• 제목/요약/키워드: Human Fibroblast

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유산균 발효에 의한 인삼열매 추출물의 항산화 및 항노화 효과 (Antioxidant and Antiaging Effect of Ginseng Berry Extract Fermented by Lactic Acid Bacteria)

  • 전지민;최성규;김윤정;장수진;천종우;이현상
    • 대한화장품학회지
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    • 제37권1호
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    • pp.75-81
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    • 2011
  • 본 연구에서는 유산균 발효에 의한 인삼열매 추출물의 여러 가지 생리활성을 평가하였다. 고성능액체크로마토그래피를 수행하여 인삼열매 추출물과 유산균 발효 인삼열매 추출물의 ginsenoside Re, Rc 및 Rb1의 함량분석 결과 유산균 발효에 의해 ginsenoside Re, Rc 및 Rb1의 함량이 모두 증가하였다. 인삼열매 추출물과 유산균 발효 인삼열매 추출물의 DPPH free radical 소거활성과 SOD 유사활성 측정결과 1.00 %의 농도에서 유산균 발효 인삼열매 추출물은 각각 86.34, 76.82 %의 DPPH free radical 소거활성과 SOD 유사활성을 나타내었고, 인삼열매 추출물은 각각 49.78, 40.80 %의 DPPH free radical 소거활성과 SOD 유사활성을 나타내어 유산균 발효에 의해 항산화 활성이 증가하는 것을 확인하였다. 또한 유산균 발효 인삼열매 추출물 0.50 %의 농도로 처리한 피부 섬유아세포에서 procollagen type I (COL1A1)은 828.13 % 증가하였으며, matrix metalloproteinase (MMP)-1은 87.88 %, tumor necrosis factor (TNF)-${\alpha}$는 99.92 %감소하는 효과를 보여주었다. 이처럼 항산화, 주름, 항염에 효능을 보이는 유산균 발효에 의한 인삼열매 추출물은 기능성 화장품 소재로 개발될 수 있는 가능성이 있다.

백수오(白首烏)의 항산화 효능 및 모유두 세포의 5α-reductase mRNA 발현 억제 효능 (Investigation of Antioxidant Activity of Cynanchi Wilfordii Radix and Inhibitory Effect of 5α-reductase mRNA in Human Dermal Papilla Cells)

  • 조남준;최영호;전해리;이웅희;김기광;한효상
    • 동의생리병리학회지
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    • 제31권6호
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    • pp.374-379
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    • 2017
  • Hair loss affects interpersonal relationships and causes psychological stress. In this study, we investigated the antioxidant activity of Cynanchi Wilfordii Radix (CWR) and its effects on dermal papilla (DP) cells. Antioxidant efficacy was examined by ABTS assay. To confirm the effect on cell activity, MTS assay was performed and cell count was directly measured by hemocytometer. The mRNA expression of genes involved in hair formation and hair loss formation was confirmed by quantitative RT-PCR. CWR has a strong antioxidant activity. Cell viability of DP cells was increased to 118.5% by treatment of 0.5 mg/ml CWR for 24 hours, but the effect on the cell number was insignificant. These results suggest that CWR increases mitochondrial activity without promoting cell proliferation. Treatment of DP cells with 0.5 mg/ml CWR resulted in 48.5% reduction of mRNA expression of type 2 $5{\alpha}$-reductase, a major cause of male hair loss. In addition, mRNA expression of bone morphogenetic pretein (BMP), fibroblast growth factor (FGF)7, and FGF10, which are closely related to hair growth, was also decreased. Reactive oxygen species (ROS) acts as a cause of hair loss. The excellent antioxidant efficacy of CWR is thought to be able to effectively remove ROS. The dihydrotestosterone produced by type 2 $5{\alpha}$-reductase in DP cells is a potent inducer of male pattern hair loss. The inhibitory effect of type 2 $5{\alpha}$-reductase mRNA on DP cells induced by CWR may induce a positive therapeutic effect of male pattern hair loss.

교정용 호선의 세포독성에 관한 실험적 연구 (An experimental study on the cytotoxicity of orthodontic wires)

  • 임용규;양원식
    • 대한치과교정학회지
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    • 제26권5호
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    • pp.591-599
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    • 1996
  • 본 연구는 널리 이용되고 있는 두 종류의 교정용 호선에 다양한 처리를 가한 후 호선의 세포독성을 비교, 평가하고자 시행하였다. 018x025 inch 굵기의 stainless steel 호선과 Co-Cr 호선을 실험 재료로 선택하여 stainless steel 호선을 A 호선, Co-Cr호선을 B 호선이라 칭하였으며 각각의 호선을 다시 가해진 처리에 따라 4군으로 나누었다. A-1군과 B-1군은 제작된 상태 그대로의 호선을 이용하였으며 A-2, B-2군은 기기를 이용하여 $850^{\circ}\;F$에서 4분간 열처리하였다. A-3, B-3군은 같은 방법으로 열처리한 후 표면의 불순물을 제거하기 위해 전해연마를 시행하였고 A-4, B-4군은 소량의 은납(Ag-solder)을 납착(soldering) 하였다. 사람의 치은 섬유아세포를 배양하고 agar overlay법을 이용하여 각군의 호선의 세포독성을 검사하였으며 세포독성을 반응지수(탈색지수/용해지수)로 평가하여 다음의 결론을 얻었다. 1. stainless steel 호선과 Co-Cr 호선 모두 제작된 그대로의 상태에서는 세포독성을 나타내지 않았다. 2. 두 호선에 대한 열처리나 전해연마는 호선의 세포독성에 영향을 미치지 못하였다. 3. 은납이 납착된 stainless steel호선은 은납이 납착된 Co-Cr호선에 비해 더 넓은 범위의 탈색을 나타냈으나 탈색지수와 세포독성(반응지수)에서는 차이를 보이지 않았다. 4. 은납이 납착된 호선은 두 호선 모두에서 중증도의 세포독성을 나타냈다.

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발효 공정을 통한 아로니아 추출물의 항염증 효능 증진 (Enhancement of Anti-inflammation Effect by Fermentation Process in Aronia melanocarpa (Michx.) Elliott Extract)

  • 김남영;이영덕;조석철;신윤철;이현용
    • 한국약용작물학회지
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    • 제22권6호
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    • pp.475-482
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    • 2014
  • This study was indicated to enhance the anti-inflammation activities by the fermentation of the fruits of Aronia melanocarpa (Michx.) Elliott. The extracts by 70% ethanol (EE) showed better biological activities than those by hot water (WE) from campared result of the effect of extraction solvents. Then, the extract from 70% ethanol extraction was further fermented by lactic acid, denoted as FEE. For antioxidant activities, the FEE had showed the highest value as 0.832 of reducing powder, in comparison with those of EE and WE. Cytotoxicity of the water extraction (WE) was measured for 12.06% in addition of $1.0mg/m{\ell}$ of FEE. For anti-inflammation activities, NO production from the macrophage, RAW 264.7 was observed as $7.24{\mu}M$ and $8.52{\mu}M$ from FEE and EE, respectively. Prostaglandin $E_2$ ($PGE_2$) production from human fibroblast cell, CCD-986sk, was also estimated for $152pg/m{\ell}$ in addition of $1.0mg/m{\ell}$ of the FEE. The lowest production of both IL-6 and TNF-${\alpha}$ were $3.5pg/m{\ell}$ and $865.5pg/m{\ell}$, respectively in addition of $1.0mg/m{\ell}$ of the FEE, whereas $74.5pg/m{\ell}$ and $982.4pg/m{\ell}$ in treated with same concenrations of the EE. It was also found that the FEE was higher amounts than other extracts through HPLC analysis of the anthocyanins. These results strongly indicate that fermentation process of the lactic acid could enhance anti-inflammation activities of extracts by increasing the amounts of the anthocyanins, especially cyanidin-galactoside. Our results suggest that the application of the fermentation process for other medicinal herbs can be improved their biological activities.

생쥐의 간과 HepG2 세포에 있어서 내인성 small heterodimer partner (SHP)의 단백질 수준에 미치는 cholic acid/CDCA 및 FGF-19의 효과 (Effects of Cholic Acid/CDCA and FGF-19 on the Protein Levels of the Endogenous Small Heterodimer Partner (SHP) in the Mouse Liver and HepG2 Cells)

  • 민계식
    • 생명과학회지
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    • 제19권12호
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    • pp.1731-1736
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    • 2009
  • 최근의 연구에서 생쥐에 장기간 서구식 사료를 급여했을 때 내인성 SHP 단백질의 수준이 증가함을 보고하였다. 또한 HepG2 세포배양을 통한 실험에서, CDCA 처리가 내인성 SHP 단백질의 수준을 증가시킬 뿐만 아니라 외인성으로 발현된 flag-SHP의 분해율을 감소시켰다. 그리고 HepG2 세포를 ad-flag-SHP로 유전자 형질전환 시켰을 때, 담즙산에 의해 유도되어진 소장 FGF-19이 외인성으로 발현된 flag-SHP 단백질의 반감기를 증가시켰다. 그러나 cholic acid와 FGF-19에 의한 내인성 SHP 단백질의 발현수준과 분해율은 생쥐 또는 배양된 간암세포주에서 아직 명확히 이해되고 있지 않다. 이 연구는 cholic acid의 처리가 생쥐에서 내인성 SHP 단백질의 수준에 미치는 영향과, FGF-19이 HepG2 세포주에서 내인성 SHP 단백질의 분해율에 미치는 영향을 조사하였다. 정상적인 사료를 급여한 대조군 생쥐에서의 내인성 SHP 단백질 수준과 비교하여, 0.5%의 cholic acid를 첨가한 사료를 급여한 생쥐에서는 12시간과 24시간의 처리기간 동안에 내인성 SHP 단백질의 수준이 증가하였다. 배양된 인간 간암세포주인 HepG2에서 CDCA의 처리는 CDCA를 처리하지 않은 대조군 세포주와 비교하여 내인성 SHP 단백질의 분해율을 유의성 있게 변화시키지 않았다. 한편 외인성 ad-flag-SHP 단백질에 대한 이전의 연구와 일치하게, HepG2 세포에 cyclohexamide를 처리하였을 때 FGF-19는 내인성 SHP 단백질의 분해율을 현저히 감소시켰다. 이러한 결과는 담즙산과 FGF-19 모두 생쥐의 간과 HepG2 세포주에서 내인성 SHP 단백질의 수준을 증가시킴을 제시한다.

월비탕(越婢湯)이 고지방식이(高脂肪食餌)로 유도된 비만 생쥐에 미치는 영향 (Anti-obesity Effects of Wolbi-tang(越婢湯) on the Obese-mice Induced by High-fat Diet)

  • 박지현;홍서영
    • 한방재활의학과학회지
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    • 제21권2호
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    • pp.31-48
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    • 2011
  • Objectives : In order to investigate the anti-obesity effects of Wolbi-tang(here in after referred to WBT) on the obese gene and obese inhibitory, C57BL/6 mice were induced by high-fat diet. Methods : C57BL/6 mice were divided into 5 groups(normal, only high-fat diet, high-fat diet with Reductil, high-fat diet with WBT 400, 200 mg/kg extract) and fed for 5 weeks. And observed body weight change, total cholesterol, low density lipoprotein cholesterol(LDL-cholesterol), high density lipoprotein cholesterol (HDL-cholesterol), triglyceride, glucose, leptin change, alanine transaminase(ALT), aspartate transaminase(AST), serum creatinine, the expression of ${\beta}3$-adrenergic receptor(${\beta}3AR$), leptin, uncoupling protein(UCP2) gene in 3T3-L1 adipocyte, 3T3-L1 adipocyte proliferation, histological analysis of adipose tissue and liver tissue. Results : 1. Refer to cell cytotoxicity, viability of human fibroblast cells(hFCs) showed not significant changes. 2. The amount of ALT, AST was decreased significantly in WBT 400 mg/kg, 200 mg/kg groups. The amount of creatinine showed not significant changes. 3. Body weight was decreased significantly in WBT 400 mg/kg, 200 mg/kg groups. 4. The amount of total cholesterol and triglyceride was decreased significantly in WBT 400 mg/kg, 200 mg/kg groups. LDL-cholesterol was decreased and HDL-cholesterol was increased significantly in WBT 400 mg/kg groups. 5. The amount of glucose was decreased significantly in WBT 400 mg/kg groups. 6. The amount of serum leptin was decreased significantly in WBT 400 mg/kg, 200 mg/kg groups. 7. The revelation of ${\beta}3AR$ in 3T3-L1 adipocyte was increased significantly in WBT $100{\mu}g/ml$, $50{\mu}g/ml$ groups. The revelation of leptin was decreased significantly in WBT $100{\mu}g/ml$, $50{\mu}g/ml$ groups. The revelation of UCP2 was decreased significantly in WBT $100{\mu}g/ml$ group. 8. 3T3-L1 adipocyte proliferation was decreased significantly in WBT $100{\mu}g/ml$, $50{\mu}g/ml$ groups. The size of adipocyte was decreased relative to the control group in WBT 400 mg/kg group. 9. The adipose vacuoles in liver tissue was decreased relative to the control group. Conclusions : These results suggested that WBT has inhibitory effects of obesity. WBT might be applicated on treatment of obesity and metabolic syndrome. Further studies analysing its effects were needed.

Collagen으로 유발된 생쥐의 관절염에 대한 당귀사역가오수유생강탕(當歸四逆加吳茱萸生薑湯)의 효과 (Effects of Danggwisayeokgaohsuyusaenggang-tang(當歸四逆加吳茱萸生薑湯) on Collagen-induced Arthritis in Mice)

  • 김민균;오민석
    • 한방재활의학과학회지
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    • 제21권2호
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    • pp.63-85
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    • 2011
  • Objectives : This study was carried out to know the effects of Danggwisayeokgaohsuyusaenggang-tang(hereinafter referred to DST) on arthritis induced by collagen on DBA/1 OlaHsd mice. Methods : For this purpose, DST was orally administered to mouse with arthritis induced by collagen II. Cytotoxicity, high performance liquid chromatograph(HPLC) analysis, arthritis index, value of immunocyte in draining lymph node and paw joint, cytokine were measured in vivo. Results : 1. The cytotoxicity against human fibroblast cells(hFCs) was not measured in any concentration. 2. In HPLC analysis, There are high peak patterns at 8 minute(min), 12 min, 35 min, 45 min. 3. The arthritis index was decreased significantly. 4. The degree of arthritis induced damage of joint of DST group is slight compared with control group in histopathologic observation(Hematoxylin and eosin stain(H&E), Masson's trichrome(M-T) staining). 5. In total cell counts of draining lymph node(DLN) and paw joint, the cells in DLN decreased significantly on DST 200 mg/kg and the cells in paw joint decreased significantly on 200 mg/kg and 50 mg/kg. 6. In DLN, $CD4^+/CD25^+$, $CD3^+/CD69^+$, major histocompatibility complex(MHC), class-II/$CD11c^+$ cells decreased significantly on DST 200 mg/kg and 50 mg/kg $CD3^+/CD8^+$ cells decreased significantly on DST 200 200 mg/kg, $CD4^+$, $CD3^+/CD44^+$ cells decreased. 7. In paw joints, $CD4^+$, $CD11b^+/Gr-1^+$ cells decreased significantly on DST 200 mg/kg and 50 mg/kg. 8. In joints, levels of $IL-1{\beta}$, IL-6, $TNF-{\alpha}$, cyclo-oxygenase-2(COX-2), NOS-II were decreased on DST 200 mg/kg and DST 50 mg/kg. 9. In analysing of cytokine in CD3/CD28 activated spleen, IL-17 was decreased significantly, IL-4 was increased significantly $INF-{\gamma}$ was decreased on DST 200 mg/kg. 10. In analysing of cytokine in collagen activated spleen, IL-17 were decreased significantly, IL-4 was increased significantly. Conclusions : This results demonstrated that DST suppressed the inflammatory progression of collagen-induced arthritis(CIA) mice and supported further studies are required to survey continuously in looking for the effective substance and mechanism in the future.

치주인대섬유아세포가 파골세포분화에 미치는 영향 (Human Periodontal Ligament Fibroblasts Support the Osteoclastogenesis of RAW264.7 Cells)

  • 이호;전용선;최승환;김형섭;오귀옥
    • Journal of Periodontal and Implant Science
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    • 제32권4호
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    • pp.733-744
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    • 2002
  • The fibroblasts are the principal cells in the periodontal ligament of peridontium. As the periodontal ligament fibroblasts (PDLF) show similar phenotype with osteoblasts, the PDLF are thought to play an important role in alveolar bone remodeling. Cell-to-cell contacted signaling is crucial for osteoclast formation. Recently it has been reported that PDLJ enhance the bone resorbing activity of osteoclasts differentiated from hematopoietic preosteoclasts. The aims of this study were to $clarify\;^{1)}$ the mechanism of PDLF-induced osteoclastogenesis $and\;^{2)}$ whether we can use preosteoclast cell line instead of primary hematopoietic preosteoclast cells for studying the mechanism of PDLF-induced osteoclastogenesis. Osteoclastic differentiation of mouse macrophage cell line RAW264.7 was compared with that of mouse bone marrow-derived M-CSF dependent cell (MDBM), a well-known hematopoietic preosteoclast model, by examining, 1) osteoclast-specific gene expression such as calcitonin receptor, M-CSF receptor (c-fms), cathepsin K, receptoractivator nuclear factor kappa B (RANK) ,2) generation of TRAP(+) multinucleated cells (MNCs), and 3) generation of resorption pit on the $OAAS^{TM}$ plate. RAW264.7 cultured in the medium containing of soluble osteoclast differentiation Factor (sODF) showed similar phenotype with MDBM-derived osteoclasts, those are mRNA expression pattern of osteoclast-specific genes, TRAP(+) MNCs generation, and bone resorbing abivity. Formation of resorption pits by osteoclastic MNCs differentiated from sODF-treated RAW264.7, was completely blocked by the addition of osteoprotegerin (OPG), a soluble decoy receptor for ODF, to the sODF-containing culture me야um. The effects of PDLF on differentiation of RAW264.7 into the TRAP(+) multinucleated osteoclast-like cells were examined using coculture system. PDLF were fxed with paraformaldehyde, followed by coculture with RAW264.7, which induced formation of TRAP(+) MNCs in the absence of additional treatment of sODF. When compared with untreated and fixed PDLF (fPDLF), IL-1 ${\beta}$-treated, or lipopolysaccha-ride-treated and then fixed PDLF showed two-folld increase in the supporting activity of osteoclastogenesis from RAW264.7 coculture system. There were no TRAP(+) MNCs formation in coculture system of RAW264.7 with PDLF of no fixation. These findigs suggested that we can replace the primary hematopoietic preosteoclasts for RAW264. 7 cell line for studying the mechanism of PDLF-induced osteoclastogenesis, and we hypothesize that PDLF control osteoclastogenesis through ODF expression which might be enhanced by inflammatory signals.

Molecular characterization and expression of a disintegrin and metalloproteinase with thrombospondin motifs 8 in chicken

  • Lee, Ra Ham;Lee, Seokhyun;Kim, Yu Ra;Kim, Sung-Jo;Lee, Hak-Kyo;Song, Ki-Duk
    • Asian-Australasian Journal of Animal Sciences
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    • 제31권8호
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    • pp.1366-1372
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    • 2018
  • Objective: A disintegrin and metallopeptidase with thrombospondin motifs type 8 (ADAMTS8) is crucial for diverse physiological processes, such as inflammation, tissue morphogenesis, and tumorigenesis. The chicken ADAMTS8 (chADAMTS8) gene was differentially expressed in the kidney following exposure to different calcium concentrations, suggesting a pathological role of this protein in metabolic diseases. We aimed to examine the molecular characteristics of chADAMTS8 and analyze the gene-expression differences in response to toll-like receptor 3 (TLR3) stimulation. Methods: The ADAMTS8 mRNA and amino acid sequences of various species (chicken, duck, cow, mouse, rat, human, chimpanzee, pig, and horse) were retrieved from the Ensembl database and subjected to bioinformatics analyses. Reverse-transcription polymerase chain reaction (RT-PCR) and quantitative PCR (qPCR) experiments were performed with various chicken tissues and the chicken fibroblast DF-1 cell line, which was stimulated with polyinosinic-polycytidylic acid (poly[I:C]; a TLR3 ligand). Results: The chADAMTS8 gene was predicted to contain three thrombospondin type 1 (TSP1) domains, whose amino acid sequences shared homology among the different species, whereas sequences outside the TSP1 domains (especially the amino-terminal region) were very dif­ferent. Phylogenetic analysis revealed that chADAMTS8 is evolutionarily clustered in the same clade with that of the duck. chADAMTS8 mRNA was broadly expressed in chicken tissues, and the expression was significantly up-regulated in the DF-1 cells in response to poly(I:C) stimulation (p<0.05). These results showed that chADAMTS8 may be a target gene for TLR3 signaling. Conclusion: In this report, the genetic information of chADAMTS8 gene, its expression in chicken tissues, and chicken DF-1 cells under the stimulation of TLR3 were shown. The result suggests that chADAMTS8 expression may be induced by viral infection and correlated with TLR3-mediated signaling pathway. Further study of the function of chADAMTS8 during TLR3-dependent inflammation (which represents RNA viral infection) is needed and it will also be important to examine the molecular mechanisms during different regulation, depending on innate immune receptor activation.

노니 추출물의 주름개선 효과연구 (Anti-wrinkle Effect of Morinda citrifolia (Noni) Extracts)

  • 이정노;김상우;유영경;이강태;이건국
    • 대한화장품학회지
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    • 제32권4호
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    • pp.227-231
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    • 2006
  • 주름생성의 주요 원인은 자외선, 공기 오염, 담배 연기, 스트레스 등을 들 수 있다. 특히, 자외선은 피부에 다양한 형태로 영향을 주어 깊은 주름, 잔주름, 피부거침, 피부건조와 같은 현상을 발생시켜 피부노화를 유발시킨다. 이러한 주름 생성 해결 및 피부 노화 문제의 해결은 화장품에서 가장 활발하게 연구되는 분야 중 하나이다. 최근에 열대 식물인 노니로부터 추출한 유효성분이 주름 개선에서 가장 중요한 콜라겐 생합성을 촉진하는 효과를 확인하였으며, 특히 노니 추출물로부터 분리한 스코폴레틴(scopoletin)이 이러한 효과를 주는 활성 성분임을 확인하였다. 스코폴레틴은 인체 섬유아 세포를 이용한 콜라겐 합성 촉진 효과 시험에서 농도 의존적으로 콜라겐 합성을 촉진하는 활성을 보였다($0.2{\mu}g/mL-89.5%$ 콜라겐 생합성 촉진. 또한, 주름 개선 효과를 확인하기 위해 3% (v/v) 노니 추출물을 함유한 크림을 이용하여 12주 동안 임상연구를 수행하여, 안면의 주름 감소 정도를 측정하였으며 이 제품이 주름을 감소시켜주는 뛰어난 효과가 있다는 것을 확인하였다. 이러한 결과들은 노니 추출물이 주름 개선효과가 매우 높으며, 노화 방지 원료로서 이용 가능성이 매우 높다는 것을 보여주는 것이라 할 수 있다.