• 한국식품위생안전성학회지
• /
• 제32권3호
• /
• pp.243-248
• /
• 2017

현재 식품공전에 식품원료로 사용이 가능한 것으로 등록된 국내 산림지역 자생 식물을 식품산업에 활용하고자 국내 자생식물 45종을 선별하였고, 본 연구팀의 선행연구에서 45종의 항산화 활성을 평가한 결과, 다른 종들과 비교하여 우수한 항산화 활성을 보인 밤나무 잎을 본 연구에서 사용하였다. 본 연구에서는 hydrogen peroxide로 산화적 스트레스를 유도한 상태에서 밤나무 잎 추출물의 세포 보호효과, 세포내 항산화 효과 및 항노화 효과를 관찰하였다. 세포 보호효과를 측정한 결과 밤나무 잎 추출물의 농도 유의적으로 세포를 보호하는 효과를 확인하였고, 특히 $100{\mu}g/mL$에서는 양성대조군으로 사용한 항산화 물질인 ascorbic acid 수준까지 세포를 보호하는 효과를 나타내었다. $H_2$-DCFDA 염색을 통한 세포내 항산화 효과를 확인한 결과, 형광현미경과 형광흡광도 측정에서 모두 밤나무 잎 추출물이 농도 유의적으로 세포내 ROS를 저감을 확인하였고, 세포 보호효과와 마찬가지로 농도 $100{\mu}g/mL$에서는 ascorbic acid와 비슷한 수준을 보였다. SA-${\beta}$-galactosidase 염색을 통하여 관찰한 세포노화 억제 효과는 ROS 생성 억제 효과와 유사한 경향으로 밤나무 잎 추출물의 농도 유의적으로 세포노화 억제를 관찰되었다. 이상의 결과를 종합하여 볼 때 밤나무 잎 추출물은 산화적 스트레스로 인한 세포노화를 억제하는 효능을 보였으며, 이는 천연물에서 유래한 항산화 및 항노화 활성 첨가제로 활용도가 매우 넓을 것으로 판단된다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• 제34권2호
• /
• pp.220-229
• /
• 2008

Purpose: The present study was aimed to examine the effect of acellular dermal matrix ($AlloDerm^{(R)}$) grafted to the experimental tissue defect on tissue regeneration. Materials and Methods: Male albino rabbits were used. Soft tissue defects were prepared in the external abdominal oblique muscle. The animals were then divided into 3 groups by the graft material used: no graft, autogenous dermis graft, and $AlloDerm^{(R)}$ graft. The healing sites were histologically examined at weeks 4 and 8 after the graft. In another series, critical sized defects with 8-mm diameter were prepared in the right and left iliac bones. The animals were then divided into 5 groups: no graft, grafted with autogenous iliac bone, $AlloDerm^{(R)}$ graft, $AlloDerm^{(R)}$ graft impregnated with rhBMP-2, and $AlloDerm^{(R)}$ graft with rhTGF-${\beta}1$. The healing sites of bone defect were investigated with radiologic densitometry and histological evaluation at weeks 4 and 8 after the graft. Results: In the soft tissue defect, normal healing was seen in the group of no graft. Inflammatory cells and foreign body reactions were observed in the group of autogenous dermis graft, and the migration of fibroblasts and the formation of vessels into the collagen fibers were observed in the group of $AlloDerm^{(R)}$ graft. In the bone defect, the site of bone defect was healed by fibrous tissues in the group of no graft. The marked radiopacity and good regeneration were seen in the group of autogenous bone graft. There remained the traces of $AlloDerm^{(R)}$ with no satisfactory results in the group of $AlloDerm^{(R)}$ graft. In the groups of the $AlloDerm^{(R)}$ graft with rhBMP-2 or rhTGF-${\beta}1$, there were numerous osteoblasts in the boundary of the adjacent bone which was closely approximated to the $AlloDerm^{(R)}$ with regeneration features. However, the fibrous capsule also remained as in the group of $AlloDerm^{(R)}$ graft, which separated the $AlloDerm^{(R)}$ and the adjacent bone. Conclusions: These results suggest that $AlloDerm^{(R)}$ can be useful to substitute the autogenous dermis in the soft tissue defect. However, it may not be useful as a bone graft material or a carrier, since the bone defect was not completely healed by the bony tissue, regardless of the presence of osteogenic factors like rhBMP-2 or rhTGF-${\beta}1$.

• Biomolecules & Therapeutics
• /
• 제28권4호
• /
• pp.354-360
• /
• 2020

The hair cycle (anagen, catagen, and telogen) is regulated by the interaction between mesenchymal cells and epithelial cells in the hair follicles. The proliferation of dermal papilla cells (DPCs), mesenchymal-derived fibroblasts, has emerged as a target for the regulation of the hair cycle. Here, we show that vanillic acid, a phenolic acid from wheat bran, promotes the proliferation of DPCs via a PI3K/Akt/Wnt/β-catenin dependent mechanism. Vanillic acid promoted the proliferation of DPCs, accompanied by increased levels of cell-cycle proteins cyclin D1, CDK6, and Cdc2 p34. Vanillic acid also increased the levels of phospho(ser473)-Akt, phospho(ser780)-pRB, and phospho(thr37/46)-4EBP1 in a time-dependent manner. Wortmannin, an inhibitor of the PI3K/Akt pathway, attenuated the vanillic acid-mediated proliferation of DPCs. Vanillic acid-induced progression of the cell-cycle was also suppressed by wortmannin. Moreover, vanillic acid increased the levels of Wnt/β-catenin proteins, such as phospho(ser9)-glycogen synthase kinase-3β, phospho(ser552)-β-catenin, and phospho(ser675)-β-catenin. We found that vanillic acid increased the levels of cyclin D1 and Cox-2, which are target genes of β-catenin, and these changes were inhibited by wortmannin. To investigate whether vanillic acid affects the downregulation of β-catenin by dihydrotestosterone (DHT), implicated in the development of androgenetic alopecia, DPCs were stimulated with DHT in the presence and absence of vanillic acid for 24 h. Western blotting and confocal microscopy analyses showed that the decreased level of β-catenin after the incubation with DHT was reversed by vanillic acid. These results suggest that vanillic acid could stimulate anagen and alleviate hair loss by activating the PI3K/Akt and Wnt/β-catenin pathways in DPCs.

• 대한화장품학회지
• /
• 제42권2호
• /
• pp.163-171
• /
• 2016

스테로이드 호르몬인 glucocorticoid (GC)는 glucocorticoid receptor (GR)와 결합하여 염증 유전자의 발현을 억제함으로써 강력한 항염 효과를 준다. 따라서 GR을 활성화하는 GC 제제들이 개발되어 피부염의 치료제로 사용되어 왔다. 그러나 이러한 GC 제제들은 피부 장벽 기능 저하, 진피층 두께 감소 등의 부작용을 유발하여 피부를 손상시키고 피부 노화를 유발한다. 특히 GC 성분은 11 beta-hydroxysteroid dehydrogenase type 1 ($11{\beta}$-HSD1)에 의해 활성화되어 GR의 활성을 높이는 것으로 보고되어 있다. 이에 본 연구에서는 스테로이드 제제인 dexamethasone에 의해 증가된 $11{\beta}$-HSD1의 발현을 효과적으로 억제할 수 있는 천연 소재를 발굴하고자 하여, 초두구 추출물에서 유래한 alpinetin에서 유의한 효과가 있음을 확인하였다. Alpinetin은 진피 섬유아세포에서 dexamethasone에 의해 발현이 증가한 $11{\beta}$-HSD1를 억제함과 동시에 GR의 활성 및 cortisol의 생성을 감소시켰다. 또한 사람 섬유아세포 및 3D skin model을 이용한 평가에서, alpinetin은 dexamethasone에 의한 콜라겐 감소와 진피층 두께 감소를 효과적으로 회복시켰다. 따라서, 본 연구 결과로부터 alpinetin은 $11{\beta}$-HSD1의 발현 증가에 의한 피부 스트레스 및 피부 노화를 효과적으로 예방할 수 있을 것으로 사료되었다.

• Journal of Periodontal and Implant Science
• /
• 제32권1호
• /
• pp.129-142
• /
• 2002

Epithelial-mesenchymal interaction plays a important role in cell growth and differentiation. This interaction is already well known to have an importance during the organ development as well as cell growth and differentiation. However, in vitro experimental model is not well developed to reproduce in vivo cellular microenvironment which provide a epithelial-mesenchymal interaction. Because conventional monolayer culture lacks epithelial-mensenchymal interaction, cultivated cells have an morphologic, biochemical, and functional characteristics differ from in vivo tissue. Moreover, it's condition is not able to induce cellular differention due to submerged culture condition. Therefore, the aims of this study were to develop and evaualte the in vitro experimental model that maintains epithelial-mesenchymal interaction by organotypic raft culture, and to characterize biologic properties of three-dimensionally reconstituted oral keratinocytes by histological and immunohistochemical analysis. The results were as follow; 1. Gingival keratinocytes reconstituted by three-dimensional organotypic culture revealed similar morphologic characteristics to biopsied patient specimen showing stratification, hyperkeratinosis, matutation of epithelial architecture. 2. Connective tissue structure was matured, and there is no difference during stratification period of epithelial 3-dimensional culture. 3. The longer of air-exposure culture on three-dimensionally reconstituted cells, the more epithelial maturation, increased epithelial thickness and surface keratinization 4. In reconstitued mucosa, the whole epidermis was positively stained by anti-involucrin antibody, and there is no difference according to air-exposured culture period. 5. The Hsp was expressed in the epithelial layer of three-dimensionally cultured cells, especially basal layer of epidermis. The change of Hsp expression was not significant by culture stratification. 6. Connexin 43, marker of cell-cell communication was revealed mild immunodeposition in reconstitued epithelium, and there is no significant expression change during stratification. These results suggest that three-dimensional oragnotypic co-culture of normal gingival keratinocytes with dermal equivalent consisting type I collagen and gingival fibroblasts results in similar morphologic and immunohistochemical characteristics to in vivo patient specimens. And this culture system seems to provide adequate micro-environment for in vitro tissue reconstitution. Therefore, further study will be focused to study of in vitro gingivitis model, development of novel perioodntal disease therapeutics and epithelial-mensenchymal interaction.

• 대한화장품학회지
• /
• 제31권4호
• /
• pp.323-327
• /
• 2005

노화에 미치는 자외선의 영향에 대해 많은 연구가 진행되어 왔지만, 천연물에 대한 연구는 별로 알려진 것이 없다. MMPs는 광노화 과정에 매우 중요한 역할을 하는 것으로 알려져 있다. 본 연구에서는 MMP-1의 발현과 활성 및 항산화 효과에 미치는 $1,2,4,6-tetra-O-galloyl-{\beta}-{_D}-glucopyranose$와 3,4,5-trihydroxybenzoic acid의 효과를 측정하였다. 이들 화합물은 박과의 백렴으로부터 분리하였으며, 자유 라디칼과 활성산소 소거활성이 매우 높은 것으로 나타났다. 이들 화합물의 DPPH 라디칼과 활성산소를 50% 소거하는 활성$(SC_{50})$은 각각 $3.9{\mu}M,\;13.3{\mu}M,\;4.3{\mu}M,\;4.0{\mu}M$로 나타났다. 또한, 이들 화합물은 단백질 수준에서의 MMP-1 발현 및 활성을 처리농도 의존적으로 저해하였지만, 유전자 수준에서는 저해활성이 없는 것으로 나타났다 따라서, 이들 화합물은 단백질 수준에서의 우수한 MMP-1 발현 저해능과 높은 항산화 활성을 가지는 것을 알 수 있다. 결론적으로 이들 화합물은 새로운 항노화 소재로 적용될 수 있을 것으로 기대된다

• 대한화장품학회지
• /
• 제48권2호
• /
• pp.157-167
• /
• 2022

본 연구에서는 7 개의 아미노산으로 이루어진 헵타펩타이드의 섬유아세포 활성 증가 및 광노화 조건에서의 세포 손상 억제 효과를 확인하였다. 실험 결과 헵타펩타이드 처리 시 섬유아세포 증식 및 세포외기질(extracellular matrix, ECM) 구성 인자의 발현이 증가되었다. 그리고 자외선 A (ultraviolet A, UVA) 조사에 의해 유도된 광노화조건에서 감소된 세포 생존율이 헵타펩타이드에 의해 증가되었고, UVA 조사에 의해 유도된 세포 사멸, 기질금속단백질분해효소-1(matrix metalloproteinases-1, MMP-1) 발현 및 세포 내 활성산소종(reactive oxygen species, ROS) 수준이 헵타펩타이드에 의해 감소되었다. UVA 조사 시 나타나는 transforming growth factor-β (TGF-β)/smad 기전 억제와 그에 따른 ECM 구성 인자 발현 감소 또한 헵타펩타이드에 의해 회복되었다. 또 다른 광노화 유도 조건으로 heat shock을 주었고 헵타펩타이드를 전 처리 하였을 때 heat shock에 의한 mitogen-activated protein kinase (MAPK) 인산화 및 MMP-1 발현이 억제됨을 확인할 수 있었다. 이 결과를 종합해 볼 때, 본 연구의 헵타펩타이드는 섬유아세포의 활성을 촉진하며, 광노화 유도 모델로 사용된 UVA 조사 및 heat shock 조건에서도 세포 내 ROS 억제 효과를 보여 세포 손상에 대한 회복 및 보호 효과를 나타내는 것으로 보인다. 이러한 진피 보호 효과를 갖는 헵타펩타이드는 향 후 신규 화장품 소재로 응용될 수 있을 것으로 기대된다.

• Toxicological Research
• /
• 제34권1호
• /
• pp.31-39
• /
• 2018

The objective of this study was to evaluate the antioxidant and anti-aging effects of marigold methanol extract (MGME) in human dermal fibroblasts. Total polyphenolic and flavonoid contents in MGME were 74.8 mg TAE (tannic acid equivalent)/g and 85.6 mg RE (rutin equivalent)/g, respectively. MGME ($500{\mu}g/mL$) increased 1,1-diphenyl-2-picryl hydrazyl (DPPH) and 2,2'-azino-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical-scavenging, and superoxide dismutase (SOD)-like antioxidant activities by 36.5, 54.7, and 14.8%, respectively, compared with the control. At $1,000{\mu}g/mL$, these activities increased by 63.7, 70.6, and 20.6%, respectively. MGME ($100{\mu}g/mL$) significantly increased the synthesis of type 1 procollagen by 83.7% compared with control treatment. It also significantly decreased Matrix Metalloproteinase-2 (MMP-2) activity and MMP-1 mRNA expression by 36.5% and 69.5%, respectively; however, it significantly increased laminin-5 mRNA expression by 181.2%. These findings suggest that MGME could protect human skin against photo-aging by attenuating oxidative damage, suppressing MMP expression and/or activity as well as by stimulating collagen synthesis.

• BMB Reports
• /
• 제48권8호
• /
• pp.467-472
• /
• 2015

Heat shock increases skin temperature during sun exposure and some evidence indicates that it may be involved in skin aging. The antioxidant response mediated by the transcription factor NF-E2-related factor 2 (Nrf2) is a critically important cellular defense mechanism that serves to limit skin aging. We investigated the effects of heat shock on collagenase expression when the antioxidant defense system was downregulated by knockdown of Nrf2. GSH and collagenases were analyzed, and the expression of inducible Nrf2, HO-1, and NQO1 was measured. HS68 cells were transfected with small interfering RNA against Nrf2. Heat shock induced the downregulation of Nrf2 in both the cytosol and nucleus and reduced the expression of HO-1, GSH, and NQO1. In addition, heat-exposed Nrf2-knockdown cells showed significantly increased levels of collagenase protein and decreased levels of procollagen. Our data suggest that Nrf2 plays an important role in protection against heat shock-induced collagen breakdown in skin. [BMB Reports 2015; 48(8): 467-472]

• 생약학회지
• /
• 제44권1호
• /
• pp.47-52
• /
• 2013

We invesitagat the effects of antioxidant and suppression of MMP-1 in UV-irradiated human dermal fibroblasts (HDF) by schizandrin isolated from Schisandra chinensis. Ultrasonic technique was used to extract schizandrin from S. chinensis. The higher contents of schizandrin (6.34 mg/ml) in 70% EtOH extract obtained by ultrasonic treatment at 20 MHz for 2 h during 10 days. Schizandrin had antioxidant activities. Schizandrin (1-10 ${\mu}g/ml$) induced a significant dose-dependent decrease in the expression fo MMP-1 protein. These results suggest that schizandrin is a potential candidate for the prevention and treatment of skin aging.