 kinase majorly at the site of Ser429, and such phosphorylation could be blocked by CK2 specific inhibitor 4,5,6,7-tetrabromobenzotriazole (TBB) in a dose-dependent manner. Luciferase assays demonstrated that both $CK2{\alpha}$ and $CK2{\beta}$ enhanced the transcription activity of AP-$2{\alpha}$; moreover, $CK2{\beta}$ increased the stability of AP-$2{\alpha}$. Our data suggest a novel cellular function of CK-2 as a transcriptional co-activator of AP-$2{\alpha}$.

In Vitro Culture of Human Nasal Epithelial Cells by Monolayer Culture of Dissociated Cells (분리 세포의 단층세포 배양법에 의한 인체 비점막 상피세포의 배양)

  • Kim, Yong-Dae;Song, Si-Youn;Min, Myung-Ki;Sub, Jang-Su;Song, Kei-Won;Park, Ho-Sun
    • Journal of Yeungnam Medical Science
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    • v.15 no.2
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    • pp.286-296
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    • 1998
  • Different techniques for culturing respiratory epithelial cells have been developed to overcome the limitations of studies on in vivo and on bioptic material. Traditionally, culture systems are divided into organ cultures, explant cultures and dissociated cell cultures. The first two contain both epithelial and non-epithelial cells. However, in monolayer cultures of dissociated cells only epithelial cells are present, the effects observed are caused by a pure epithelial responses. The purpose of this study is to establish primary culture method of human nasal epithelium (HNEC) by monolayer culture of dissociated cells to evaluate the role of the epithelial cells in the allergic and non-allergic nasal inflammatory reactions. HNEC was prepared by primary culture method of monolayer culture of dissociated cells from human inferior nasal turbinate mucosa of septal deviation patients. Primary cultured cells were characterized by indirect immunofluorescence assay and transmission electron microscopy. The immunoreactivities of cytokeratin-pan and cytokeratin No. 8 were observed in cultured HNEC. However, the immnoreactivities of vimentin and von Willebrand factor were not observed in cultured HNEC. The tonofilaments and desmosome were observed in cultured HNEC. The cultured epithelial cells were identified to be pure nasal epithelial cells. The monolayer culture of dissociated cells could successfully be employed for further study to investigate the role of the epithelial cells in allergic or non-allergic nasal inflammatory diseases.

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Inhibitory Effects of a New Herbal Composition (HemoHIM) on UVB-Induced Suppression of Langerhans Cell's Accessory Cell Function (생약복합조성물(HemoHIM)의 자외선 조사로 억제된 랑게르한스 세포의 항원제시기능 방호효과)

  • Kim, Jong-Jin;Jo, Sung-Kee;Jung, U-Hee;Park, Hae-Ran;Yee, Sung-Tae
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1761-1771
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    • 2011
  • In the previous results, we developed an effective products to apply as functional foods for overcome of radiation damage and reduction of side effects in radiotherapy. To verify the prevention of UVB-induced immunosuppression of immune cell function by HemoHIM, we studied on the mechanism of the skin immune function for the protection in UVB irradiation. In studies presented here, we showed that HemoHIM can prevent UVB-induced impairment of skin immune cell function by in vitro and in vivo assay. Exposure of freshly cultured murine dendritic cells (DCs) with IL-4/GM-CSF to UVB irradiation resulted in impairment of accessory function. This suppression could be prevented by addition of HemoHIM before or after to the cultures of UVB-irradiated DCs. We also tested the effects of HemoHIM on the suppression of contact hypersensitivity (CHS) treated oral or intraperitoneal administration. This UVB-suppressed CHS was prevented by administration of HemoHIM to UVB-irradiated mice. These results suggest that HemoHIM may prevent UVB-induced immune suppression in the skin.

Anti-cancer effects of kelp extract in mouse melanoma B16-F0 cell line through apoptosis (마우스 흑색종 세포주 B16-F0에서 다시마 추출물의 세포사멸을 통한 항암 효과)

  • Lee, Seong-Uk;Kim, Yoon Hee
    • Korean Journal of Food Science and Technology
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    • v.54 no.2
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    • pp.134-140
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    • 2022
  • Kelp belongs to the brown algae family and has been reported to exert anti-cancer effects on some cancer types, however studies have not been reported on the anti-cancer effects of kelp extracts on melanoma. In this study, the anti-cancer effects of kelp extract in B16-F0 cells were investigated, and the underlying molecular mechanisms were assessed. Kelp extract was found to inhibit the proliferation of B16-F0 cells, induce cytotoxicity, inhibit cell colony formation, and induce DNA fragmentation and apoptosis. The molecular mechanism was found to involve kelp extract increasing the expression of cytochrome-c and activated caspase-9 in the intrinsic apoptotic pathway. In addition, kelp extract upregulated the expression of Fas-associated protein with death domain and activated caspase-8 in the extrinsic apoptosis pathway. Activation of caspase-9 and caspase-8 by kelp extract induced activation of caspase-3 and cleaved poly adenosine diphosphate-ribose polymerase, consequently inducing apoptosis. These data suggest that kelp extract represents a potential therapeutic agent for melanoma.