• 폴리머
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• 제38권1호
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• pp.54-61
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• 2014

Bombyx mori silk fibroin(SF)과 블렌드 필름을 만들기 위하여 키토산에 1,3-propanesultone을 반응시켜 수용성 sulfobetaine chitosan(SCs)을 제조하였다. 여러 가지 비율의 SF/SCs 블렌드 필름을 B. mori SF와 SCs의 수용액을 혼합하여 제조하였다. 수용액으로부터 얻어진 SF/SCs 블렌드 필름의 구조와 형태 변화는 분광학적 및 열적 분석을 통해 규명하였다. SF와 SCs의 혼합 비율에 따른 인공 피부나 화상치료 목적의 비이오재료로서의 물리적 및 기계적 성질에 미치는 영향을 조사하였다. X-선 분석으로 두 생체고분자 사이에 좋은 친화성을 보여주고 있음을 알 수 있었으며 기계적 성질도 SCs의 함량이 증가하면 크게 증가하였다. $37^{\circ}C$에서 phosphate buffered saline solution 용액 중에서 in vitro 분해 실험을 8주 동안 시행한 결과 46.4%가 분해됨을 알 수 있었다. MC3T3-E1 세포에 의한 독성 실험 결과 무독성을 나타내 주었으며, 3일의 배양 후 SF/SCs 필름의 상대 세포 수는 최적화된 tissue culture plastic보다 약간 낮게 나타남을 알 수 있었다.

• 동의생리병리학회지
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• 제33권2호
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• pp.131-140
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• 2019

The objective of present study is to evaluate concentration-dependent in vitro anti-osteoarthritic (OA) effects of synergic mixed formula consisted of dried pomegranate juice concentrate powder, Eucommiae Cortex aqueous extract and Achyranthis Radix aqueous extract 5:4:1 (g/g) mixture on the primary cultured rat articular chondrocytes. First, any cytotoxic effect of mixture was observed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium Bromide) assay. Next, cyto-protective effect of test substances was evaluated by using the recombinant human interleukin $(rhIL)-1{\alpha}$ induced chondrocytes. In addition, anti-inflammatory effects were also observed on the lipopolysaccaride (LPS) treated chondrocytes through prostaglandin $E_2(PGE_2)$ productions and 5-lipoxygenase (LPO) activities, and inhibitory effects on matrix metalloproteinase (MMP)-2 and MMP-9 activities were observed on $rhIL-1{\alpha}$ treated chondrocytes with their extracellular matrix (ECM) related mRNA expressions. No obvious cytotoxic effects of mixture were demonstrated. Inflammatory damages of chondrocytes and related ECM degradations induced by treatment of LPS or $rhIL-1{\alpha}$ were significantly and concentration-dependently inhibited by pretreatment of mixture from a concentration level of 0.001 mg/ml to 1 mg/ml. In addition, mixture showed $IC_{50}$ for $rhIL-1{\alpha}-induced$ MMP-2 and MMP-9 activities as 44.01 and $162.47{\mu}g/ml$, and also showed $EC_{50}$ for $rhIL-1{\alpha}-induced$ inhibition of collagen type II, SOX9 and aggrecan mRNA expression as 8.61, 10.79 and $4.47{\mu}g/ml$, respectively. It is observed that mixture showed concentration-dependent anti-inflammatory and cytoprotective ECM preserved effects on the primary cultured rat articular chondrocytes without cytotoxicity.

• Asian-Australasian Journal of Animal Sciences
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• 제32권2호
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• pp.257-264
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• 2019

Objective: Dairy cattle nutrient requirement systems acknowledge amino acid (AAs) requirements in aggregate as metabolizable protein (MP) and assume fixed efficiencies of MP used for milk protein. Regulation of mammary protein synthesis may be associated with AA input and milk protein output. The aim of this study was to evaluate the effect of nanoemulsified methionine and cysteine on the in-vitro expression of milk protein (casein) in bovine mammary epithelial cells (MAC-T cells). Methods: Methionine and cysteine were nonionized using Lipoid S 75 by high-speed homogenizer. The nanoemulsified AA particle size and polydispersity index were determined by dynamic light scattering correlation spectroscopy using a high-performance particle sizer instrument. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to determine the cytotoxicity effect of AAs with and without nanoionization at various concentrations (100 to $500{\mu}g/mL$) in mammary epithelial cells. MAC-T cells were subjected to 100% of free AA and nanoemulsified AA concentration in Dulbecco's modified Eagle medium/nutrient mixture F-12 (DMEM/F12) for the analysis of milk protein (casein) expression by the quantitative reverse transcription polymerase chain reaction method. Results: The AA-treated cells showed that cell viability tended to decrease (80%) in proportion to the concentration before nanogenesis, but cell viability increased as much as 90% after nanogenesis. The analysis of the expression of genetic markers related to milk protein indicated that; ${\alpha}_{s2}$-casein increased 2-fold, ${\kappa}$-casein increased 5-fold, and the amount of unchanged ${\beta}$-casein expression was nearly doubled in the nanoemulsified methionine-treated group when compared with the free-nanoemulsified methionine-supplemented group. On the contrary, the non-emulsified cysteine-administered group showed higher expression of genetic markers related to milk protein ${\alpha}_{s2}$-casein, ${\kappa}$-casein, and ${\beta}$-casein, but all the genetic markers related to milk protein decreased significantly after nanoemulsification. Conclusion: Detailed knowledge of factors, such nanogenesis of methionine, associated with increasing cysteine and decreasing production of genetic markers related to milk protein (casein) will help guide future recommendations to producers for maximizing milk yield with a high level of milk protein casein.

• 한방안이비인후피부과학회지
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• 제32권3호
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• pp.37-47
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• 2019

Objectives : The purpose of this study is to investigate the anti-oxidative and the anti-inflammatory effects of Danpitang(DPT) extract in RAW 264.7 macrophages. Methods : The macrophage cell line RAW 264.7 cells were used and MTT assay was performed to measure the cell viabilities at the various concentrations of DPT($50-400{\mu}g/m{\ell}$). Nitric oxide(NO) was measured in LPS-induced RAW 264.7 cells. Expressions of iNOS, COX-2, $TNF-{\alpha}$, $IL-1{\alpha}$, $IL-1{\beta}$ and IL-6 were also performed by real-time PCR. Protein expression of iNOS and COX-2 was confirmed by western blot. The anti-oxidant activities of DPT was measured by DPPH radical scavenging activity. Results : 1. There was no cytotoxicity in RAW 264.7 cells treated with DPT compared to the control. 2. DPT treated group significantly inhibited NO production compared to the LPS treated group. 3. DPT treated group significantly decreased mRNA expressions of iNOS, COX-2, $TNF-{\alpha}$, $IL-1{\alpha}$, $IL-1{\beta}$ and IL-6 compared to the LPS treated group. 4. To evaluate the safety of the products for the human body, Adverse events, SCORAD Index Assessment were conducted; There were no severe adverse events during this study. And SCORAD Index showed a statistically significant decrease in treatment group in baseline, 2 weeks and 4 weeks. Therefore, it is suggested that products, if used for certain period, should be safe for the human body. 5. DPT was found to have high DPPH free radical scavenging ability. Conclusions : According to the above results, DPT can be used as a therapy in various anti-inflammatory skin diseases.

• 대한약침학회지
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• 제22권3호
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• pp.176-183
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• 2019

Objective: In the current work, we investigated the cytotoxic and apoptotic effects of Thymoquinone (TQ), an active compound of Nigella sativa (N. sativa) and Cis-platinum, on normal renal epithelial (GP-293) and human renal adenocarcinoma cell lines (ACHN). Methods: GP-293 and ACHN cell lines were cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% Fetal bovine serum (FBS) and 1% penicillin plus streptomycin antibiotic. The MTT assay was used for cellular viability assessment. Viability of cells was observed using inverted light microscope 24, 48 and 72 h after exposure of the cells to various concentrations of TQ (1, 2.5, 5, 10, 50 and $100{\mu}g/ml$) and Cis-platinum (0.5, 1, 1.5, 2, 3, 6 and $12.5{\mu}g/ml$). Moreover, apoptosis was analyzed with a flow-cytometry method. The untreated cells were considered as control group. Results: Morphological changes such as reduced cell number and increased intercellular distance and reduced cell viability in ACHN and GP-293cell lines were observed in both TQ and Cis- platinum groups; however, Cis-platinum had greater effect on ACHN cell line than GP-293 cell line. In addition, GP-293 cell line was more sensitive to TQ compared to ACHN cell line. Furthermore, TQ and Cis-platinum had apoptotic effects on both ACHN and GP-293 cell lines. Conclusion: Our findings demonstrated that TQ and Cis-platinum had cytotoxic and apoptotic effects on both cell lines, However, GP-293 cell line was more sensitive to TQ. Additionally, Cis-platinum was more effective on ACHN cell line than on GP-293 cell line.

• 생약학회지
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• 제50권1호
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• pp.11-17
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• 2019

Cassia tora L. have been used as a folk medicine in Korea. This study investigated anti-inflammatory effect of aurantio-obtusin isolated from C. tora. We isolated aurantio-obtusin from 50% ethanol extracts of C. tora L. We investigated the anti-inflammatory effects of aurantio-obtusin on the lipopolysaccharide (LPS)-stimulated inflammatory response in murine macrophage cell line (Raw 264.7). To investigate the cytotoxicity of aurantio-obtusin on RAW 264.7 cells, MTS assay was performed. RAW 264.7 cells were treated with aurantio-obtusin at different concentrations (12.5, 25, 50, $100{\mu}M$) for 30 h. The result showed that aurantio-obtusin had no cytotoxic effect in a concentration range of $12.5-100{\mu}M$. To determine the effect of aurantio-obtusin on LPS-induced NO production, the NO concentration measurement was performed. RAW 264.7 cells were treated with aurantio-obtusin at 12.5, 25, 50 and $100{\mu}M$ for 24 h, and the results showed that the NO production of aurantio-obtusin-treated cells compared to LPS alone treated group was significantly decreased in a dose-dependent manner. Pretreatment of aurantio-obtusin inhibited LPS-induced NO production in a dose-dependent manner. To find out inhibitory mechanisms of aurantio-obtusin on inflammatory mediators, we examined the $PGE_2$ pathways. As a result, $PGE_2$ were decreased in a dose-dependent manner by aurantio-obtusin. The release of interleukin-$1{\beta}$ (IL-$1{\beta}$) and IL-6 were also reduced. Moreover, aurantio-obtusin suppressed LPL-induced $I{\kappa}B-{\alpha}$ degradation. These results suggest that the down regulation of NO, $PGE_2$, IL-$1{\beta}$ and IL-6 expression by aurantio-obtusin are achieved by the downregulation of NF-${\kappa}B$ activity.

• Journal of Nutrition and Health
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• 제54권5호
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• pp.448-458
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• 2021

본 연구는 뽕나무 열매 착즙 분말 (MJ)이 지방세포 분화과정에서 염증에 관여하는 유전자 발현 및 miR-132/143 조절에 미치는 효과에 중점을 두고 조사하였다. MJ는 3T3-L1 지방세포 분화 동안 지질축적이 억제되었고 PPAR-γ, CEBP-α 및 aP2와 같은 지방형성 유전자 발현을 억제하였다. 또한 염증 조절 매개체인 TNF-α, IL-6, MCP-1 및 iNOS 유전자 발현이 MJ 처리에 의해 감소되었다. MJ의 지질 축적 억제 효과는 염증과 지방분화에 관여하는 miR-132/143의 발현 감소와 관련이 있음을 확인하였다. 따라서 MJ는 염증을 동반하는 비만 예방 및 치료에 도움이 될 수 있는 식품 소재로서 가능성이 있음을 시사하고 있다. 그러나 MJ의 주요 성분인 안토시아닌의 생체이용율은 1-2% 미만으로 추정되며 예상되는 표적 조직이나 혈류에서 미량 검출되는 것으로 알려져 있다 [33]. 이를 해결하기 위해서는 생체 내 동물실험을 통한 다각적인 분석이 향후 진행되어야 할 것으로 판단된다.

• 생명과학회지
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• 제31권2호
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• pp.175-182
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• 2021

기능성 화장품 소재로서 활용할 수 있는 효모의 분리를 위하여 순창군 베리류 및 과수원 토양에서 분리주 80종을 1차로 선별하였다. 80종의 분리주를 대상으로 항산화 활성 및 tyrosinase 저해 활성을 측정한 결과 DPPH 라디칼 소거능은 51.41%, SOD 활성은 62.23%, tyrosinase 저해 활성 64.75%로 가장 우수한 FT4-4를 최종적으로 선별하였다. 18S rRNA 염기서열 분석을 통해 Saccharomyces cerevisiae FT4-4로 명명하였으며, API ZYM을 이용하여 세포 외 효소 활성을 추가로 측정하였다. 발효 시간에 따른 균체 성장 및 tyrosinase 저해 활성의 변화를 측정한 결과 배양 후 16시간에 최대 균체량인 3.16 g/l와 67.68%의 tyrosinase 저해활성을 나타내었다. 또한, S. cerevisiae FT4-4의 화장품 소재로 활용하기 위한 세포 독성과 melanoma B16F10 세포 멜라닌 억제능을 확인한 결과, 세포독성은 50 mg/ml 이하의 농도에서 100% 이상의 세포 생존율을 보였으며, 시료 10 mg/ml에서의 멜라닌 생합성 저해능은 72.02%로 측정되었다. 향후 FT4-4의 화장품 소재로 활용하기 위해서는 생산 수율을 증가하기 위한 생산조건 확립 이외에도 안전성을 향상시키기 위한 추가적인 독성연구 등 다양한 연구가 수반되어야 하나, 본 연구에서의 항산화 및 미백 효능만으로도 충분히 활용할 가치가 있는 소재로 사료된다.

• Journal of Applied Biological Chemistry
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• 제63권4호
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• pp.327-334
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• 2020

산화적 스트레스는 신경퇴행성 질환 발병의 원인으로 알려져 있다. 본 연구는 대표적인 감초 종류인 Glycyrrhiza glabra, G. uralensis와 신품종 감초인 신원감(SW)의 in vitro free radical 소거능을 통한 항산화 활성과 H2O2 유도 산화적 스트레스에 대한 C6 glial cell 보호 효능을 확인하고자 하였다. In vitro assay에서 G. uralensis, G. glabra, SW 추출물은 농도유의적으로 2,2-diphenyl-1-picrylhydrazyl, ·OH, O2- radical 소거능이 증가하여 in vitro 항산화 활성을 확인하였다. 또한, SW 추출물은 G. uralensis, G. glabra 추출물에 비해 총 페놀 및 플라보노이드 함량이 가장 우수하였다. H2O2로 산화적 스트레스를 유도한 C6 glial cell에 3가지 감초 추출물을 각각 처리 시, 농도의존적으로 세포 생존율이 증가와 reactive oxygen species 소거능이 증가하여 3가지 감초 추출물의 산화적 손상에 대한 신경교세포 보호 효과를 확인하였다. 특히, SW 추출물은 G. uralensis, G. glabra 추출물에 비해 우수하게 C6 glial cell 보호 효과를 나타내었다. 또한, 3가지 감초 추출물의 신경교세포 보호 메커니즘을 확인하기 위해, 염증 관련 단백질 발현을 측정하였다. 3가지 감초 추출물은 H2O2만을 처리한 control군에 비해 inducible nitric oxide synthase 및 cyclooxygenase-2 발현 감소를 통해 염증반응 조절을 통한 신경교세포 보호 작용기전을 확인하였다. 본 연구는 G. uralensis, G. glabra, SW 등 3가지 감초 추출물이 산화적 손상이 유도된 신경교세포 보호에 유용한 소재로써의 가능성이 있는 것으로 사료된다.

• Journal of Nutrition and Health
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• 제55권1호
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• pp.59-69
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• 2022

본 연구실의 선행 연구에서 항산화 활성이 가장 높았던 조팝나무 H₂O-EtOH (40:60) 분획물(SP60)의 항염 효능을 확인하여 기능성 소재로의 개발 가능성을 알아보고자 하였다. 항염활성 측정을 위해 RAW 264.7 세포에서 먼저 WST assay를 실시하여 SP60이 125 ㎍/mL 농도까지 세포 생존율에 영향을 미치지 않는 것을 확인하였다. Raw264.7 세포에 LPS와 SP60을 동시에 처리하여 NO, TNF-α, IL-6, IL-1β 생성과 iNOS, COX-2, NF-κB 발현을 측정하였다. SP60은 NO, TNF-α, IL-6, IL-1β 생성량을 모두 유의하게 억제시켰으며 western blot으로 iNOS, COX-2의 발현과 NF-κB의 활성을 측정한 결과 SP60을 처리한 군에서 농도 의존적으로 그 발현량과 활성을 억제하였다. 따라서 조팝나무 분획물은 항염 효능을 가진 것으로 판단되며 항염 관련 기능성 식품 및 소재로의 개발 가능성을 확인하였다.