• Biomolecules & Therapeutics
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• 제21권4호
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• pp.299-306
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• 2013

In the present study, we investigated the effect of ethanolic extract of the seed of Zizyphus jujuba var. spinosa (EEZS) on cholinergic blockade-induced memory impairment in mice. Male ICR mice were treated with EEZS. The behavioral tests were conducted using the passive avoidance, the Y-maze, and the Morris water maze tasks. EEZS (100 or 200 mg/kg, p.o.) significantly ameliorated the scopolamine-induced cognitive impairment in our present behavioral tasks without changes of locomotor activity. The ameliorating effect of EEZS on scopolamine-induced memory impairment was significantly reversed by a sub-effective dose of MK-801 (0.0125 mg/kg, s.c.). In addition, single administration of EEZS in normal naive mouse enhanced latency time in the passive avoidance task. Western blot analysis was employed to confirm the mechanism of memory-ameliorating effect of EEZS. Administration of EEZS (200 mg/kg) increased the level of memory-related signaling molecules, including phosphorylation of extracellular signal-regulated kinase or cAMP response element-binding protein in the hippocampal region. Also, the time-dependent expression level of brain-derived neurotrophic factor by the administration of EEZS was markedly increased from 3 to 9 h. These results suggest that EEZS has memory-ameliorating effect on scopolamine-induced cognitive impairment, which is mediated by the enhancement of the cholinergic neurotransmitter system, in part, via NMDA receptor signaling, and that EEZS would be useful agent against cognitive dysfunction such as Alzheimer's disease.

• Molecules and Cells
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• 제37권9호
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• pp.656-663
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• 2014

Gintonin, a novel, ginseng-derived G protein-coupled lysophosphatidic acid (LPA) receptor ligand, elicits $[Ca^{2+}]_i$ transients in neuronal and non-neuronal cells via pertussis toxin-sensitive and pertussis toxin-insensitive G proteins. The slowly activating delayed rectifier $K^+$ ($I_{Ks}$) channel is a cardiac $K^+$ channel composed of KCNQ1 and KCNE1 subunits. The C terminus of the KCNQ1 channel protein has two calmodulin-binding sites that are involved in regulating $I_{Ks}$ channels. In this study, we investigated the molecular mechanisms of gintonin-mediated activation of human $I_{Ks}$ channel activity by expressing human $I_{Ks}$ channels in Xenopus oocytes. We found that gintonin enhances $I_{Ks}$ channel currents in concentration- and voltage-dependent manners. The $EC_{50}$ for the $I_{Ks}$ channel was $0.05{\pm}0.01{\mu}g/ml$. Gintonin-mediated activation 1 of the $I_{Ks}$ channels was blocked by an LPA1/3 receptor antagonist, an active phospholipase C inhibitor, an $IP_3$ receptor antagonist, and the calcium chelator BAPTA. Gintonin-mediated activation of both the $I_{Ks}$ channel was also blocked by the calmodulin (CaM) blocker calmidazolium. Mutations in the KCNQ1 $[Ca^{2+}]_i$/CaM-binding IQ motif sites (S373P, W392R, or R539W)blocked the action of gintonin on $I_{Ks}$ channel. However, gintonin had no effect on hERG $K^+$ channel activity. These results show that gintonin-mediated enhancement of $I_{Ks}$ channel currents is achieved through binding of the $[Ca^{2+}]_i$/CaM complex to the C terminus of KCNQ1 subunit.

• Reproductive and Developmental Biology
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• 제33권4호
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• pp.257-262
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• 2009

This study was to investigate pregnancy rate of IVM/IVF/IVC Korean cattle (registered in government) embryos according to transport time course. For the production of embryos, oocytes recovered from slaughtered excellent grade cow and highly motile frozen-thawed bull semen (purchased from LIMC, KPN#497) was used. In vitro produced embryos were cultured in CR1aa medium for 8 days and some of them were frozen. The rate of average cleavage (>2-cell) was 83.0% (308/371) and blastocyst rate at day 8 was 34.7% (107/308). Among in vitro produced blastocyst embryos at day 8, most healthy embryos were freshly transferred on production day and some frozen embryos were direct transferred on appropriate day. These embryos were produced in a laboratory, embryo transfer (ET) was planned in 10 areas of the remote island (Jeju) from the laboratory by airplane. Thus, we examined the pregnancy rate in recipient cow according to embryo of transport time course before ET. From embryo transferred 44 recipient cows, overall pregnancy was 40.9% (18/44), these 18 cows were all calved [single, 94% (17/18); twin, 6% (1/18)] and total embryo implantation rate was 26% (19/66). Comparing transport time in the base of 6 hr, pregnancy rate in ET group required less 4 hr (60%, 9/15) was significantly higher than that required more 6 hr (26.3%, 5/19). In direct ET of freezing embryos, the pregnancy rate was 40% (4/10). However, it was difficult to find the meaning of temperature, pH and corpus luteum quality of recipients on comparison of pregnancy rate. When the cell death level of embryos according to storage time in thermos (straw container) before ET was measured by TUNEL staining, apoptotic index was increased with storage time-dependent. These results demonstrated that long distance transfer of IVM/IVF/IVC embryos is possible and the time of embryo transport is very important for the pregnancy rate on field trial.

• 한국식품과학회지
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• 제45권4호
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• pp.495-500
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• 2013

약용식물 3종에 대한 항균활성 결과, 보골지와 오미자 분획층에서 계혈등 분획층보다 높게 나타났다. 계혈등과 보골지 n-hexane 분획층에서 그람 음성균보다 그람 양성균에 대한 항균활성이 유의적으로 높게 나타났으며(p<0.05), 항균활성은 농도가 증가함에 따라 증가하였다. 계혈등 n-hexane 분획층의 L. monocytogenes에 대한 MIC는 7.765 (mg/mL), 보골지 n-hexane 분획층의 B. cereus, L. monocytogenes, E. coli 균에 대한 MIC는 2.03 (mg/mL), 오미자 분획층의 (L. monocytogenes), E. coli 균에 대한 MIC는 8.5 (mg/mL)로 나타났으며, 오미자 n-hexane 분획층의 MIC가 모든 균주에서 가장 낮게 나타났다. 약용식물 추출물의 항균활성에 대한 열 안정성 실험결과, L. monocytogens에 대한 계혈등 n-hexane 분획층을 제외하고는 $100^{\circ}C$에서 10분간의 열처리에도 항균 활성의 변화가 없었다. 본 연구 결과 보골지와 오미자 n-hexane 분획층에서 비교적 높은 항균활성을 나타냈다. 이러한, 약용 식물은 다양한 생리활성 및 기능성 성분을 포함하고 있어 천연 항균제의 유용한 자원으로 이용 될 수 있으며, 식품산업 발달에 기여 할 수 있을 것으로 사료된다.

• 한방재활의학과학회지
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• 제25권2호
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• pp.15-35
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• 2015

Objectives This study was performed to investigate the effects of Bujasasim-tang ethanol extract (BST) on oxidative stress, inflammation and osteoarthritic rat model. Methods To ensure safety of BST, heavy metal levels were measured and cytotoxicity test was done. In vitro, To evaluate antioxidative effects of BST, total phenolic contents, 1,1-diphenyl-2-picryl-hydrazyl (DPPH), 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) scavenging activity, reactive oxygen species (ROS) levels were measured. Also, to evaluate anti-inflammatory effects of BST treated group, total nitric oxide (NO) and pro-inflammatory cytokines (IL-$1{\beta}$, IL-6, TNF-${\alpha}$) levels were measured in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. In vivo, We injected MIA $50{\mu}l$ (60 mg/ml) into knee joints of rats to induce osteoarthritis. Rats were divided into total 3 groups (normal, control, BST treated group, each n=7). Normal group was not treated at all without inducing osteoarthritis and taken normal diet. Control group was induced osteoarthritis by MIA and taken with 2 ml of distilled water once a day for 4 weeks. BST treated group was induced osteoarthritis by MIA and taken BST 2 ml (200 mg/kg/mouse) once a day for 4 weeks. We evaluated dynamic weight bearing with the Incapacitance Test Meter. At the end of experiment, the rats were sacrificed to observe the functions of liver and kidney, changes of WBC, neutrophil, lymphocyte, monocyte levels in blood, to evaluate the levels of pro-inflammatory cytokines, tissue inhibitor of metallopreteinases-1 (TIMP-1), matrix metalloproteinase-9 (MMP-9), prostaglandin $E_2$ ($PGE_2$), leukotriene $B_4$ ($LTB_4$) within serum. We observed change of articular structures by Hematoxylin & Eosin (H&E), safranin-O staining method and measured amount of cartilage by micro CT-arthrography. Statistical analysis was done by unpaired student's t-test with significance level at p<0.05 in SPSS 11.0 for windows. Results 1. Safety of the BST was identified. 2. AST, ALT, BUN, creatinine levels of BST treated group were within normal limit. In vitro, 1. DPPH and ABTS free radical scavenging activities of BST showed dose-dependent increase. 2. ROS production were significantly decreased. 3. Total nitric oxide (NO) and IL-$1{\beta}$ production were decreased. 4. IL-6 and TNF-${\alpha}$ production were significantly decreased. In vivo, 1. Weight bearing ability was significantly increased. 2. WBC, neutrophil, lymphocyte, monocyte levels in blood were decreased. 3. IL-$1{\beta}$ and TNF-${\alpha}$ levels in serum were significantly decreased. and the IL-6 level was decreased. 4. TIMP-1, MMP-9, $LTB_4$, $PGE_2$ levels in serum were significantly decreased. 5. Cartilage volume of BST treated group was significantly increased. Also changes of cartilage, synovial membrane, fibrous tissue were suppressed. Conclusions The results obtained in this study Bujasasim-tang have effects of antioxidative, anti-inflammatory, relieve pain and protection of cartilage. Therefore we expect that Bujasasim-tang is effective treatment for osteoarthritis.

• 대한본초학회지
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• 제30권6호
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• pp.7-15
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• 2015

Objectives : Rosa laevigata Michx. has been used for the treatment of renal disease in traditional Korean medicine. In this study, we investigated cytoprotective effect of R. laevigata water extract (RLE) against oxidative stress induced by arachidonic acid (AA) + iron.Methods : To evaluate the protective effects of RLE against AA + iron-induced oxidative stress in HepG2 cell, cell viability and changes on apoptosis-related proteins were assessed by MTT and immunoblot analyses. The effects of RLE on reduced glutathione level, production of reactive oxygen species and mitochondrial membrane potential were also monitored. Furthermore, to verify underlying molecular mechanism, NF-E2-related factor 2 (Nrf2) was examined by immunoblot analysis. Additionally, Nrf2 transactivation and its downstream target genes expression were also determined by reporter gene and realtime RT-PCR analyses.Results : RLE pretreatment (30-300 μg/ml) prevented cells from AA + iron-mediated cell death in a concentration dependent manner. In addition, 100 μg/ml RLE inhibited AA + iron-induced glutathione depletion, reactive oxygen species production and mitochondrial dysfunction. RLE accumulated nuclear Nrf2 and also transactivated Nrf2, which was evidenced by antioxidant response element- and glutathione S-transferase A2-driven luciferase activities and mRNA level of glutamate-cysteine ligase catalytic subunit, NAD(P)H:quinone oxidoreductase 1 and sestrin 2. Moreover, protective effect of RLE against AA + iron was abolished in Nrf2 knockout cells.Conclusions : These results indicate that RLE has the ability to protect hepatocyte against oxidative stress through Nrf2 activation.

• 한국식품과학회지
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• 제37권4호
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• pp.597-602
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• 2005

인삼 유화 음료를 3가지 온도 조건에서 저장하면서 유화안정성, 당도, 진공도, pH, 산도, 색도(L, a, b값), 점도, 사포닌, 미생물 및 관능검사 등 총 10개 항목의 품질 요소들의 변화를 측정한 결과 저장기간 중 품질 변화가 크며 저장 온도에 따른 유의차를 나타내는 4가지 품질 지표인 유화 안정성, 산도, 점도, yellowness를 확인하였다. 품질 지표로 선정한 유화 안정성, 산도. 점도, yellowness의 온도 의존성을 Arrhenius식을 이용하여 판단한 결과, 점도의 경우 온도 의존성이 가장 큰 것으로 나타났으며, 유화 안정성의 경우 2단계로 변화가 일어나 저장 초기에 속도가 빠르고, 일정 기간 이후에 속도가 느렸다. 또한 산도 및 yellowness의 변화도 저장기간에 따라 변화하고 저장 온도에 따라 그 속도가 달라짐을 확인 할 수 있었다. 일반적으로 식품의 품질 변화의 온도 의존성을 나타내는 $Q_{10}$ value는 약 2의 값을 나타낸다고 하였으나, 본 연구에서 품질 지표로 선정된 4가지 요소의 $Q_{10}$ value는 각각 다른 값을 나타내었다. 이로 보아 같은 식품이라도 품질 인자에 따라 각기 다른 $Q_{10}$ value를 가지는 것을 알 수 있었으며, 일률적으로 $Q_{10}$ value를 적용하는 것은 적절하지 않음을 알 수 있었다.

• 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.

• 산업식품공학
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• 제15권4호
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• pp.332-337
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• 2011

치환제인 PO 농도(2%, 6%, 12%)를 달리하여 제조한 하이드록시프로필화 쌀전분에 가교제인 $POCl_{3}$ 농도(0.005%, 0.02%)를 달리하여 처리한 HPCL 쌀전분을 제조하고, HPCL 쌀전분의 용해도, 팽윤력, pasting 특성 및 열적 특성을 연구하였다. 팽윤력은 일반 쌀전분 보다 다소 낮은 온도에서 증가되기 시작하였으나 그 이후는 모두 낮은 수치를 보여 HPCL 쌀전분이 일반 쌀전분보다 완만한 상승을 나타내었다. 용해도는 HPCL 쌀전분이 일반 쌀전분보다 전체적으로 낮은 값을 나타내었다. 한편 PO 함량이 증가할수록 팽윤력과 용해도 모두 증가하는 경향을 나타내었다. RVA pasting분석 결과, HPCL 쌀전분의 경우 일반 쌀전분보다 최고 점도는 낮아졌고 holding strength와 finalviscosity는 높은 경향을 나타내었으며, breakdown은 낮아지고 setback은 높은 것으로 나타났다. 열적 특성 분석 결과 HPCL 쌀전분의 경우 일반 쌀 전분보다 낮은 $T_{o}, T_{p}, T_{c}$ 및 ${\Delta}H$를 나타내었으며, $POCl_{3}$ 함량이 적을수록 감소폭이 더 큰 것으로 나타났다. 따라서 HPCL 쌀전분의 경우 RVA pasting 특성은 PO 첨가량, 즉 하이드록시프로필화에 따른 영향이 크다고 볼 수 있고 DSC 열적특성은 하이드록시프로필화 및 가교화 모두 영향을 미치는 것으로 생각된다.

• 한국식품과학회지
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• 제53권1호
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• pp.34-39
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• 2021

본 연구에서는 천연유래 물질로부터 apoptosis를 유도하여 항암 활성이 있는지에 관한 실험으로 사람 난소암 세포주인 OVCAR-3 세포에 함초의 유기용매별 추출물을 처리하여 결과를 확인하였다. MTS 측정으로 세포 생존율을 확인한 결과 DCM 분획물에서 농도별로 유의적인 세포수의 감소를 보였으며, annexin V/FITC-PI 염색에 의해 apoptosis 유도로 세포가 사멸함을 확인하였다. DCM 분획물 처리는 세포주기에서 Sub-G1기의 증가로 세포증식이 억제됨을 보여준다. 세포의 내인성 경로에 관여하는 Bcl-2 family에 속하는 Bax, Bak, Bcl-2, Bcl-xL의 상호작용을 mRNA 수준에서 확인한 결과, DCM 분획물처리에서 Bax, Bak가 증가하고, Bcl-2의 감소를 동반하여 세포사멸의 신호전달 경로가 진행됨을 확인할 수 있었다. 본 연구 결과를 바탕으로 함초를 이용하여 여성의 난소암에 예방적 기능성 식품을 개발할 수 있을 것으로 판단되며, 함초의 DCM 분획물에 대해 깊이 있는 심층적 연구가 요구된다.