• Nutrition Research and Practice
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• 제7권4호
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• pp.249-255
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• 2013

In this study, the protective effects of EGCG on L-3,4-dihydroxyphenylalanine (L-DOPA)-induced oxidative cell death in catecholaminergic PC12 cells, the in vitro model of Parkinson's disease, were investigated. Treatment with L-DOPA at concentrations higher than $150{\mu}M$ caused cytotoxicity in PC12 cells, as determined using the 3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry detection. The apoptotic ratio was similar in cells treated with $100{\mu}M$ EGCG plus $150{\mu}M$ L-DOPA (5.02%) and the control (0.96%) (P > 0.05), and was lower than that of cells treated with L-DOPA only (32.24%, P < 0.05). The generation level of ROS (% of control) in cells treated with EGCG plus L-DOPA was lower than that in cells treated with L-DOPA only (123.90% vs 272.32%, P < 0.05). The optical density in production of TBARS in cells treated with L-DOPA only was higher than that in the control ($0.27{\pm}0.05$ vs $0.08{\pm}0.04$, P < 0.05), and in cells treated with EGCG only ($0.14{\pm}0.02$, P < 0.05), and EGCG plus L-DOPA ($0.13{\pm}0.02$, P < 0.05). The intracellular level of GSH in cells treated with EGCG plus L-DOPA was higher than that in cells treated with L-DOPA only ($233.25{\pm}16.44$ vs $119.23{\pm}10.25$, P < 0.05). These results suggest that EGCG protects against L-DOPA-induced oxidative apoptosis in PC12 cells, and might be a potent neuroprotective agent.

• The Korean Journal of Physiology and Pharmacology
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• 제26권1호
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• pp.25-36
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• 2022

To identify the effect and mechanism of carbon monoxide (CO) on delayed rectifier K⁺ currents (I_K) of human cardiac fibroblasts (HCFs), we used the wholecell mode patch-clamp technique. Application of CO delivered by carbon monoxidereleasing molecule-3 (CORM3) increased the amplitude of outward K⁺ currents, and diphenyl phosphine oxide-1 (a specific I_K blocker) inhibited the currents. CORM3-induced augmentation was blocked by pretreatment with nitric oxide synthase blockers (L-NG-monomethyl arginine citrate and L-NG-nitro arginine methyl ester). Pretreatment with KT5823 (a protein kinas G blocker), 1H-[1,-2,-4] oxadiazolo-[4,-3-a] quinoxalin-1-on (ODQ, a soluble guanylate cyclase blocker), KT5720 (a protein kinase A blocker), and SQ22536 (an adenylate cyclase blocker) blocked the CORM3 stimulating effect on I_K. In addition, pretreatment with SB239063 (a p38 mitogen-activated protein kinase [MAPK] blocker) and PD98059 (a p44/42 MAPK blocker) also blocked the CORM3's effect on the currents. When testing the involvement of S-nitrosylation, pretreatment of N-ethylmaleimide (a thiol-alkylating reagent) blocked CO-induced I_K activation and DL-dithiothreitol (a reducing agent) reversed this effect. Pretreatment with 5,10,15,20-tetrakis(1-methylpyridinium-4-yl)-21H,23H porphyrin manganese (III) pentachloride and manganese (III) tetrakis (4-benzoic acid) porphyrin chloride (superoxide dismutase mimetics), diphenyleneiodonium chloride (an NADPH oxidase blocker), or allopurinol (a xanthine oxidase blocker) also inhibited CO-induced I_K activation. These results suggest that CO enhances I_K in HCFs through the nitric oxide, phosphorylation by protein kinase G, protein kinase A, and MAPK, S-nitrosylation and reduction/oxidation (redox) signaling pathways.

• Journal of Nutrition and Health
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• 제56권1호
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• pp.12-23
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• 2023

Purpose: This study investigates the alterations in A549 human non-small-cell lung cancer (NSCLC) cells exposed to Citrus junos extract (CJE). We further examine the antiproliferative and apoptotic effects of CJE on NSCLC cells. Methods: Inhibition of proliferation was examined by applying the 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) colorimetric assay on CJE-treated A549 NSCLC cells. The lactate dehydrogenase (LDH) assay was performed to measure the degree of toxicity of CJE on NSCLC cells. The effect on migratory proliferation was confirmed using the scratch wound healing assay. The antiproliferative effect of the CJE on human lung cancer cells was verified through morphological observation, fluorescence microscopy, and caspase-3 colorimetry. Results: Exposure of NSCLC cells to CJE resulted in a dose- and time-dependent decrease in cell activity and increased toxicity to the cells. In addition, microscopic observation revealed a reduced ability of the cancer cells to migrate and proliferate after exposure to the CJE, with simultaneous morphological apoptotic changes. Fluorescence staining and microscopic examination revealed that this death was a process of self-programmed cell death of NSCLC cells. Compared to unexposed NSCLC cells, the expression of caspase-3 was significantly increased in cells exposed to CJE. Conclusion: Exposure of A549 human NSCLC cells to CJE inhibits the proliferation, increases the cytotoxicity, and decreases the ability of cells to migrate and grow. Moreover, the expression of caspase-3 increases after CJE treatment, suggesting that the apoptosis of NSCLC cells is induced by a chain reaction initiated by caspase-3. These results indicate that Citrus junos is a potential therapeutic agent for human non-small-cell lung cancer.

• Restorative Dentistry and Endodontics
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• 제48권1호
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• pp.2.1-2.12
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• 2023

Objectives: In this study, natural substances were introduced as primary dental pulp caps for use in pulp therapy, and the antimicrobial and cytotoxic properties of these substances were investigated. Materials and Methods: In this in vitro study, the antimicrobial properties of calcium-enriched mixture (CEM) cement, propolis, and propolis individually combined with the extracts of several medicinal plants were investigated against Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. Then, the cytotoxicity of each substance or mixture against pulp stem cells extracted from 30 primary healthy teeth was evaluated at 4 concentrations. Data were gathered via observation, and optical density values were obtained using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) test and recorded. SPSS software version 23 was used to analyze the data. Data were evaluated using 2-way analysis of variance and the Tukey test. Results: Regarding antimicrobial properties, thyme alone and thyme + propolis had the lowest minimum inhibitory concentrations (MICs) against the growth of S. aureus, E. coli, and P. aeruginosa bacteria. For E. faecalis, thyme + propolis had the lowest MIC, followed by thyme alone. At 24 and 72 hours, thyme + propolis, CEM cement, and propolis had the greatest bioviability in the primary dental pulp stem cells, and lavender + propolis had the lowest bioviability. Conclusions: Of the studied materials, thyme + propolis showed the best results in the measures of practical performance as a dental pulp cap.

• Journal of Veterinary Science
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• 제23권1호
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• pp.4.1-4.15
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• 2022

Background: Flavonoids are natural polyphenols found widely in citrus fruit and peel that possess anti-adipogenic effects. On the other hand, the detailed mechanisms for the antiadipogenic effects of flavonoids are unclear. Objectives: The present study observed the anti-adipogenic effects of five major citrus flavonoids, including hesperidin (HES), narirutin (NAR), nobiletin (NOB), sinensetin (SIN), and tangeretin (TAN), on AMP-activated protein kinase (AMPK) activation in palmitate (PA)-treated HepG2 cells. Methods: The intracellular lipid accumulation and triglyceride (TG) contents were quantified by Oil-red O staining and TG assay, respectively. The glucose uptake was assessed using 2-[N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose (2-NBDG) assay. The levels of AMPK, acetyl-CoA carboxylase (ACC), and glycogen synthase kinase 3 beta (GSK3β) phosphorylation, and levels of sterol regulatory element-binding protein 2 (SREBP-2) and 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) expression were analyzed by Western blot analysis. The potential interaction between the flavonoids and the γ-subunit of AMPK was investigated by molecular docking analysis. Results: The flavonoid treatment reduced both intracellular lipid accumulation and TG content in PA-treated HepG2 cells significantly. In addition, the flavonoids showed increased 2-NBDG uptake in an insulin-independent manner in PA-treated HepG2 cells. The flavonoids increased the AMPK, ACC, and GSK3β phosphorylation levels and decreased the SREBP-2 and HMGCR expression levels in PA-treated HepG2 cells. Molecular docking analysis showed that the flavonoids bind to the CBS domains in the regulatory γ-subunit of AMPK with high binding affinities and could serve as potential AMPK activators. Conclusion: The overall results suggest that the anti-adipogenic effect of flavonoids on PA-treated HepG2 cells results from the activation of AMPK by flavonoids.

• Animal Bioscience
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• 제36권12호
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• pp.1785-1795
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• 2023

Objective: This study was to estimate heritabilities, additive genetic correlations, and phenotypic correlations between number of piglets born alive (NBA), litter birth weight (LTBW), number of piglets weaned (NPW) and litter weaning weight (LTWW) in different parities of Landrace (L), Yorkshire (Y), Landrace×Yorkshire (LY), and Yorkshire×Landrace (YL) sows in a commercial swine operation in Northern Thailand. Methods: Two models were utilized, a single trait repeatability model (RM) and a multiple trait animal model (MTM). The RM assumed reproductive records from different parities to be repeated values of the same trait, whereas the MTM assumed these records to be different traits. The two models accounted for the fixed effects of farrowing year-season, genetic group of the sow, heterosis, and age at first farrowing, and the random effects of sow, boar, and residual. Results: Heritability estimates from RM were 0.02±0.01 for NBA, 0.10±0.01 for LTBW, 0.04±0.01 for NPW, and 0.11±0.01 for LTWW. Heritability estimates from MTM fluctuated across parities, ranging from 0.04±0.01 in parity 2 to 0.09±0.02 in parity 4 for NBA, 0.07±0.02 in parity 2 to 0.16±0.02 in parity 3 for LTBW, 0.04±0.02 in parity 4 to 0.08±0.01 in parity 1 for NPW, and 0.16±0.02 in parity 1 to 0.20±0.02 in parity 2 for LTWW. Additive genetic correlation estimates from MTM were also variable, ranging from 0.29±0.24 between NBA in parity 1 and NBA in parity 2 to 0.99±0.05 between LTWW in parity 3 and LTWW in parity 4. Conclusion: The findings of this study highlight the advantage of using MTM for the genetic improvement of reproductive traits in swine and contribute to the development of sustainable swine breeding programs in Thailand.

• 방사성폐기물학회지
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• 제11권1호
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• pp.23-29
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• 2013

경제적이고 우수한 핵확산저항성을 갖는 파이로공정의 핵심 단위공정인 전해제련 공정에서 U와 TRU를 동시에 회수하기 위해 환원전극으로써 LCC가 사용된다. 한가지 원소만을 회수하는 금속음극과는 달리 LCC는 전기화학적으로 U와 TRU의 선택적 분리가 어려워 핵확산저항성을 높이는 기술의 핵심이라고 할 수 있다. LCC를 담아놓는 LCC 도가니는 U나 TRU로만 전착되어야하기 때문에 도가니는 전기적으로 절연되어야 한다. LCC와의 안정성과 회수된 TRU 및 용융염과의 화학적 안전성은 물론 공정 중 전착될 수 있는 금속 Li과의 반응성도 고려되어야하므로 LCC 도가니의 소재 특성은 매우 중요하다. 본 연구에서는 $Al_2O_3$, MgO, $Y_2O_3$, BeO 네 가지 대체 세라믹 소재의 화학적 안정성을 $500^{\circ}C$에서 모의 LCC로 열역학적 및 실험적으로 평가하였다. 세라믹 기판 위의 LCC 접촉각은 화학적 반응성을 예측하기 위해 시간에 따라 측정하였다. $Al_2O_3$는 가장 낮은 화학적 안정성 갖고 BeO는 재료 내에 존재하는 기공은 접촉각감소에 영향을 주었다. MgO, $Y_2O_3$는 우수한 화학적 안정성을 나타내었다.

• 한국식품저장유통학회지
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• 제23권4호
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• pp.553-559
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• 2016

고흥산과 캘리포나이산의 석류나무(Punica granatum L.) 과실을 과육, 종자 및 외과피로 분리하여 부위별 총 페놀성 함량, 안토시아닌 함량, DPPH 라디컬 소거능에 대한 항산화 활성 및 항암활성을 비교 검토하였다. 석류과실의 총 페놀성 함량은 외과피>주스>종자 순으로 유의적인 차이로 높게 나타났다. DPPH 라디컬 소거능은 외과피>주스>종자 순으로 높았고, 특히 외과피의 항산화 활성은 vitamin C와 유사한 활성을 보였다. 폐암세포주(Calu-6)에 대한 추출물의 항암활성은 외과피>종자>과육 순으로 높게 나타났다. 고흥산 석류 과실 부위별 총 안토시아닌 함량은 외과피>주스>종자 순으로 나타났고, 함유된 주요 안토시아닌은 Cy3,5G로서 가장 높은 함량을 보였고 그 다음이 Cy3G,Dp3,5G순으로 높게 나타났다. 총 페놀성 함량과 Calu-6 폐암세포주 활성 간이 $r^2=0.9706$으로 가장 높았고, 그 다음이 총 페놀성 함량과 DPPH 라디컬 소거능 간, DPPH 라디컬 소거능과 Calu-6 폐암세포주 활성 간 순으로 상관계수($r^2$)가 각각 0.8904, 0.8309로 나타나 상호 연관이 있음을 나타냈다.

• 한국식품저장유통학회지
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• 제8권4호
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• pp.481-487
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• 2001

본 연구에서는 다양한 기능성을 가지는 저령을 80% methanol로 추출한 후 그 농축액을 ehyl acetate와 water로 순차 분획하고, 각 분획물을 이용하여 인간유래 위암세포인 SNU668의 생장 저해 효과와 항산화 효과 및 각종 위장질환의 원인균으로 보고된 H. pylori에 대한 항균활성을 조사하였다. 인간유래의 위암세포인 SNU668에 대한 저해 효과는 methanol fraction 1 mg/$m\ell$의 농도에서 가장 강한 암세포 저해 활성을 보였고, water fraction 1 mg/$m\ell$의 농도에서는 43.9%의 저해 활성을 보였다. 그러나 ethyl acetate fraction에서는 23.7%로 비교적 낮은 저해율을 보였다. 반면에, 성인 남성의 정상 임파구는 분획종류에 관계없이 모두 90% 이상의 높은 생존율을 보였다. 한편, 저령 추출물의 항산화 효과는 생존을 75.2%를 보인 methanol 추출물이 43.7%의 생존율을 보인 water 분획물 보다 높았고, ethyl acetate 분획물은 항산화 효과 역시 가장 낮게 나타났다. H. pylori에 대한 저령의 항균활성은 1mg/disk의 농도에서 80% methanol 추출물 및 모든 분획물에서 H. priori에 대해 항균활성을 나타냈으며, 그중 water 분획물이 12mm의 inhibition zone을 형성하여 가장 우수한 항균활성을 보였다. Methanol 추출물과 ethyl acetate 분획물은 각각 11 및 9.5 mm의 inhibition zone을 형성하였다. H. pylori로부터 분리된 urease의 활성은 urea broth에서의흡광도와 pH의 변화를 조사한 결과, 23$^{\circ}C$에서 반응 1시간 동안 560 nm에서의 흡광도가 0에서 0.93까지 증가하였으며, pH는 7.0으로부터 8.1까지의 증가를 보였다. H.pylori로부터 분리된 urease의 활성은 80% methanol 추출물과 모든 분획물에 의해 억제되었으며, 각 분획물의 억제능은 항균실험 결과와 유사한 유형으로서 역시 10mg/$m\ell$의 농도에서 1시간 후 55%의 억제를 보인 water분획물의 억제능이 가장 우수했다.

• 생명과학회지
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• 제31권2호
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• pp.164-174
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• 2021

본 연구에서는 석류추출물의 기능성 활성 검증 및 화장품 개발을 위한 이용 가능성을 확인하였다. 전자공여능을 측정한 결과, 1,000 ㎍/ml의 농도에서 60.6%의 효과를 나타내었고, ABTS 소거능을 측정한 결과, 1,000 ug/ml의 농도에서 93.9%의 소거능을 보였다. 또한 elastase 억제 활성과 collagenase 억제 활성은 1,000 ㎍/ml의 농도에서 각각 30%와 47.2%의 억제 활성을 보였다. 석류추출물이 섬유아세포(CCD-986sk)의 증식에 미치는 영향을 조사하기 위해 MTT법으로 세포 생존성을 측정한 결과, 500 ㎍/ml 이하의 농도에서 130% 이상의 생존율이 확인됐다. 석류추출물을 처리한 섬유아세포에서 MMP-1, MMP-2, MMP-3의 단백질 발현은 100 ㎍/ml의 농도에서 각각 23.2%, 81.9%, 69.2%의 저해율을 나타내었다. 위 결과를 바탕으로 석류추출물이 0.1% (w/v)의 농도로 함유된 O/W 액정크림을 제조하여 4, 25, 45, 50℃에서 pH, 시간에 따른 변화, 온도별 안정성 등의 안정성을 조사한 결과, 모두 1개월간 안정성이 있는 것으로 확인되었으며 석류추출물의 화장품 소재로서의 가능성을 확인하였다.