• 제목/요약/키워드: in vivo tissue

검색결과 865건 처리시간 0.028초

The Effect of L-Ornithine on the Phosphorylation of mTORC1 Downstream Targets in Rat Liver

  • Kokubo, Takeshi;Maeda, Shyuichi;Tazumi, Kyoko;Nozawa, Hajime;Miura, Yutaka;Kirisako, Takayoshi
    • Preventive Nutrition and Food Science
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    • 제20권4호
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    • pp.238-245
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    • 2015
  • A non-protein amino acid, L-ornithine (Orn), has been shown to stimulate the urea cycle and tissue protein synthesis in the liver. The purpose of the current study was to assess whether Orn affects the mammalian target of rapamycin (mTOR) complex 1 (mTORC1) pathway, which is involved in protein synthesis. Primary cultured cells isolated from Wistar rat liver were incubated in an amino acid-free medium, followed by addition of Orn for 3 h. The cell lysate was subjected to immunoblotting to evaluate the phosphorylation of downstream targets of mTORC1, including p70S6K, S6, and 4EBP1. To assess the involvement of mTORC1 for the effect of Orn, the cells were pretreated with the mTOR inhibitor rapamycin before the addition of Orn and the cell lysate was subjected to immunoblotting. We next examined whether the effects of Orn were exerted in vivo. Orn was orally administered to 18 h food-deprived rats, the blood and the livers were collected at 1 and 3 h after administration for immunoblotting. Orn treatment for primary cultured cells for 3 h enhanced the phosphorylation of p70S6K, S6, and 4EBP1. In addition, rapamycin blocked the effects of Orn completely (p70S6K and S6) or partially (4EBP1). The oral administration of Orn to the rat also augmented the phosphorylation of mTORC1 downstream targets notably in S6 at 1 h. Our findings demonstrate that Orn has the potential to induce the phosphorylation of downstream targets of mTORC1 in the rat liver. This may be mediated by the augmentation of mTORC1 activity.

방풍통성산가미방(防風通聖散加味方)이 비만유도(肥滿誘導) 백풍(白嵐)와 지방세포(脂肪細胞) 분화(分化)에 미치는 영향(影響) (Anti-obestic Effects of Bangpoongtongsungcankamibang(BTSK) in 3%3-L1 preadipocyte cells and the Lipid Metabolism of Obesity induced Rats)

  • 이진용;김덕곤;안홍식
    • 대한한방소아과학회지
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    • 제20권1호
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    • pp.219-240
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    • 2006
  • Objective : This experimental study was designed to determine the anti-obestic effects of Bangpoongtongsungcankamibang(BTSK). Method : In vitro, BTSK extracts of various concentration (50,100, 200 $200{\mu}g/ml$)were added examination. The protein and mRNA expression of $C/EBP{\alpha}$ and $PPAR{\gamma}$ receptor was measured by western blot assay and RT-PCR. In vivo, BTSK extracts of various concentration (100, $200{\mu}g/ml$) were orally administered to induced hyperlipidemic rats by poloxamer-407 for consecutive four weeks and serum triglyceride, total cholesterol were measured. This method applied to induced hyperlipidemic rats by triton WR-1339, too. Obesity induced rats by the high fat-diet for six weeks were orally administered BTSK extracts of various concentration (100, $200{\mu}g/ml$) and serum triglyceride, total cholesterol, LDL-cholesterol, triglyceride, LDL-cholesterol, triglyceride, HDL-cholesterol, hydroxy radical, superoxide dismuatse activity were measured. Results : I. In vitro 1. The 3T3-L1 cells' differentiation was significantly decreased by BTSK. 2. expression of $C/EBP{\alpha}$ and $PPAR{\gamma}$ was was significantly decreased by BTSK. II. In vitro 1. BTSK significantly reduced serum triglyceride, total cholesterol, contents in poloxamer-407 treated rat. 2. BTSK significantly reduced serum triglyceride, contents in Triton WR-1339 treated rat. But Total cholesterol did not show a significant change. 3. BTSK significantly reduced body weight gain of rat and adipose tissue mass of rats and serum triglyceride, LDL-cholesterol, contents and significantly increased HDL-cholesterol, HTR(HDL-cholesterol/Total-cholesterol) in rats with obesity induced by the high fat-diet. 4. BTSK reduced blood lipid peroxide, hydroxy radical and increased superoxied dismuatse(SOD) activity.

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제브라피쉬(Danio rerio)를 이용한 비소 고함류 3종 해조류 추출물의 급성 독성평가 (Acute Toxicity Assessment in Zebrafish Danio rerio of Arsenic-rich Extracts from Three Species of Seaweeds)

  • 양혜원;김은아;김서영;전유진
    • 한국수산과학회지
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    • 제51권1호
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    • pp.31-41
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    • 2018
  • Seaweeds are composed of a variety of bioactive substances, including polysaccharides, pigments, minerals, peptides, and polyphenols. Among these substances, the arsenic content of seaweeds has been a significant cause for concern. The present study evaluated the toxicity of arsenic from three species of seaweed using a zebrafish Danio rerio model. The arsenic-rich extracts were obtained from Ecklonia cava (ECAE), Undaria pinnatifida (UPAE) and Hizikia fusiformis (HFAE) using a solvent of 50% methanol and 1% $HNO_3$. We investigated the toxicity of the arsenic-rich extracts in zebrafish embryos through survival rate, heart rate, yolk sac edema size, cell death, reactive oxygen species (ROS) production and real-time polymerase chain reaction (PCR). The hepatotoxicity of arsenic-rich extracts was assessed in the liver of adult zebrafish through real-time PCR and histopathology. The survival rates of embryos and adult zebrafish showed no significant changes at any concentration. At 100 ppm, embryos did not exhibit significant differences in heart rate, yolk sac edema size, cell death or ROS production. In addition, apoptosis-related genes in larvae and liver tissue were unaffected by treatment with arsenic-rich extracts. These data will help clarify that developmental changes, hepatic oxidative stress, and apoptosis are not associated with toxicity from arsenic-rich seaweed extracts in a zebrafish model.

Antitumor Effects of Water Extracts of Panax notoginseng on NCI-H460 Tumor Regression Model

  • Park, Seung-Chan;Jeong, Tae-Young;Cho, Chong-Kwan;Lee, Yeon-Weol;Yoo, Hwa-Seung
    • 대한한의학회지
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    • 제31권3호
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    • pp.8-16
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    • 2010
  • Objective: This study aimed to investigate the antitumor effects of water extracts of Panax notoginseng (WEPN) in NCI-H460 human lung cancer cell xenografted nude mice. Materials and Methods: We cultured NCI-H460 cell lines and xenografted them to nude mice. The mice were divided into 3 groups; positive control group, NCI-H460+150 mg/kg WEPN-treated group, and NCI-H460+300 mg/kg WEPN-treated group. They had been raised and treated in 28 days. We checked their body weight and tumor weight and volumes twice a week and their absolute organ weight and microhistological observation at the final day. We also calculated their tumor inhibition rate (I.R.), mean survival time and percent increase in life span (% ILS). Results: Body weight of WEPN (300 mg/kg) treated mice tended to slightly greater increase than those of the positive control group, but had no significance. Tumor volume (measurement with a caliper) of WEPN-treated mice tended to be lower than that of the positive control group. Inhibition rate (I.R.) of the WEPN group decreased more than the positive control group, but had no significance. Results of tumor weights and volume (plethysmography) had no significance. Mean survival time and percent increase in life span (% ILS) in the WEPN 300 mg/kg treatment group were higher than those of any other group (p<0.05). In absolute organ weights, the WEPN (150-300 mg/kg) treatment group decreased liver weights (p<0.05). Liver tissue of mice treated with WEPN (300 mg/kg) did not show any specific lesions. Conclusion: We suggest that WEPN may have potential as a growth inhibitor of solid tumors induced by NCI-H460 without any side effects. However, this study has limitations in proving anti-tumor effects of WEPN, so further studies to overcome those limitations will be needed.

실비산의 항산화 및 지방축적 억제 효과 (Anti-oxidation and Fat Accumulation Inhibitory Effects of Silbi-san)

  • 김건우;우창훈;김영준;안희덕
    • 한방재활의학과학회지
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    • 제30권4호
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    • pp.41-53
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    • 2020
  • Objectives The purpose of this study was to investigate the effects of Silbi-san on the antioxidant and fat accumulation inhibition and to analyze the anti-obesity effect by analyzing the changes in serum lipid composition in obese mice. Methods We compared contents of phytochemicals like total polyphenols and total flavonoid and antioxidant activities such as 2,2-dipheny-1-picrylhydrazyl and 2.2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging activity. After Silbi-san in 3T3-L1 cells in vitro and mouse adipose tissue ex vivo, we quantified intracellular triglyceride accumulation and lipolysis. Moreover, the anti-obesity activity though inhibiting pancreatic lipase were analyzed. In 3T3-L1 cells, morphological changes showed that control cells had many lipid while cells treated with Silbi-san had less lipid accumulation. 30% EtOH Silbi-san treatment also suppressed the fat absorption by inhibiting the activity of pancreatic lipase and led to high lipolysis through promoting glycerol release. The experimental group was divided into four groups: Normal group fed normal feed, Control group fed 60% high fat diet (HFD) and distilled water, drug group fed 60% high fat diet and 200 mg/kg of Silbi-san water extract, drug group fed 60% HFD and 200 mg/kg of Silbi-san 30% ethanol extract. Results Serum total cholesterol content and serum low density lipoprotein-cholesterol content were significantly decreased in the Silbi-san extract group compared to the control group, serum high density lipoprotein-cholesterol content was significantly increased in Silbi-san extract group. Conclusions In this study, the antioxidant and fat accumulation inhibitory effects of Silbi-san were confirmed.

LPS로 유도된 RAW264.7 Cell과 마우스모델에 대한 잘피 에탄올 추출물의 항염증 효과 (Anti-inflammatory Effect of Zostera marina Ethanolic Extract on LPS-induced RAW264.7 Cells and Mouse Model)

  • 김민지;배난영;김꽃봉우리;박지혜;박선희;조영제;안동현
    • KSBB Journal
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    • 제30권4호
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    • pp.182-190
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    • 2015
  • The Zostera marina ethanolic extract (ZMEE) was tested in this study to investigate the anti-inflammatory activity in LPS-induced RAW 264.7 cells and mouse model. Nitric oxide production and inducible nitiric oxide synthase expression in cells treated with ZMEE was reduced significantly in a dose-dependent manner. Similarly, the secretion of pro-inflammatory cytokines such as interleukin (IL)-6, IL-$1{\beta}$, and TNF-${\alpha}$ was inhibited markedly. In addition, the expression of nuclear factor kappa B (NF-${\kappa}B$) and the phosphorylation of JNK, ERK, and p38 MAPKs was suppressed by ZMEE as well. In vivo test, ZMEE attenuated the croton oil-induced mouse ear edema and there were no mortalities in mice administered 5,000 mg/kg body weight of ZMEE during the observation periods. The results in photomicrograph of mice ear tissue showed the reduction of dermal thickness and the number of infiltrated mast cells. These results indicate that ZMEE inhibits the production of LPS-induced pro-inflammatory mediators, suggesting that ZMEE may be a potential material for anti-inflammatory therapies.

In vivo Pathogenicity Test of Oak Wilt Fungus (Raffaelea quercus-mongolicae) on Oriental Chestnut Oak (Quercus acutissima)

  • Yi, Su Hee;Lee, Jin Heung;Seo, Sang Tae;Lee, Jong Kyu
    • Journal of Forest and Environmental Science
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    • 제33권4호
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    • pp.342-347
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    • 2017
  • Since the first report of the oak wilt disease at 2004 in Korea, the disease distributed over Korean peninsula and are still giving severe damages. The management of oak wilt disease in Korea has mainly focused on the control of insect vector, Platypus koryoensis. Neverthless the effective method for evaluating the pathogenicity of the pathogen, Raffaelea quercus-mongolicae (Rqm), and for screening chemical or biological agents with strong inhibitory activity against the pathogen, is absolutely necessary, an reliable method is not available so far. This study was conducted to develop the effective method for evaluating the pathogenicity of Rqm in oak trees. The culture suspensions of Rqm were artificially injected to the saplings of Quercus acutissima by using ChemJet tree injector. Three months after treatments, the treated saplings were cut and dipped into 1% fuchsin acid solution. There were significant differences in non-conductive area (%), discoloration area (%) and vertical discoloration length between the pathogen-injected and distilled water-injected control treatments. These results indicated that the pathogen is the causal agent for the dysfunction of water conductive tissue, which will finally result in wilt symptom. Re-isolation of the pathogen and PCR detection using specific primers for the pathogen also confirmed the presence of Rqm in the sapwood chips of the pathogen-injected saplings. These observations would be greatly applied to other related researches for evaluating the pathogenicity of tree wilt pathogens and biocontrol efficacy of the selected antagonistic microorganisms, in case that the wilt symptom is not easily shown by artificial inoculation of the causal agent.

기내 배양환경이 거베라 유묘의 기외이식 후 생육에 미치는 영향 (Effects of Several Culture Conditions on in vivo Growth and Development in Gerbera hybrida)

  • 이현숙;임기병;정재동;김창길
    • 식물조직배양학회지
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    • 제28권2호
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    • pp.91-95
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    • 2001
  • 배양환경을 달리하여 생산한 배양묘 (타가영양묘, 혼합영양묘, 자가영양묘)를 기외 이식한 후에 생존율은 타가영양묘에 비해 혼합영양묘와 자가영양묘가 높았고 특히 자가영양묘는 97.6%로 가장 높았으며 정식 일수가 경과함에 따라 엽병장, 엽수, 엽면적은 자가영양묘가 타가영양묘와 혼합영양묘에 비하여 다소 좋았으나 유의성은 없었다. 정식 30일 후 유묘의 총광합성량은 자가영양묘가 154 mg$CO_2$.mgDW$^{-1}$h$^{-1}$로서 혼합영양묘 (148 mg$CO_2$.mgDW$^{-1}$h$^{-1}$)와 타가영양묘 (102 mg$CO_2$.mgDW$^{-1}$h$^{-1}$)보다 높았다. 그리고 개화시기는 자가 영양묘가 타가영양묘와 혼합영양묘에 비해 7~10일 정도가 빨랐고 엽병장, 엽수, 엽면적, 엽록소 함량 등도 양호하였다.

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The hyaluronan synthesis inhibitor 7-hydroxy-4-methylcoumarin inhibits LPS-induced inflammatory response in RAW 264.7 macrophage cells

  • Kim, Gwan Bo;Hyun, Chang-Gu
    • Journal of Applied Biological Chemistry
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    • 제64권3호
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    • pp.263-268
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    • 2021
  • 7-Hydroxy-4-methylcoumarin (7H-4MC) inhibits hyaluronan production in multiple cell lines and tissue types both in vitro and in vivo. It is a commercially available drug approved for human use, called hymecromone, in European and Asian countries to prevent biliary spasms. Nevertheless, as the pharmacological efficacy of 7H-4MC has not yet been reported in macrophages, this study investigated its anti-inflammatory effects and mechanism of action using lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. LPS-induced RAW 264.7 cells were treated with various concentrations of 7H-4MC (62.5, 125, 250, and 500 μM). The application of 7H-4MC significantly reduced nitric oxide and prostaglandin E2 production without cytotoxic effects. Additionally, 7H-4MC strongly decreased the expression of inducible nitric oxide synthase and cyclooxygenase. Furthermore, 7H-4MC reduced the production of proinflammatory cytokines, such as tumor necrosis factor-α, interleukin (IL)-1β, and IL-6. Finally, 7H-4MC exerted its potent anti-inflammatory actions via the upregulation of IκB-α production, which led to the inhibition of nuclear factor-κB (NF-κB) activity. These results, obtained in macrophage cell lines, suggest that 7H-4MC prevents inflammatory diseases via the NF-κB signaling pathway and that its use could be beneficial for human health. Ultimately, this is the first report describing the anti-inflammatory activity of 7H-4MC in a macrophage cell line.

복합생약 HO-Series의 숙취개선 임상적 유용성 평가 (Evaluation of Clinical Usefulness of Herbal Mixture HO-Series for Improving Hangover)

  • 장보윤;배진혜;김다은;김대성;조형권;김성연
    • 생약학회지
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    • 제51권4호
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    • pp.278-290
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    • 2020
  • The purpose of this study is to investigate the hangover relieving effect of HO-series. HO-S1 is an herbal mixture, which consists of extracts from Flower of Pueraria lobata Ohwi, Glycyrrhiza glabra Linné, Fruit of Lycium chinense Miller, Poria cocos Wolf, Acanthopanax sessiliflorum Seeman, Scutellaria baicalensis Georgi, Atractylodes lancea De Candlle and Zingiber officinale Roscoe. HO-S2 is a candidate that has been performed to ultra filtration based on HO-S1. HO-S3 is a mixture of amino acids and vitamins based on HO-S2. HO-01 is the final beverage base produced based on HO-S3. The antioxidant activity of HO-series was similar to that of vitamin C or trolox. The production of t-BHP induced reactive oxygen species(ROS) was significantly blocked in the presence of HO-series. In vivo study, AUC of alcohol and acetaldehyde concentrations in HO-S2 and HO-S3 treated groups significantly decreased. Hepatic alcohol dehydrogenase(ADH) and acetaldehyde dehydrogenase(ALDH) activity were significantly higher in HO-S2 and HO-S3 treated groups. And 2E1 activity and glutathione were significantly elevated, while the malondialdehyde level was not significantly in liver tissue. After alcohol exposure, the sensitivity scores of blood alcohol and acetaldehyde concentration and hangover symptoms were significantly decreased in the HO-01 intake group compared with the non-intake group. ALDH activity was significantly increased in the HO-01 intake group. HO-series have antioxidant activity and a protective effect from ROS. HO-S2, HO-S3 and HO-01 are potentially highly beneficial in relieving hangover, as it scavenges reactive free radicals and boosts the endogenous antioxidant system.