• 한국식품과학회지
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• 제42권3호
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• pp.335-342
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• 2010

본 연구에서는 홍삼성분에 의한 장상피세포 간극(tight junction)의 안정화에 대해서 조사하였다. Transwell에 배양한 Caco-2 단세포층을 HBSS로 치환한 후 단세포층의 상층부에 홍삼추출물을 처리하고 96시간동안 배양하면서 TEER(transepithelial electrical resistance)수치의 변화를 조사하였다. 홍삼 추출물을 $20\;{\mu}g/mL$ 이하의 농도로 상층부에 첨가하였을 때에는 시간이 지남에 따라 점차 감소하는 대조구와는 달리 TEER수치가 덜 감소하였다. 한편 $130\;{\mu}g/mL$ 이상의 농도로 처리하였을 때는 TEER수치가 현저히 감소하였으나 MTT assay 결과 세포독성에 의한 것은 아니었다. 인삼으로부터 분리한 저분자물질, 다당류, 단백질, 사포닌, 폴리페놀 조획분을 각각 상층부에 첨가하였을 때, 장상피세포 간극의 안정화 활성을 나타내는 성분은 폴리페놀 및 사포닌 조획분이었으며, 전자는 높은 활성을 후자는 낮은 활성을 나타내었다. 이처럼 홍삼추출물의 폴리페놀 조획분과 사포닌 조획분은 장상피세포의 간극을 안정화시킴으로서, 식품 알레르겐의 체내유입을 차단하여 알레르기억제에 기여할 수 있을 것으로 추측된다.

• Journal of Chest Surgery
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• 제54권6호
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• pp.449-453
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• 2021

The 2020 American College of Cardiology focused update on the mitral regurgitation (MR) pathway provides an excellent summary of the decision-making trees in the treatment of severe MR, in which 2 main branches of the flowchart are suggested depending on whether MR is primary or secondary. Surgery is suggested as preferable over transcatheter edge-to-edge repair (TEER) in primary MR that needs intervention. The decision-making for secondary MR generally prioritizes TEER over surgery according to the guidelines, but further stratification is necessary based on the pathophysiologic mechanisms of MR. TEER is probably the more suitable option in secondary MR caused by left ventricular dysfunction or dilatation, given the high perceived surgical risks, despite the lack of sufficient evidence in support of overt clinical benefits from surgical therapy in these patients. In atrial functional MR associated with atrial fibrillation (AF), however, concomitant ablation of AF seems to be a desirable option, as it has been demonstrated to be a key factor leading to improved survival, reduced stroke risk, and more durable mitral and tricuspid function in patients undergoing mitral surgery. Therefore, atrial functional MR requiring intervention may be best treated by surgical therapy that combines mitral repair and AF ablation in the majority of patients. This particular issue, however, needs further research to obtain scientific evidence to guide optimal management strategies.

• 생명과학회지
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• 제26권9호
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• pp.1082-1087
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• 2016

본 연구에서는 염증성 사이토카인 IL-1α가 소장의 tight junction (TJ)의 integrity에 미치는 영향을 평가하고 항염증 효능이 있다고 알려진 curcumin (CCM)이 IL-1α에 의한 TJ 손상 예방효과를 알아보고자 Caco-2 세포 모델을 이용하여 실험하였다. Caco-2 세포 단층의 TJ integrity에 대한 IL-1α의 영향을 FITC-dextran flux와 transepithelial electrical resistance (TEER)를 측정하여 검증하였다. IL-1α를 100 ng/ml의 농도로 Caco-2 세포 단층의 상부에 24시간 동안 처리하였을 때 처리 2시간 이후 FITC-dextran의 flux가 유의적으로 증가하였고, IL-1α 처리시간에 비례하여 상승하였다. 또한 IL-1α의 처리는 TEER 값을 유의적으로 감소시켜 IL-1α에 의한 TJ 손상을 확인할 수 있었다. 반면 CCM의 전처리는 이러한 IL-1α에 의한 TJ 기능 저하를 거의 완전히 예방하는 것을 알 수 있었다. 이상의 결과로 염증성 사이토카인 IL-1α이 TJ의 integrity 조절에 부정적인 영향을 미치며 이는 강황에서 발견되는 CCM 전처리에 의해 효과적으로 억제될 수 있음을 보여준다. 본 연구 결과와 관련되어 TJ 구성단백질 발현 및 작용 기작에 대한 분자세포생물학적 연구와 in vivo 연구가 필요하다.

• 대한화장품학회지
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• 제45권4호
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• pp.389-397
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• 2019

리소좀(lysosome)은 진핵세포에서 에너지 대사 및 세포 내 소화 작용에 관여하는 세포 소기관으로 protease, nuclease, glycosidase, lipase, phosphatase 들이 다수 존재한다. 우리는 선행 연구결과들을 통해 난백 리소좀의 멜라닌 색소 탈색능을 보고하였다[8]. 그러나 B16F10 melanocyte 세포주에서 난백 리소좀에 의한 멜라닌 함량 변화 및 피부장벽 조절 연구는 거의 보고되지 않았다. 따라서 우리는 계란 난백(egg white)으로부터 추출한 lysosome-related organelle extract (LOE)에 의한 세포 내 멜라닌 함량 변화 및 피부장벽 강화 효과를 규명하고자 하였다. 먼저 LOE의 미백 효능을 확인하기 위해 B16F10 세포주를 이용하여 세포독성 평가를 진행하였다. B16F10 세포주에서 LOE에 의한 세포독성은 0에서 20 mg/mL 농도에서 관찰되지 않았으나, 40 mg/mL 부터 세포독성이 관찰되어 이후 모든 실험에서 최대 농도값을 20 mg/mL로 설정하였다. 먼저 LOE를 이용한 melanin contents assay 결과, 음성 대조군인 α-MSH 처리군 대비 LOE 처리군 5, 10, 20 mg/mL 농도에서 61.5 ± 4.0%, 61.4 ± 7.3%, 58.3 ± 8.3%로 세포 내 멜라닌 함량이 감소되는 것을 확인하였고, 20 mg/mL 농도 조건에서 MITF 발현 억제도 관찰하였다. LOE의 피부 장벽에 미치는 영향을 관찰하기 위해 각질형성세포주(HaCaT)를 이용하여 TEER (trans-epithelial electrical resistance) assay를 수행한 결과, LOE에 의해 농도 의존적으로 TEER 저항값이 증가하여 LOE가 피부장벽 강화에도 효과가 있음을 알 수 있었다. 또한 피부 염증 유발을 위한 TNF-α 처리조건에서도 LOE는 TEER 저항값을 증가시켜 염증 유발 조건에서도 LOE에 의해 피부장벽이 정상적으로 회복되었음을 알 수 있었다. 마지막으로 cell migration assay를 통해 LOE에 의한 세포이동 촉진 효과를 관찰한 결과, LOE는 세포분열 및 세포이동을 촉진시켰다. 위 결과들을 통해 LOE는 미백 기능 뿐 아니라 피부재생 및 피부장벽 강화에도 효과를 나타내는 소재이며, 효소안정화 및 제형화 기술이 접목된다면 향후 새로운 미백 기능성 화장품 소재로도 개발될 수 있을 것이다.

• Journal of Pharmaceutical Investigation
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• 제32권1호
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• pp.21-26
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• 2002

The primary culture of human nasal epithelial cell monolayer was performed on a Transwell. The effect of various factors on the tight junction formation was observed in order to develop an in vitro experimental system for nasal transport studies. Human nasal epithelial cells, collected from human normal inferior turbinates, were plated onto diverse inserts. After 4 days, media of the apical surface was removed for air-liquid interface (ALI) culture. Morphological characteristics was observed by transmission electron microscopy (TEM). A polyester membrane of $0.4\;{\mu}m$ pore size was determined as the most effective insert based on the change in the transepithelial electric resistance (TEER) value as well as the $^{14}C-mannitol$ transport study. The ALI method was effective in developing the tight junction as observed in the further increase in the TEER value and reduction in the permeability coefficient $(P_{app})$ of $^{14}C-mannitol$ transport. Results of the transport study of a model drug, budesonide, showed that the primary culture system developed in this study could be further developed and applied for in vitro nasal transport studies.

• Journal of Pharmaceutical Investigation
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• 제34권6호
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• pp.483-489
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• 2004

Fexofenadine HCl is non-sedating histamine H1 receptor antagonist that can be used for the treatment of seasonal allergic rhinitis. The objective of this study was to investigate whether the carriers of deformable liposomes can enhance the transepithelial permeability of fexofenadine HCl across the in vitro ALI human nasal monolayer model. Characterization of this model was achieved by bioelectric measurements and morphological studies. The passage 2 and 3 of cell monolayers exhibited the TEER value of $2852\;{\pm}\;482\;ohm\;{\times}\;cm^2$ on 11 days of seeding and maintained high TEER value for 5 days. The deformable liposome of fexofenadine HCl was prepared with phosphatidylcholine (PC) and cholic acid using extruder method. The mean particle size was about 200 nm and the maximum entrapment efficiency of 33.0% was obtained in the formulation of 1% PC and $100\;{\mu}g/ml$ fexofenadine HCl. The toxicity of the deformable liposome to human nasal monolayers was evaluated by MTT assay and TEER value change. MTT assay showed that it has no toxic effect on the nasal epithelial cells in 2-hour incubation when the PC concentration was below 1%. However, deformable liposome could not enhance the transepithelial permeability $(P_{app})$ and cellular uptake of fexofenadine HCl. In conclusion, the in vitro model could be used in nasal drug transport studies and evaluation of transepithelial permeability of formulations.

• 대한한의학방제학회지
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• 제32권2호
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• pp.121-128
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• 2024

Objectives : This study aimed to investigate the protective effects of mixed extracts from Scutellaria baicalensis Georgi, Alisma orientale Juzepzuk, and Atractylodes japonica Koidzumi on interleukin (IL)-6-induced damage to tight junction (TJ) integrity in a Caco-2 cell model. Methods : We assessed the TJ integrity of Caco-2 monolayers by measuring the flux of FITC-labeled dextran and transepithelial electrical resistance (TEER). Additionally, we evaluated the expression of TJ proteins, such as ZO-1 and Occludin. Results : Treatment with IL-6 (50 ng/ml) increased TJ permeability and decreased TEER values of Caco-2 monolayers. Pretreatment with HTB (50-200 ㎍/ml) for 1 h significantly alleviated IL-6-induced TJ disruption, as evidenced by reduced TJ permeability and increased TEER values. Furthermore, HTB reversed the IL-6-induced inhibition of TJ protein expression, including ZO-1 and Occludin. Conclusions : These findings indicate that HTB protects barrier function by reversing the IL-6-induced decrease in TJ integrity and the suppression of TJ protein expression.

• Journal of Animal Science and Technology
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• 제65권5호
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• pp.1040-1052
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• 2023

The objective of this study was to investigate the protective effects of functional nutrients including various functional amino acids, vitamins, and minerals on chicken intestinal epithelial cells (cIECs) treated with oxidative stress. The cIECs were isolated from specific pathogen free eggs. Cells were exposed to 0 mM supplement (control), 20 mM threonine (Thr), 0.4 mM tryptophan (Trp), 1 mM glycine (Gly), 10 μM vitamin C (VC), 40 μM vitamin E (VE), 5 μM vitamin A (VA), 34 μM chromium (Cr), 0.42 μM selenium (Se), and 50 μM zinc (Zn) for 24 h with 6 replicates for each treatment. After 24 h, cells were further incubated with fresh culture medium (positive control, PC) or 1 mM H₂O₂ with different supplements (negative control, NC and each treatment). Oxidative stress was measured by cell proliferation, whereas tight junction barrier function was analyzed by fluorescein isothiocyanate (FITC)-dextran permeability and transepithelial electrical resistance (TEER). Results indicated that cell viability and TEER values were less (p < 0.05) in NC treatments with oxidative stress than in PC treatments. In addition, FITC-dextran values were greater (p < 0.05) in NC treatments with oxidative stress than in PC treatments. The supplementations of Thr, Trp, Gly, VC, and VE in cells treated with H₂O₂ showed greater (p < 0.05) cell viability than the supplementation of VA, Cr, Se, and Zn. The supplementations of Trp, Gly, VC, and Se in cells treated with H₂O₂ showed the least (p < 0.05) cellular permeability. In addition, the supplementation of Thr, VE, VA, Cr, and Zn in cells treated with H₂O₂ decreased (p < 0.05) cellular permeability. At 48 h, the supplementations of Thr, Trp, and Gly in cells treated with H₂O₂ showed the greatest (p < 0.05) TEER values among all treatments, and the supplementations of VC and VE in cells treated with H₂O₂ showed greater (p < 0.05) TEER values than the supplementations of VA, Cr, Se, and Zn in cells treated with H₂O₂. In conclusion, Thr, Trp, Gly, and VC supplements were effective in improving cell viability and intestinal barrier function of cIECs exposed to oxidative stress.

• Archives of Pharmacal Research
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• 제26권12호
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• pp.1102-1108
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• 2003

We studied the effects of phytolaccosides, saponins from Phytolacca americana, on the intestinal absorption of heparin in vitro and in vivo. The absorption enhancing activity of these compounds (phytolaccosides B, $D_2$, E, F, G and I) was determined by changes in transepithelial electrical resistance (TEER) and the transport amount of heparin disaccharide, the major repeating unit of heparin, across Caco-2 cell monolayers. With the exception of phytolaccoside G, all of them decreased TEER values and increased the permeability in a dose-dependent and time-dependent manner. In vitro, phytolaccosides B,$D_2$, and E showed significant absorption enhancing activities, while effects by phytolaccoside F and I were mild. In vivo, phytolaccoside E increased the activated partial thromboplastin time (APTT) and thrombin time, indicating that phytolaccoside E modulated the transport of heparin in intestinal route. Our results suggest that a series of phytolaccosides from Phytolacca americana can be applied as pharmaceutical excipients to improve the permeability of macromolecules and hydrophilic drugs having difficulty in absorption across the intestinal epithelium.

• Archives of Pharmacal Research
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• 제25권1호
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• pp.86-92
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• 2002

The enhancement of paracellular transport of heparin disaccharide using several absorption enhancers across Caco-2 cell monolayers was tested . The cytotoxicity of these enhancers was also examined. The enhancing effects by Quillaja saponin, diponin glycyrrhizinate, $18{\beta}-glycyrrhetinic$ acid, sodium caprate and taurine were determined by changes in transepithelial electrical resistance (TEER) and the amount of heparin disaccharide transported across Caco-2 cell monolayers. Among the absorption enhancers, $18{\beta}-glycyrrhetinic$ acid arid taurine decreased TEER and increased the permeability of heparin disaccharide in a dose-dependent and time-dependent manner with little or negligible cytotoxicity. Our results indicate that these absorption enhancers can widen the tight junction, which is a dominant paracellular absorption route of hydrophilic compounds . It is highly possible that these absorption enhancers can be applied as pharmaceutical excipients to improve the transport of macromolecules and hydrophilic drugs having difficulty in permeability across the intestinal epithelium.