• Asian-Australasian Journal of Animal Sciences
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• 제27권5호
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• pp.733-742
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• 2014

The glucagon-like peptide 2 (GLP-2) that is expressed in intestine epithelial cells of mammals, is important for intestinal barrier function and regulation of tight junction (TJ) proteins. However, there is little known about the intracellular mechanisms of GLP-2 in the regulation of TJ proteins in piglets' intestinal epithelial cells. The purpose of this study is to test the hypothesis that GLP-2 regulates the expressions of TJ proteins in the mitogen-activated protein kinase (MAPK) signaling pathway in piglets' intestinal epithelial cells. The jejunal tissues were cultured in a Dulbecco's modified Eagle's medium/high glucose medium containing supplemental 0 to 100 nmol/L GLP-2. At 72 h after the treatment with the appropriate concentrations of GLP-2, the mRNA and protein expressions of zonula occludens-1 (ZO-1), occludin and claudin-1 were increased (p<0.05). U0126, an MAPK kinase inhibitor, prevented the mRNA and protein expressions of ZO-1, occludin, claudin-1 increase induced by GLP-2 (p<0.05). In conclusion, these results indicated that GLP-2 could improve the expression of TJ proteins in weaned pigs' jejunal epithelium, and the underlying mechanism may due to the MAPK signaling pathway.

• 대한본초학회지
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• 제36권3호
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• pp.1-8
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• 2021

Objective : Reflux esophagitis (RE), one of gastroesophageal reflux disease (GERD), is a disease that causes inflammation due to reflux of stomach contents such as stomach acid and pepsin due to the unstable gastroesophageal sphincter, and is currently increasing worldwide. The currently used treatment for reflux esophagitis has various side effects. Therefore, in this study the effect of Toosendan Fructus extract on chronic acid reflux esophagitis in rats was evaluated in order to find a new treatment material for reflux treatment. Methods : After inducing reflux esophagitis through surgery, the group was separated and the drug was administered for 2 weeks; Normal rats (Normal, n=8), chronic acid reflux esophagitis rats (Control, n=8), Toosendan Fructus 200 mg/kg body weight/day-treated chronic acid reflux esophagitis rats (TF, n=8). After, we were taken esophageal tissue and esophageal mucosa damage was identified, and analyzed the expression of NADPH oxidase, AP-1/MAPK-related proteins, and tight junction proteins by western blot in esophageal tissue. Results : Toosendan Fructus administration significantly protected the esophageal mucosal damage of reflux esophagitis. Also, Toosendan Fructus significantly reduced the expression of NADPH oxidases (NOX2 and p22^phox) and AP-1/MAPK-related proteins (c-Fos, c-Jun, p-p38, p-ERK, and p-JNK). In addition, it significantly increased the expression of tight junction proteins (Occludin, Claudin-3, and Claudin-4). Conclusions : These results suggest that Toosendan Fructus reduced damage to the esophageal mucosa by protecting the esophageal mucosa by upregulating tight junctions proteins as well as inhibiting the AP-1/MAPK pathway through reducing NADPH oxidases expression.

• Journal of Animal Science and Technology
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• 제64권3호
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• pp.500-514
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• 2022

The blood-epididymis barrier (BEB) forms a unique microenvironment that is crucial for the maturation, protection, transport, and storage of spermatozoa in the epididymis. To characterize the function of tight junctions (TJs), which are constitutive components of the BEB, we determined the expression and localization of TJ proteins such as zonula occludens (ZO)-1, 2, and 3, occludin, and claudin3 (Cldn3) during postnatal development in the goat epididymis. To assess the expression patterns of TJ proteins in immature (3 months of age) and mature (14 months of age) goat epididymides, two different experimental methods were used including immunofluorescence labeling and western blotting. We show that, ZO-1, 2, and 3, and occludin, were strictly expressed and localized to the TJs of the goat epididymis, whereas Cldn3 was present in basolateral membranes as well as TJs. All TJ proteins examined were more highly expressed in the immature epididymis compared to levels in mature tissue. In conclusion, our study indicates that at least five TJ proteins, namely ZO-1, ZO-2, ZO-3, occludin, and Cldn3, are present in TJs, and the expression strength and pattern of TJ proteins tend to be age dependent in the goat epididymis. Together, these data suggest that the distinct expression patterns of TJ proteins are essential for regulating components of the luminal contents in the epididymal epithelium and for forming adequate luminal conditions that are necessary for the maturation, protection, transport, and storage of spermatozoa in the goat epididymis.

• 대한한방내과학회지
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• 제37권1호
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• pp.16-25
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• 2016

Objectives: Gilgyung-tang (GGT) has been used as one of the main multi-herb formulas to treat “Peo-ong” (lung abscess). In this study, we investigated the inhibitory effects of water extracts of GGT on cell migration and invasion, two critical cellular processes that are often deregulated during metastasis, in human bladder cancer 5637 cells.Methods: Effects on cell viability were quantified using an MTT assay. To analyze the anti-metastatic effects, we conducted a wound healing migration assay, an in vitro invasiveness assay, and a measurement of the transepithelial electrical resistance (TER). The expression of protein and mRNA were measured by Western blotting and real-time polymerase chain reaction (RT-PCR), respectively.Results: GGT markedly inhibited the cell motility and invasiveness of 5637 cells within the concentration range that was not cytotoxic. The inhibitory effects of GGT on cell invasiveness were associated with tightening of the tight junctions (TJs), which was demonstrated by an increase in the TER. The RT-PCR and Western blotting results indicated that GGT decreased the levels of claudin proteins. GGT also inhibited the activity and expression of matrix metalloproteinase (MMP)-2 and -9 and simultaneously increased the levels of tissue inhibitor of metalloproteinase-1 and -2.Conclusions: Our findings suggest that GGT reduces both the migration and the invasion of 5637 cells by modulating the activity of TJs and MMPs.

• 대한한방소아과학회지
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• 제35권3호
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• pp.118-127
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• 2021

Objective The purpose of this study was to confirm the effect of Scutellaria baicalensis extract on skin damage recovery and inflammation relief in atopic dermatitis-induced mice through Endocannabinoid system (ECS) control. Methods 6-week-old Balb/C mice were divided into control group (Ctrl), atopic dermatitis induced group (ADE), palmitoylethanolamide (PEA) administered group after atopic dermatitis induced (PEAT), and Scutellaria baicalensis extract administered group after atopic dermatitis induced (SBT). Seven animals were assigned for each group. After drug administration for 3 weeks after inducing atopic dermatitis, Claudin and 8-OHdG were observed to confirm the recovery of the skin damage in each group. To confirm ECS regulation, CB1, CB2, and GPR55 were observed. To confirm the anti-inflammatory effect, Fc ε receptor, and MMP-9 was observed. Results Claudin positive reaction was significantly increased in SBT compared to ADE and PEAT. 8-OHdG positive reaction was significantly decreased in SBT compared to ADE and PEAT. CB1, CB2, and GPR55 positive responses were significantly increased in SBT compared to ADE and PEAT. Fc ε receptor and MMP-9 positivity were significantly decreased in SBT compared to ADE and PEAT. Conclusion It was confirmed that the Scutellaria baicalensis extract can reduce the inflammation of atopic dermatitis by restoring the structural damage of the skin lipid barrier through ECS activity.

• 한국식품과학회지
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• 제53권2호
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• pp.165-173
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• 2021

본 연구에서는 UPM에 의해 유발되는 NCI-H441 세포 내 ROS 생성과 그에 따른 tight junction 단백질(N-cadherin, fibronectin, occluding, ZO-1, 및 claudin-4 등)의 발현 억제를 확인하였고 ECL의 전처리로 인하여 해당 ROS 생성 및 tight junction 단백질 발현 억제 현상이 완화되는 보호효과를 확인하였다. 그 결과, 폐 세포에서 UPM에 의해 유발된 산화 스트레스 및 tight junction 단백질의 발현 감소가 ECL을 선 처리함으로써 억제되어 결과적으로 산화 스트레스와 같은 미세먼지에 의한 폐 손상으로부터 보호 효과를 나타내고 tight junction 수준의 유지를 확인하였다. 이러한 결과를 통해 ECL 추출물은 폐와 기관지의 보호효과를 나타내며 폐 관련 질병의 예방효과가 있는 기능성 천연 물질로써 활용이 가능할 것으로 기대된다.

• 한국유기농업학회지
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• 제29권1호
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• pp.85-96
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• 2021

본 결과를 통해, 왕우렁이 추출물의 역류성 식도염 모델에서 위 내용물의 역류로 인한 조직손상에 대한 보호효과를 확인하였다. 식도 점막의 육안 소견과 조직병리학적 분석 결과 점막의 괴사, 염증세포의 침윤 등의 증상이 완화됨을 확인하였으며, 상피조직의 tight junction을 보호하여 위산과 펩신으로부터 점막을 보호하는 것을 확인하였다. 이러한 왕우렁이 추출물의 식도 보호효과는 NF-κB 기전의 조절을 통해 점막의 염증반응을 억제하여 나타나는 것으로 생각된다. 이상의 결과를 종합하였을 때, 왕우렁이 추출물이 역류성 식도염 완화를 위한 기능성 소재로서의 가능성을 가지는 것으로 사료된다.

• Journal of Ginseng Research
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• 제46권3호
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• pp.464-472
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• 2022

Background: Gut microbiota influence the central nervous system through gut-brain-axis. They also affect the neurological disorders. Gut microbiota differs in patients with Alzheimer's disease (AD), as a potential factor that leads to progression of AD. Oral intake of Korean Red Ginseng (KRG) improves the cognitive functions. Therefore, it can be proposed that KRG affect the microbiota on the gut-brain-axis to the brain. Methods: Tg2576 were used for the experimental model of AD. They were divided into four groups: wild type (n = 6), AD mice (n = 6), AD mice with 30 mg/kg/day (n = 6) or 100 mg/kg/day (n = 6) of KRG. Following two weeks, changes in gut microbiota were analyzed by Illumina HiSeq4000 platform 16S gene sequencing. Microglial activation were evaluated by quantitative Western blot analyses of Iba-1 protein. Claudin-5, occludin, laminin and CD13 assay were conducted for Blood-brain barrier (BBB) integrity. Amyloid beta (Aβ) accumulation demonstrated through Aβ 42/40 ratio was accessed by ELISA, and cognition were monitored by Novel object location test. Results: KRG improved the cognitive behavior of mice (30 mg/kg/day p < 0.05; 100 mg/kg/day p < 0.01), and decreased Aβ 42/40 ratio (p < 0.01) indicating reduced Aβ accumulation. Increased Iba-1 (p < 0.001) for reduced microglial activation, and upregulation of Claudin-5 (p < 0.05) for decreased BBB permeability were shown. In particular, diversity of gut microbiota was altered (30 mg/kg/day q-value<0.05), showing increased population of Lactobacillus species. (30 mg/kg/day 411%; 100 mg/kg/day 1040%). Conclusions: KRG administration showed the Lactobacillus dominance in the gut microbiota. Improvement of AD pathology by KRG can be medicated through gut-brain axis in mice model of AD.

• 대한화장품학회지
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• 제50권2호
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• pp.179-192
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• 2024

본 연구에서는 천연물 6 종 앵두, 으름, 살구, 야콘, 금귤 및 핑거라임을 활용하여 스트레스 완화에 의한 수면 개선 및 피부장벽 개선 효과를 검증하였다. 실험 결과, 6 종의 천연물 중 앵두, 핑거라임, 으름 및 금귤이 유의적으로 스트레스 호르몬인 cortisol의 생성을 억제하였다. 또한, 수면 조절에 관여하는 GABA 생성 효소인 GAD67의 발현은 앵두 정제수 추출물 및 핑거라임 50% 에탄올 추출물에서 유의한 증가 효과를 보였으며 앵두 용매별 추출물은 총 폴리페놀의 함량이 가장 높게 확인되었다. 스크리닝 결과 활성이 가장 높은 앵두 추출물을 활용하여 후속 실험을 진행하였다. 앵두 용매별 추출물 중, 30% 에탄올 추출물이 가장 높은 DPPH radical 소거 활성을 보였으며, 정제수 추출물은 GABA 생성 및 피부 장벽 인자인 filaggrin과 claudin-1의 발현을 가장 높게 증가시켰다. HPLC 분석을 통해 앵두 추출물의 주성분으로 quercitrin이 확인되었고 추출 용매별 quercitrin 함량은 30% 에탄올 > 정제수 > 70% 에탄올 > 50% 에탄올의 추출 용매 순으로 높은 함량을 보였다. Quercitrin은 cortisol의 생성을 농도의존적으로 억제하였으며, cortisol에 의해 감소되었던 GAD67 발현 및 GABA 생성을 유의적으로 증가시켰다. 연구를 통해 앵두 추출물이 스트레스를 완화시켜 수면 개선 및 피부 장벽 강화에 도움을 주는 화장품 소재로서 활용이 가능함을 입증하였다.

• Animal Bioscience
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• 제36권9호
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• pp.1403-1413
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• 2023

Objective: Intestinal alkaline phosphatase (IAP) maintains intestinal homeostasis by detoxifying bacterial endotoxins and regulating gut microbiota, and lipid absorption. Antibiotics administered to animals can cause gut dysbiosis and barrier disruption affecting animal health. Therefore, the present study sought to investigate the role of IAP in the intestinal environment in dysbiosis. Methods: Young male mice aged 9 weeks were administered a high dose of antibiotics to induce dysbiosis. They were then sacrificed after 4 weeks to collect the serum and intestinal organs. The IAP activity in the ileum and the level of cytokines in the serum samples were measured. Quantitative real-time polymerase chain reaction analysis of RNA from the intestinal samples was performed using primers for tight junction proteins (TJPs) and proinflammatory cytokines. The relative intensity of IAP and toll-like receptor 4 (TLR4) in intestinal samples was evaluated by western blotting. Results: The IAP activity was significantly lower in the ileum samples of the dysbiosis-induced group compared to the control. The interleukin-1 beta, interleukin-6, and tumor necrosis factor-alpha concentrations were significantly higher in the ileum samples of the dysbiosis-induced group. The RNA expression levels of TJP2, claudin-3, and claudin-11 showed significantly lower values in the intestinal samples from the dysbiosis-induced mice. Results from western blotting revealed that the intensity of IAP expression was significantly lower in the ileum samples of the dysbiosis-induced group, while the intensity of TLR4 expression was significantly higher compared to that of the control group without dysbiosis. Conclusion: The IAP activity and relative mRNA expression of the TJPs decreased, while the levels of proinflammatory cytokines increased, which can affect intestinal integrity and the function of the intestinal epithelial cells. This suggests that IAP is involved in mediating the intestinal environment in dysbiosis induced by antibiotics and is an enzyme that can potentially be used to maintain the intestinal environment in animal health care.