• Title/Summary/Keyword: ex vivo study

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Mitigation effects of red Platycodon grandiflorum extract on lipopolysaccharide-induced inflammation in splenocytes isolated from mice (홍도라지 추출물이 마우스에서 분리한 비장세포에서 lipopolysaccharide로 유도된 염증에 미치는 영향)

  • Park, Eun-Jung;Lee, You-Suk;Jeong, Hyun Cheol;Lee, Sung-Hyen;Lee, Hae-Jeung
    • Journal of Nutrition and Health
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    • v.52 no.3
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    • pp.243-249
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    • 2019
  • Purpose: Platycodon grandiflorum (PG) is known to have effective antimicrobial and anticancer activity. The main bioactive components of PG are saponins, and these could contribute to anti-inflammatory activity. However, little is known about the anti-inflammatory effect of PG. In this study, we aim to assess the anti-inflammatory response to Red PG Extract (RPGE) in splenocytes under ex vivo conditions. Methods: The cell viability of isolated splenocytes taken from mice was analyzed by performing a Cell Counting Kit-8 assay. The productions of nitric oxide (NO) and cytokines (specifically interleukin-6 (IL-6) and interleukin-10 (IL-10)) were measured utilizing Griess reagent and ELISA, respectively. Results: We found that co-treatment with RPGE and Lipopolysaccharide (LPS) decreased isolated splenocyte proliferation as compared with that of the LPS-stimulated control. We also observed that RPGE markedly suppressed NO synthesis and IL-6 production that was induced by LPS. There were no significant differences of IL-10 production between co-treatment with RPGE plus LPS and treatment with LPS alone. Conclusion: When taken together, our data has shown that RPGE mitigates LPS-induced inflammation in splenocytes isolated from mice. Further research is surely needed to confirm the anti-inflammation effects of RPGE in an in vivo model.

Mechanical Stretch-Induced Protection against Myocardial Ischemia-Reperfusion Injury Involves AMP-Activated Protein Kinase

  • Hao, Jia;Kim, Hun-Sik;Choi, Woong;Ha, Tae-Sun;Ahn, Hee-Yul;Kim, Chan-Hyung
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.1
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    • pp.1-9
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    • 2010
  • AMP-activated protein kinase (AMPK) protects various tissues and cells from ischemic insults and is activated by many stimuli including mechanical stretch. Therefore, this study investigated if the activation of AMPK is involved in stretch-induced cardioprotection (SIC). Intraventricular balloon and aorto-caval shunt (ACS) were used to stretch rat hearts ex vivo and in vivo, respectively. Stretch preconditioning reduced myocardial infarct induced by ischemia-reperfusion (I/R) and improved post-ischemic functional recovery. Phosphorylation of AMPK and its downstream substrate, acetyl-CoA carboxylase (ACC) were increased by mechanical stretch and ACC phosphorylation was completely blocked by the AMPK inhibitor, Compound C. AMPK activator (AICAR) mimicked SIC. Gadolinium, a blocker of stretch-activated ion channels (SACs), inhibited the stretch-induced phosphorylation of AMPK and ACC, whereas diltiazem, a specific L-type calcium channel blocker, did not affect AMPK activation. Furthermore, SIC was abrogated by Compound C and gadolinium. The in vivo stretch induced by ACS increased AMPK activation and reduced myocardial infarct. These findings indicate that stretch preconditioning can induce the cardioprotection against I/R injury, and activation of AMPK plays an important role in SIC, which might be mediated by SACs.

A Synthetic Tul4 and FopA Peptide Cocktail of Francisella tularensis Induces Humoral and Cell-Mediated Immune Responses in Mice

  • Oh, Hanseul;Kim, C-Yoon;Kim, Chang-Hwan;Hur, Gyeung-Haeng;Park, Jae-Hak
    • Journal of Microbiology and Biotechnology
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    • v.26 no.9
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    • pp.1613-1619
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    • 2016
  • Francisella tularensis is a highly virulent pathogen of humans and other mammals. Moreover, F. tularensis has been designated a category A biothreat agent, and there is growing interest in the development of a protective vaccine. In the present study, we determine the in vitro and in vivo immune responses of a subunit vaccine composed of recombinant peptides Tul4 and FopA from epitopes of the F. tularensis outer membrane proteins. The recombinant peptides with adjuvant CpG induced robust immunophenotypic change of dendritic cell (DC) maturation and secretion of inflammatory cytokines (IL-6, IL-12). In addition, the matured DCs enabled ex vivo proliferation of naive splenocytes in a mixed lymphocyte reaction. Lastly, we determined the in vivo immune response by assessment of antibody production in C57BL/6 mice. Total IgG levels were produced after immunization and peaked in 6 weeks, and moreover, Tul4-specific IgG was confirmed in the mice receiving peptides with or without CpG. Based on these results, we concluded that the recombinant peptides Tul4 and FopA have immunogenicity and could be a safe subunit vaccine candidate approach against F. tularensis.

Moisturization and Transdermal Penetration Characteristics of PEGimpregnated Aloe vera Gel from DIS Processing (DIS에 의한 Polyethylene Glycol 함침 알로에 베라 겔의 보습 및 경피흡수 특성)

  • Kwon, Hye Mi;Hur, Won;Lee, Shin Young
    • KSBB Journal
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    • v.28 no.5
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    • pp.319-326
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    • 2013
  • This study was carried out to investigate the in vitro and in vivo moisturizing properties and percutaneous absorption of PEG-impregnated Aloe vera gel. The PEG-i-Aloe gel was obtained from dewatering and impregnation by soaking (DIS) of Aloe vera leaf slice. The moisturizing property of the obtained sample was evaluated by moisture determination using gravimetric method in desiccator under different RH% and by water sorption-desorption test on human skin. The transdermal penetration characteristics of PEG-i-Aloe gel was investigated by Franz diffusion cell in vitro transdermal absorption method. PEG-i-Aloe gel had high moisture retention ability and could significantly lead the enhancing skin hydration status as well as reducing the skin water loss due to the film formation as a skin barrier. The skin penetration rate of PEGi- Aloe gel at steady state was 9.76 ${\mu}g/(h{\cdot}cm^2)$ and the quantity of the transdermal absorption was 144 ${\mu}g/cm^2$ in 9 hr. The penetration mechanism was well fitted with Higuchi model ($R^2$ = 0.974-0.994). The results show that PEG-i-Aloe gel has the significant moisturizing effect and strong penetration of the animal skin. It could be used as the moisturizing additive in cosmetic skin products.

Water soluble tomato concentrate regulates platelet function via the mitogen-activated protein kinase pathway

  • Jeong, Dahye;Irfan, Muhammad;Saba, Evelyn;Kim, Sung-Dae;Kim, Seung-Hyung;Rhee, Man Hee
    • Korean Journal of Veterinary Research
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    • v.56 no.2
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    • pp.67-74
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    • 2016
  • Tomato extract has been shown to exert antiplatelet activity in vitro and to change platelet function ex vivo, but with limitations. In this study, antiplatelet activity of water soluble tomato concentrate (Fruitflow I) and dry water soluble tomato concentrate (Fruitflow II) was investigated using rat platelets. Aggregation was induced by collagen and adenosine diphosphate and granule-secretion, $[Ca^{2+}]_i$, thromboxane B2, cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) levels were examined. The activation of integrin ${\alpha}_{IIb}{\beta}_3$ and phosphorylation of signaling molecules, including mitogen-activated protein kinase (MAPK) and PI3K/Akt, were investigated by flow cytometry and immunoblotting, respectively. Prothrombin time (PT) and activated partial thromboplastin time (aPTT) were examined. Moreover, in vivo thrombus weight was tested by an arteriovenous shunt model. Fruitflow I and Fruitflow II significantly inhibited agonist induced platelet aggregation, adenosine triphosphate and serotonin release, $[Ca^{2+}]_i$, and thromboxane B2 concentration, while having no effect on cAMP and cGMP levels. Integrin ${\alpha}_{IIb}{\beta}_3$ activation was also significantly decreased. Moreover, both concentrates reduced phosphorylation of MAPK pathway factors such as ERK, JNK, P38, and PI3K/Akt. In vivo thrombus formation was also inhibited. Taken together, these concentrates have the potential for ethnomedicinal applications to prevent cardiovascular ailments and can be used as functional foods.

Effects of Indomethacin on the Production of Cytokines in Mice Exposed to Excessive Zinc (과량의 아연에 노출된 생쥐의 사이토카인 생산에 미치는 인도메타신의 영향)

  • 채병숙;신태용
    • YAKHAK HOEJI
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    • v.46 no.4
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    • pp.258-264
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    • 2002
  • Zinc plays an important role in immunobiological responses, while excessive zinc attenuates immune functions in a dose-dependent manner. Zinc excess has been reported to increase levels of plasma prostaglandin E$_2$ (PGE$_2$), which is known to inhibit production of Th (helper T) 1-associated cytokines and to induce inflammatory responses. Thus, this study was investigated the effects of indomethacin, a potent inhibitor of PGE$_2$ synthesis, on the proinflammatory cytokine and lymphokine production in ICR mice exposed to excessive zinc. Indomethacin at doses of 5 mg/kg was administered i.p. 30 minutes before zinc chloride (Zn) 30 mg/kg orally daily for 10 days. Excessive Zn remarkedly increased tumor necrosis factor (TNF)-$\alpha$ and interleukin (IL)-1$\beta$ levels in both serum and splenic supernatants compared with those in controls, while indomethacin significantly reduced the excessive Zn-induced levels of IL-1$\beta$. In serum, excessive Zn significantly decreased the levels of IL-2 and interferon (IFN)-${\gamma}$ compared with those in controls, whereas indomethacin significantly enhanced the excessive Zn-decreased levels of IFN-${\gamma}$ but did not affect the Zn-decreased levels of serum IL-2. In splenic supernatants, All of excessive Zn, indomethacin, and combination of Zn and indomethacin significantly enhanced IL-2 levels compared with those in controls, but indomethacin didn't affect the Zn-induced production of IL-2. These data, therefore, suggest that indomethacin significantly attenuated the in vivo and ex vivo IL-1$\beta$ production increased by excessive zinc and remarkedly enhanced the in vivo excessive zinc-suppressed production of IFN-${\gamma}$ but not IL-2.

Oligonol promotes anti-aging pathways via modulation of SIRT1-AMPK-Autophagy Pathway

  • Park, Seul-Ki;Seong, Rak-Kyun;Kim, Ji-Ae;Son, Seok-Jun;Kim, Younghoon;Yokozawa, Takako;Shin, Ok Sarah
    • Nutrition Research and Practice
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    • v.10 no.1
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    • pp.3-10
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    • 2016
  • BACKGROUND/OBJECTIVES: Oligonol, mainly found in lychee fruit, is an antioxidant polyphenolic compound which has been shown to have anti-inflammatory and anti-cancer properties. The detailed mechanisms by which oligonol may act as an anti-aging molecule have not been determined. MATERIALS/METHODS: In this study, we evaluated the ability of oligonol to modulate sirtuin (SIRT) expression in human lung epithelial (A549) cells. Oligonol was added to A549 cells and reactive oxygen species production, mitochondrial superoxide formation, and p21 protein levels were measured. Signaling pathways activated upon oligonol treatment were also determined by western blotting. Furthermore, the anti-aging effect of oligonol was evaluated ex vivo in mouse splenocytes and in vivo in Caenorhabditis elegans. RESULTS: Oligonol specifically induced the expression of SIRT1, whose activity is linked to gene expression, metabolic control, and healthy aging. In response to influenza virus infection of A549 cells, oligonol treatment significantly up-regulated SIRT1 expression and down-regulated viral hemagglutinin expression. Oligonol treatment also resulted in the activation of autophagy pathways and the phosphorylation of AMP-activated protein kinase (AMPK). Furthermore, oligonol-treated spleen lymphocytes from old mice showed increased cell proliferation, and mRNA levels of SIRT1 in the lungs of old mice were significantly lower than those in the lungs of young mice. Additionally, in vivo lethality assay revealed that oligonol extended the lifespan of C. elegans infected with lethal Vibrio cholerae. CONCLUSIONS: These data demonstrated that oligonol may act as an anti-aging molecule by modulating SIRT1/autophagy/AMPK pathways.

Comparison between the antioxidant properties of environment-friendly agro-produce and processed food

  • Kim, Soo-Mi;Kim, Joo-Hee;Son, Myoung-Jin;Kim, Se-Young;Rico, Catherine;Kang, Mi-Young
    • Proceedings of the Korean Society of Organic Agriculture Conference
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    • 2009.12a
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    • pp.316-316
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    • 2009
  • This study was conducted to investigate the effects of environment-friendly materials on garlic cultivation, and the methods for processing garlic at proper ripening stage to increase its antioxidant properties and market value. The functional properties of garlic and processed black garlic were analyzed in vitro and ex vivo. The ethanol extracts of garlic and black garlic were used in the antioxidant, linoleic acid autooxidation, lipid oxidation and RAW264.7 indicator cell using NO induced activity analyses. Also, toxicity tests by RBL-2H3 and RAW264.7 indicator cell using MTT assay were compared. Obesity inhibition test of garlic and black garlic were also studied.

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Immortalization of Primary Keratinocytes and Its Application to Skin Research

  • Choi, Moonju;Lee, Choongho
    • Biomolecules & Therapeutics
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    • v.23 no.5
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    • pp.391-399
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    • 2015
  • As a major component of the epidermal tissue, a primary keratinocyte has served as an essential tool not only for the study of pathogenesis of skin-related diseases but also for the assessment of potential toxicities of various chemicals used in cosmetics. However, its short lifespan in ex vivo setting has been a great hurdle for many practical applications. Therefore, a number of immortalization attempts have been made with success to overcome this limitation. In order to understand the immortalization process of a primary keratinocyte, several key biological phenomena governing its lifespan will be reviewed first. Then, various immortalization methods for the establishment of stable keratinocyte cell lines will be explained. Finally, its application to a three-dimensional skin culture system will be described.

Belamcandae Rhizoma Extract Improves Memory Impairment in Mice (생쥐에서 사간 추출물의 기억력 감퇴에 대한 개선 효과)

  • Jung, Ji Wook
    • Korean Journal of Pharmacognosy
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    • v.52 no.1
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    • pp.55-60
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    • 2021
  • Belamcandae Rhizoma (BR), the rhizome of Belamcanda chinensis (L.) DC., possesses various biological properties such as anti-inflammatory activity, antioxidant activity and antimutagenic activity. However, there have been no studies on the anti-amnesic effect of BR. In this study, we assessed the improvement effect of BR extract on scopolamine-induced amnesia in mice. ICR mice were administrated with BR (50, 100 or 200 mg/kg, p.o.) and were subsequently injected of scopolamine (1 mg/kg, i.p.) 30 min before behavioral tasks (Y-maze, passive avoidance and Morris water maze tasks). To further assess the possible mechanisms of BR, the ex vivo acetylcholinesterase (AChE) activity was also evaluated. BR could ameliorate scopolamine-induced memory impairment and could regulate the cholinergic function by inhibiting the AChE activity. These data demonstrated that BR exert candidate extract against amnesia by restoring the cholinergic activity.