• Molecules and Cells
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• 제43권3호
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• pp.236-250
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• 2020

Currently, many available anti-cancer therapies are targeting apoptosis. However, many cancer cells have acquired resistance to apoptosis. To overcome this problem, simultaneous induction of other types of programmed cell death in addition to apoptosis of cancer cells might be an attractive strategy. For this purpose, we initially investigated the inhibitory role of TRIP-Br1/XIAP in necroptosis, a regulated form of necrosis, under nutrient/serum starvation. Our data showed that necroptosis was significantly induced in all tested 9 different types of cancer cell lines in response to prolonged serum starvation. Among them, necroptosis was induced at a relatively lower level in MCF-7 breast cancer line that was highly resistant to apoptosis than that in other cancer cell lines. Interestingly, TRIP-Br1 oncogenic protein level was found to be very high in this cell line. Up-regulated TRIP-Br1 suppressed necroptosis by repressing reactive oxygen species generation. Such suppression of necroptosis was greatly enhanced by XIAP, a potent inhibitor of apoptosis. Our data also showed that TRIP-Br1 increased XIAP phosphorylation at serine87, an active form of XIAP. Our mitochondrial fractionation data revealed that TRIP-Br1 protein level was greatly increased in the mitochondria upon serum starvation. It suppressed the export of CypD, a vital regulator in mitochondria-mediated necroptosis, from mitochondria to cytosol. TRIP-Br1 also suppressed shikonin-mediated necroptosis, but not TNF-α-mediated necroptosis, implying possible presence of another signaling pathway in necroptosis. Taken together, our results suggest that TRIP-Br1/XIAP can function as onco-proteins by suppressing necroptosis of cancer cells under nutrient/serum starvation.

• Journal of Microbiology and Biotechnology
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• 제26권6호
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• pp.1035-1045
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• 2016

To evaluate the effects of lactic acid bacteria (LAB) on Peyer's patch cells, mice were treated with a high dose of kanamycin to disturb the gut microbial environment. The overarching goal was to explore the potential of LAB for use as a dietary probiotic that buffers the negative consequences of antibiotic treatment. In vitro, LAB stimulated the production of immunoglobulin A (IgA) from isolated Peyer's patch cells. Inflammation-related genes (TNF-α, IL-1β, and IL-8) were up-regulated in Caco-2 cells stimulated with lipopolysaccharide (LPS), while tight-junction-related genes (ZO-1 and occludin) were down-regulated; the effects of LPS on inflammatory gene and tight-junction gene expression were reversed by treatment with LAB. Mice treated with a high dose of kanamycin showed increased serum IgE levels and decreases in serum IgA and fecal IgA levels; the number of Peyer's patch cells decreased with kanamycin treatment. However, subsequent LAB treatment was effective in reducing the serum IgE level and recovering the serum IgA and fecal IgA levels, as well as the number of Peyer's patch cells. In addition, ZO-1 and occludin mRNA levels were up-regulated in the ileum tissues of mice receiving LAB treatment. Lactic acid bacteria can enhance the intestinal immune system by improving the integrity of the intestinal barrier and increasing the production of IgA in Peyer's patches. Lactic acid bacteria should be considered a potential probiotic candidate for improving intestinal immunity, particularly in mitigating the negative consequences of antibiotic use.

• 대한한방내과학회지
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• 제41권3호
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• pp.339-349
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• 2020

Objective: This study aimed to evaluate anti-inflammatory and antitussive expectoration effects of Friltillariae Thunbergii Bulbus (FTB) in a mouse model. Materials and Methods: To evaluate the anti-inflammatory effects of the FTB, we conducted in vitro experiments using RAW264.7 cells. An MTT assay and enzyme-linked immunosorbent assay (ELISA) were carried out to examine the anti-inflammatory effects of FTB. The expectorant effect on phenol red secretion, the antitussive effect on cough induced by ammonia solution, and leukocyte increased inhibition effects in acute airway inflammation in the animal model were confirmed. Results: FTB did not show cytotoxicity in the experimental group at 10, 30, 50, 100, 300, or 500 ㎍/ml and significantly inhibited the increase of NO, TNF-α and IL-6 in the experimental groups at 30, 50, 100, 300, and 500 ㎍/ml concentrations. In sputum, cough, and acute airway inflammation animal models, FTB significantly increased phenol red secretion in the 400 mg/kg administration group. FTB significantly reduced the number of coughs and significantly increased cough delay time in both 200 and 400 mg/kg dose groups. FTB decreased the white blood cell count in BALF (bronchoalveolar lavage fluid) in the 400 mg/kg administration group. Conclusion: Our study revealed that FTB elicits antitussive and expectorant effects by inhibiting inflammatory cytokines, increasing sputum secretion, suppressing cough, and reducing inflammatory cells. We concluded that FTB is a highly promising agent for respiratory tract infection with therapeutic opportunities.

• Biomolecules & Therapeutics
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• 제28권5호
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• pp.456-464
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• 2020

Mast cells (MCs) are systemically distributed and secrete several allergic mediators such as histamine and leukotrienes to cause type I hypersensitivity. Dasatinib is a type of anti-cancer agent and it has also been reported to inhibit human basophils. However, dasatinib has not been reported for its inhibitory effects on MCs or type I hypersensitivity in mice. In this study, we examined the inhibitory effect of dasatinib on MCs and MC-mediated allergic response in vitro and in vivo. In vitro, dasatinib inhibited the degranulation of MCs by antigen stimulation in a dose-dependent manner (IC₅₀, ~34 nM for RBL-2H3 cells; ~52 nM for BMMCs) without any cytotoxicity. It also suppressed the secretion of inflammatory cytokines IL-4 and TNF-α by antigen stimulation. Furthermore, dasatinib inhibited MC-mediated passive cutaneous anaphylaxis (PCA) in mice (ED₅₀, ~29 mg/kg). Notably, dasatinib significantly suppressed the degranulation of MCs in the ear tissue. As the mechanism of its effect, dasatinib inhibited the activation of Syk and Syk-mediated downstream signaling proteins, LAT, PLCγ1, and three typical MAP kinases (Erk1/2, JNK, and p38), which are essential for the activation of MCs. Interestingly, in vitro tyrosine kinase assay, dasatinib directly inhibited the activities of Lyn and Fyn, the upstream tyrosine kinases of Syk in MCs. Taken together, dasatinib suppresses MCs and PCA in vitro and in vivo through the inhibition of Lyn and Fyn Src-family kinases. Therefore, we suggest the possibility of repositioning the anti-cancer drug dasatinib as a treatment for various MC-mediated type I hypersensitive diseases.

• 한국식품과학회지
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• 제52권2호
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• pp.138-143
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• 2020

본 연구에서는 다양한 만성 대사성 질환에 기초적 유발 기전인 산화적 스트레스 및 염증에 대한 폴리페놀 함유 블랙 초크베리의 보호 효과를 살펴보았다. 블랙 초크베리는 DPPH, ·OH, 및 NO 등 다양한 라디컬 소거능을 나타냈다. 내독소인 LPS에 의해 활성화된 대식세포 내 ROS 증가는 폴리페놀 함유 블랙 초크베리에 의해 감소하였다. 이는 블랙 초크베리가 전사인자인 NRF2의 핵 내 이동 및 이로 인해 조절되는 SOD에 의한 ROS 소거능 및 NOX2에 의한 ROS 생성 저하에 의한 것으로 나타났다. 블랙 초크베리의 염증 보호 효과는 활성화된 대식세포에서 증가하는 NF-κB의 핵 내 이동에 따른 COX-2, iNOS와 같은 염증 매개인자 관련 효소와 TNF-α, IL-6, IL-1β와 같은 염증성 사이토카인의 저하에 의한 것으로 나타났다. 결론적으로 블랙 초크베리의 산화적 스트레스 및 염증 보호 효과는 항산화 효소 관련 전사 인자인 NRF2 조절 기전 및 염증 관련 전사 인자인 NF-κB와 관련 유전자 발현의 조절을 통하여 나타났다. 추후 블랙 초크베리 내 기능성 물질에 대한 심도 있는 기전 연구를 통해 천연물 유래 기능성 소재로서의 역할을 할 것으로 판단된다.

• 대한한방부인과학회지
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• 제29권1호
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• pp.1-13
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• 2016

Objectives The purpose of this study is to find out the effect of various concentrations of Hwangnyeonhaedok-tang (HTT) pharmacopuncture on ST25 (天樞) in rats with dextran sulfate sodium (DSS)-induced colitis.Methods Colitis was experimentally induced by feeding rats with water mixed with 5% (w/v) DSS for 20 days. The rats were divided into 5 groups: the normal group (Nor, n=5), the control group - colitis induced rats with no treatment (Con, n=5), the acupuncture group - colitis induced rats with acupuncture applied on ST25 (Acu, n=5), the pharmacopuncture group 1 - colitis induced rats with 0.729 mg/250 g/40 μl of pharmacopuncture applied on ST25 (PA-1, n=5), the pharmacopuncture group 2 - colitis induced rats with 3.645 mg/250 g/40 μl of pharmacopuncture applied on ST25 (PA-2, n=5). The changes in weight, excrement concentration and hemafecia were observed 5 times every 2 days. The colon lengths were measured from appendix to the end of colon after the experiment. Hematological and serological exams were conducted the day after the last treatment by cardiac puncturing anesthetized rats.Results ST25 is the abdominal front point (募穴) of large intestine meridian and is known to have effect in colitis. Various concentrations of HTT pharmacopuncture (HTTP) applied on ST25, in rats with DSS-induced colitis inhibited decrease in colon lengths and body weight in both PA-1 and PA-2 groups. Hematological and serological exam results also showed that HTTP has significant effect on colitis in both PA-1 and PA-2 groups.Conclusions Colon lengths were significantly increased in the acupuncture group, PA-1 group and PA-2 group, compared to the control group. The body weight was significantly increased (p<0.05) in PA-2 group after the first treatment, compared to the control group. TNF-α, IL-6, AST were significantly decreased in PA-1 and PA-2 groups, compared to the control group.

• 한국식품영양학회지
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• 제34권5호
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• pp.487-497
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• 2021

To investigate the industrial availability of liquid fermentation (PL-ferment) by Phellinus linteus mycelium as a postbiotics for the inhibition of inflammation, PL-ferment was fractionated into culture supernatant (CS), hot-water extract (HW) from PL-ferment, EtOH-precipitate (CP) fractionated from HW, and the dialysate (DCP) of CP. Compared to the other fractions, DCP which is expected to contain exopolysaccharide (EPS) as the major component, significantly decreased the production of NO, IL-6, and MCP-1 in LPS-induced RAW 264.7 cells, and IL-6 and IL-8 in TNF-α and IFN-γ-induced HaCaT cells. The general component analysis results showed that no significant difference in components was observed between the fractions, whereas sugar composition analysis revealed that DCP had decreased glucose and increased mannose contents compared to the other fractions. This suggests that mannose played an important role in the anti-inflammatory activity of the active fraction, DCP. Molecular weight distribution analysis revealed that DCP was mainly composed of low-molecular-weight material-removed high-molecular-weight polysaccharides of 18-638 kDa, suggesting that EPS originated from P. linteus EPS. In conclusion, our results suggest that the DCP of P. linteus mycelium fermentation using the anti-inflammatory activity could be used industrially as postbiotic material.

• 대한한방내과학회지
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• 제42권1호
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• pp.25-39
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• 2021

Objective: The purpose of this study was to investigate the effect of Lonicera Japonica Thunberg (LJT) on the inflammatory factor expression associated with atherosclerosis in human umbilical vein endothelial cells (HUVECs). Methods: After treatment with LJT in HUVEC which is treated with TNF-α, we measured the expression levels of biomarkers (MCP-1, ICAM-1, VCAM-1, KLF2, and eNOS), mRNA (CCL2, ICAM1, VCAM1, KLF2, and NOS3), and the proteins (MCP-1, ICAM-1, VCAM-1, KLF2, eNOS, ERK, JNK, and p38). Results: 1. Compared to the control, LJT significantly reduced MCP-1 and VCAM-1 levels at concentrations of 100, 200, and 400 ㎍/ml and ICAM-1 expression at 200 and 400 ㎍/ml compared to the control. It increased KLF2 levels at all three concentrations, but not significantly, while eNOS expression was significantly increased at 400 ㎍/ml. 2. LJT was seen to significantly reduce the expression of CCL2, ICAM1, and VCAM1 mRNA at concentrations of 100, 200, and 400 ㎍/ml compared to the control. In contrast, significantly increased KLF2 and NOS3 mRNA levels were observed at 400 ㎍/ml and at 200 and 400 ㎍/ml, respectively. 3. Compared to the control, LJT significantly reduced the protein expression of MCP-1 and VCAM-1 at 200 and 400 ㎍/ml and of ICAM-1 at 400 ㎍/ml. In addition, it increased both KLF2 and eNOS protein levels at 200 and 400 ㎍/ml. Although LJT did not have an effect on ERK expression in comparison with the control, it significantly reduced JNK levels at 200 and 400 ㎍/ml and p38 levels at 400 ㎍/ml. Conclusions: These results suggest that LJT has an effect on the inhibition of inflammatory factor expression associated with atherosclerosis in HUVECs which could contribute to the prevention of cardiovascular and cerebrovascular diseases.

• 한국유기농업학회지
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• 제28권4호
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• pp.591-604
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• 2020

지금까지 유자에서 추출한 Limonene은 항균활성 및 항산화 활성이 높은 것으로 알려져 왔으나, 면역능력 조절능력에 대한 연구가 부재하여 본 연구에서는 유자에서 Limonene 추출의 최적조건 탐색 및 대식세포와 마우스의 혈청 내 cytokine 분비능을 조사하였다. 추출의 최적조건 탐색 실험에서는 증류수를 이용한 연속증류추출법의 경우가 refluxing이라는 과정을 더해 줬을 때 다른 추출법보다 시간이 다소 오래 소요되는 단점은 있으나 용매를 이용하지 않고 증류수만 이용하여도 추출 후 Limonene의 회수율을 높일 수 있기에 편리하고 경제성이 뛰어난 추출법이라 판단되었다. 마우스의 대식세포를 이용한 in vitro 실험에서는 Limonene 처리에 의한 대식세포증식활성이 증가되는 경향을 보였으며 세포증식활성 관련 유전자 발현도 Limonene 추출물 처리에 의해 증가하였다. 마우스를 이용한 in vivo 실험에서는 대조군에 비해 사료섭취량이 다소 감소하는 경향을 나타내었으며 이 결과가 체중증가량에 반영된 것으로 나타났다. 이것은 Limonene 추출물을 21일간 직접 경구 투여한 것이 위내 자극을 유도하였을 수도 있다고 판단되었다. 한편, 마우스에 면역자극을 유도하지 않은 조건하에서 혈중 IL-1β이 Limonene 급여 농도 의존적으로 증가한 것으로 나타나 In vitro 결과를 잘 반영해 주었다. 이러한 연구 결과를 바탕으로 Limonene을 다른 영양성분과의 결합 등을 유도한 by-pass Limonene을 제조하여 면역기능 조절활성을 유도할 가능성 등의 연구가 더욱 필요하다고 사료된다. 그러나 본 연구를 통해 유자추출물인 Limonene의 면역기능 조절활성을 갖는 것이 확인됨으로서 유자추출물의 친환경 사료첨가제 소재로서 개발 가능성이 제시되었다.

• Biomolecules & Therapeutics
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• 제29권6호
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• pp.685-696
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• 2021

In this study, we investigated the inhibitory effect of 5-aminolevulinic acid (ALA), a heme precursor, on inflammatory and oxidative stress activated by lipopolysaccharide (LPS) in RAW 264.7 macrophages by estimating nitric oxide (NO), prostaglandin E2 (PGE2), cytokines, and reactive oxygen species (ROS). We also evaluated the molecular mechanisms through analysis of the expression of their regulatory genes, and further evaluated the anti-inflammatory and antioxidant efficacy of ALA against LPS in the zebrafish model. Our results indicated that ALA treatment significantly attenuated the LPS-induced release of pro-inflammatory mediators including NO and PGE2, which was associated with decreased inducible NO synthase and cyclooxygenase-2 expression. ALA also inhibited the LPS-induced expression of pro-inflammatory cytokines, such as tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6, reducing their extracellular secretion. Additionally, ALA abolished ROS generation, improved the mitochondrial mass, and enhanced the expression of heme oxygenase-1 (HO-1) and the activation of nuclear translocation of nuclear factor-E2-related factor 2 (Nrf2) in LPS-stimulated RAW 264.7 macrophages. However, zinc protoporphyrin, a specific inhibitor of HO-1, reversed the ALA-mediated inhibition of pro-inflammatory cytokines production and activation of mitochondrial function in LPS-treated RAW 264.7 macrophages. Furthermore, ALA significantly abolished the expression of LPS-induced pro-inflammatory mediators and cytokines, and showed strong protective effects against NO and ROS production in zebrafish larvae. In conclusion, our findings suggest that ALA exerts LPS-induced anti-inflammatory and antioxidant effects by upregulating the Nrf2/HO-1 signaling pathway, and that ALA can be a potential functional agent to prevent inflammatory and oxidative damage.