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

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Anticancer activity of subfractions containing pure compounds of Chaga mushroom (Inonotus obliquus) extract in human cancer cells and in Balbc/c mice bearing Sarcoma-180 cells

  • Chung, Mi-Ja;Chung, Cha-Kwon;Jeong, Yoon-Hwa;Ham, Seung-Shi
    • Nutrition Research and Practice
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    • v.4 no.3
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    • pp.177-182
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    • 2010
  • The Chaga mushroom (Inonotus obliquus) has been used in folk medicine to treat cancers. However, limited information exists on the underlying anticancer effects of the major component of I. obliquus in vivo. We hypothesize that the pure compounds ($3{\beta}$-hydroxy-lanosta-8,24-dien-21-al, inotodiol and lanosterol, respectively) separated from I. obliquus would inhibit tumor growth in Balbc/c mice bearing Sarcoma-180 cells (S-180) in vivo and growth of human carcinoma cells in vitro. To test this hypothesis, the growth inhibition of each subfraction isolated from I. obliquus on human carcinoma cell lines (lung carcinoma A-549 cells, stomach adenocarcinoma AGS cells, breast adenocarcinoma MCF-7 cells, and cervical adenocarcinoma HeLa cells) was tested in vitro. Then, after S-180 implantation, the mice were fed a normal chow supplemented with 0, 0.1 or 0.2 mg of subfraction 1, 2 or 3 per mouse per day. All of the subfractions isolated from I. obliquus showed significant cytotoxic activity against the selected cancer cell lines in vitro. Subfraction 1 was more active than subfraction 2 and subfraction 3 against the A549, AGS and MCF-7 cancer cell lines in vitro. In in vivo results, subfraction 1 isolated from I. obliquus at concentrations of 0.1 and 0.2 mg/mouse per day significantly decreased tumor volume by 23.96% and 33.71%, respectively, as compared with the control. Subfractions 2 and 3 also significantly inhibited tumor growth in mice bearing S-180 as compared with the control mouse tumor. Subfraction 1 isolated from I. obliquus showed greater inhibition of tumor growth than subfractions 2 and 3, which agrees well with the in vitro results. The results suggest that I. obliquus and its compounds in these subfractions isolated from I. obliquus could be used as natural anticancer ingredients in the food and/or pharmaceutical industry.

Ginsenoside Rh2 inhibiting HCT116 colon cancer cell proliferation through blocking PDZ-binding kinase/T-LAK cell-originated protein kinase

  • Yang, Jianjun;Yuan, Donghong;Xing, Tongchao;Su, Hongli;Zhang, Shengjun;Wen, Jiansheng;Bai, Qiqiang;Dang, Dongmei
    • Journal of Ginseng Research
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    • v.40 no.4
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    • pp.400-408
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    • 2016
  • Background: Ginsenoside Rh2 (GRh2) is the main bioactive component in American ginseng, a commonly used herb, and its antitumor activity had been studied in previous studies. PDZ-binding kinase/T-LAK cell-originated protein kinase (PBK/TOPK), a serine/threonine protein kinase, is highly expressed in HCT116 colorectal cancer cells. Methods: We examined the effect of GRh2 on HCT116 cells ex vivo. Next, we performed in vitro binding assay and in vitro kinase assay to search for the target of GRh2. Furthermore, we elucidated the underlying molecular mechanisms for the antitumor effect of GRh2 ex vivo and in vivo. Results: The results of our in vitro studies indicated that GRh2 can directly bind with PBK/TOPK and GRh2 also can directly inhibit PBK/TOPK activity. Ex vivo studies showed that GRh2 significantly induced cell death in HCT116 colorectal cancer cells. Further mechanistic study demonstrated that these compounds inhibited the phosphorylation levels of the extracellular regulated protein kinases 1/2 (ERK1/2) and (H3) in HCT116 colorectal cancer cells. In vivo studies showed GRh2 inhibited the growth of xenograft tumors of HCT116 cells and inhibited the phosphorylation levels of the extracellular regulated protein kinases 1/2 and histone H3. Conclusion: The results indicate that GRh2 exerts promising antitumor effect that is specific to human HCT116 colorectal cancer cells through inhibiting the activity of PBK/TOPK.

Effects of Samultang on Glutamate-Induced Apoptosis of Hippocampus Cells (사물탕(四物湯)이 Glutamate에 의한 해마세포의 손상에 미치는 영향)

  • Jeong, Dae-Young;Choi, Chul-Won;Moon, Byung-Soon
    • The Journal of Korean Medicine
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    • v.30 no.1
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    • pp.64-75
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    • 2009
  • Objective: This study was designed to investigate the effect of Samultang (SMT) under hippocampus cells ischemia both in vitro and in vivo. Methods: In the in vitro study, HT22 cells, predominantly detected in the cytoplasm, which coincides with the location of the mitochondria, were used as indicators. In the in vivo study, permanent middle cerebral artery occlusion (MCAO) was induced on rats. SMT was given orally 2 h before induction of permanent focal brain ischemic injury. Result: In the in vitro study, SMT had protective effects in glutamate-induced cytotoxicity, which was revealed as apoptosis characterized by chromatic condensation and the loss of mitochondrial membrane potential in HT22 cells. In the in vivo study, TTC (2,3,5-triphenyltetrazolium chloride) staining showed a marked ischemic injury in blood supply territory of the middle cerebral artery (MCA) such as the cerebral cortex and striatum. However, treatment with SMT significantly reduced infarcted volume. SMT increased marked survival of HT22 cells against glutamate-induced cytotoxicity in MTT assay. Conclusion: These results suggest that water extract of SMT provides neuroprotection against ischemic or oxidative injury by inhibition of apoptotic cell death.

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Three-step in vitro digestion model for evaluating and predicting fecal odor emission from growing pigs with different dietary protein intakes

  • Lo, Shih-Hua;Chen, Ching-Yi;Wang, Han-Tsung
    • Animal Bioscience
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    • v.35 no.10
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    • pp.1592-1605
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    • 2022
  • Objective: The objective of this study was to select an effective in vitro digestion-fermentation model to estimate the effect of decreasing dietary crude protein (CP) on odor emission during pig production and to suggest potential prediction markers through in vitro and in vivo experiments. Methods: In the in vitro experiment, three diet formulations with different CP contents (170 g/kg, 150 g/kg, and 130 g/kg) but containing the same standardized ileal digestible essential amino acids (SID-EAA) were assessed. Each diet was evaluated by two different in vitro gastric-intestinal phase digestion methods (flask and dialysis), combined with fresh pig feces-ferment inoculation. Eighteen growing barrows (31.9±1.6 kg) were divided into three groups: control diet (180 g CP/kg, without SID-EAA adjustment), 170 g CP/kg diet, and 150 g CP/kg diet for 4 weeks. Results: The in vitro digestion results indicated that in vitro digestibility was affected by the gastric-intestinal phase digestion method and dietary CP level. According to the gas kinetic and digestibility results, the dialysis method showed greater distinguishability for dietary CP level adjustment. Nitrogen-related odor compounds (NH3-N, indole, p-cresol, and skatole) were highly correlated with urease and protease activity. The feeding study indicated that both EAA-adjusted diets resulted in a lower odor emission especially in p-cresol and skatole. Both protease and urease activity in feces were also closely related to odor emissions from nitrogen metabolism compounds. Conclusion: Dialysis digestion in the gastric-intestinal phase followed by fresh fecal inoculation fermentation is suitable for in vitro diet evaluation. The enzyme activity in the fermentation and the fecal samples might provide a simple and effective estimation tool for nitrogen-related odor emission prediction in both in vitro and in vivo experiments.

Enhancement of Chromosome Aberrations in Lymphocytes of Mice after in Vivo Exposure to Chemicals and in Vitro Challenge with Bleomycin (MNNG 또는 Benzo(a)pyrene 유도 염색체 이상에 미치는 Bleomycin의 효과)

  • Heo, M.Y.;Grady, J.J.;Au, W.W.
    • Environmental Mutagens and Carcinogens
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    • v.18 no.2
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    • pp.71-76
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    • 1998
  • Exposure to environmental toxicants can cause cellular problems including the interference of DNA repair processes which may lead to the development of cancer. The existence of toxicant-induced DNA repair abnormality was investigated using mice exposed in vivo to genotoxic chemicals and then challenging their exposed lymphocytes in vitro with bleomycin. The repair of bleomycin-induced DNA damage as estimated by the frequency of chromosome aberrations was determined. Our data indicates that the observed aberration frequencies after in vivo exposure to N-methyl-N'-nitro-N-nitnsoguanidine (MNNG) and in vitro challenge with bleomycin are consistently higher than expected. The enhanced response is not due to the induction of chromosome damage by 25 or 50 mg/kg MNNG since the chemical did not cause chromosome aberrations in lymphocytes of these mice. The observed response after the combined exposure to benzo[a]pyrene (BP) and bleomycin was significantly lower than expected with low in vivo doses of BP (50 mg/kg) and then significantly higher than expected with the high doses (200 mg/kg). We interpret our data to indicate that in vivo exposure to genotoxic agents can cause abnormal DNA repair activities. The response is, however, independent of the clastogenic activities of the inducing chemicals, but dependent upon the inducing agents and on the exposure doses.

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Efficacy Evaluation of Tissue Inhibitor of Metalloproteinases-2 and Endostatin on Angiogenesis (Tissue Inhibitor of Metalloproteinases-2와 Endostatin의 혈관신생 제어 효능 평가)

  • Kim, Soo-Hyeon;Cho, Young-Rak;Yoon, Hyun-Jae;Ko, Hee-Young;Kim, Pyeung-Hyeun;Seo, Dong-Wan
    • YAKHAK HOEJI
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    • v.54 no.6
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    • pp.488-493
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    • 2010
  • Therapeutic manipulation of angiogenesis, the formation of new vascular sprouts from existing capillaries, is one of the promising strategies for treatment of human diseases such as cancer, arthritis, and cardiovascular disorder. In the present study, we examined the effects and molecular mechanism of tissue inhibitor of metalloproteinases-2 (TIMP-2) and endostatin on fibroblast growth factor-2 (FGF-2)-stimulated endothelial cell proliferation, migration and adhesion in vitro, and angiogenesis in vivo. TIMP-2 and endostatin showed potent anti-angiogenic activity in vitro and in vivo. These effects appear to be mediated through different angiogenic signaling pathways. Collectively, our findings demonstrate that TIMP-2 and endostatin strongly inhibit FGF-2-induced angiogenic responses, and the establishment of fast and reproducible evaluation system in vitro and in vivo for the development of anti-angiogenic biomaterials and therapeutics.

In Vitro and in Vivo Antifungal Activities of 6-[(N-4-bromophenyl)amino]-7-chloro-5,8-quinolinediones

  • Ryu, Chung-Kyu;Kim, Dong-Hyun;Kwon, Sang-Mee;Jung, Sung-Hee;Kim, Sung-Hee
    • Archives of Pharmacal Research
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    • v.20 no.6
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    • pp.586-589
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    • 1997
  • Antifungal activities of 6-[(N-4-bromophenyl)amino]-7-chloro-5,8-quinolinedione (RCK7) were tested. The MIC values of RCK7 were determined for antifungal suceptibility, in vitro against Aspergillus niger, Cryptococcus neoformans and Trichophyton mentagrophyte by standard agar streak method. In vitro, RCK7 showed more potent antifungal activity than fluconazole and ketoconazole. Also, RCK7 was tested for in vivo antifungal activity in the treatment of systemic infection with Candida albicans in normal mice. The therapeutic potential of RCK7 had been assessed by evaluating their survival rate against systemic infections compared with that of ketoconazole. $ED_{50}$ of intraperitoneally administered RCK7 ws $2.05{\pm}0.30mg/kg$ but that of ketoconazole was $8.00{\pm}0.73 mg/kg$, respectively. When RCK7 was administered intravenously at the $ED_{50}$(2.05 mg/kg). the colony counts of Candida albicans in the liver after 7 days and 14 days were reduced as likely as ketoconazole at the $ED_{50}(8.00 mg/kg)$, and the better survival rates than ketoconazole's were achieved after 14 days. The results suggest that RCK7 may be a potent antifungal agent.

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Comparison of the Digestibility of Grain and Forage by Sheep, Red and Fallow Deer

  • Ru, Y.J.;Glatz, P.C.;Miao, Z.H.;Swanson, K.;Falkenberg, S.;Wyatt, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.6
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    • pp.800-805
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    • 2002
  • Two experiments were conducted to compare digestibility of 12 diets in sheep, red and fallow deer. No differences (p>0.05) between sheep, red and fallow deer in digestibility of dry matter, organic matter and digestible energy content for all diets were found except for the sorghum diet and medic hay. Sheep and fallow deer digested the sorghum diet better than red deer. An in vitro study showed that sheep had a lower in vitro dry matter digestibility and digestible energy content than both red and fallow deer, with a significant interaction between animal species and feed ingredient. Deer digested straws and hays better (p<0.05) than sheep. In vitro digestibility was lower (p<0.05) than in vivo digestibility, but significantly correlated with in vivo digestibility for red and fallow deer. The in vitro method for digestibility estimation has potential as a rapid feed evaluation system for deer, but needs further validation.

In vitro and In vivo Activities of a Biocontrol Agent, Serratia plymuthica A2l-4, Against Phytophthora capsici

  • Shen, Shun-Shan;Park, Ok-Hee;Lee, Sun-Mi;Park, Chang-Seuk
    • The Plant Pathology Journal
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    • v.18 no.4
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    • pp.221-224
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    • 2002
  • In vitro and in vivo activities of a biocontrol agent, Serratia plymuthica strain A2l-4, was evaluated for the control of Phytophthora blight of pepper, Strain A2l-4 inhibited mycelial growth, germination of zoosporangia and cystospores, and formation of zoospore and zoosporangia of Phytophthora capsici in vitro. In the pot experiment, incidence of Phytophthora blight of pepper in non-treated control was 100% at 14 days after inoculation, while no disease was observed in the plot treated with S. plymuthica A2l-4. In the greenhouse test, infection rate of pepper in the non-treated plots was 74.5%, while it was only 12.6% in the plots treated with A2l-4. Results indicate that S. plymuthica A2l-4 is a potential biocontrol agent for Phytophthora blight of pepper.

Screening and Characterization of Lactic Acid Bacteria Strains with Anti-inflammatory Activities through in vitro and Caenorhabditis elegans Model Testing

  • Lee, Hye Kyoung;Choi, Sun-Hae;Lee, Cho Rong;Lee, Sun Hee;Park, Mi Ri;Kim, Younghoon;Lee, Myung-Ki;Kim, Geun-Bae
    • Food Science of Animal Resources
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    • v.35 no.1
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    • pp.91-100
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    • 2015
  • The present study was conducted to screen candidate probiotic strains for anti-inflammatory activity. Initially, a nitric oxide (NO) assay was used to test selected candidate probiotic strains for anti-inflammatory activity in cultures of the murine macrophage cell line, RAW 264.7. Then, the in vitro probiotic properties of the strains, including bile tolerance, acid resistance, and growth in skim milk media, were investigated. We also performed an in vitro hydrophobicity test and an intestinal adhesion assay using Caenorhabditis elegans as a surrogate in vivo model. From our screening, we obtained 4 probiotic candidate lactic acid bacteria (LAB) strains based on their anti-inflammatory activity in lipopolysaccharide (LPS)-stimulated RAW 264.7 cell cultures and the results of the in vitro and in vivo probiotic property assessments. Molecular characterization using 16S rDNA sequencing analysis identified the 4 LAB strains as Lactobacillus plantarum. The selected L. plantarum strains (CAU1054, CAU1055, CAU1064, and CAU1106) were found to possess desirable in vitro and in vivo probiotic properties, and these strains are good candidates for further investigations in animal models and human clinical studies to elucidate the mechanisms underlying their anti-inflammatory activities.