• 대한치과교정학회지
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• 제9권1호
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• pp.9-14
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• 1979

The purpose of this study was to pursue the biosynthesis of proteins of human and bovine periodontal ligaments in vitro system. The excised periodontal ligaments from human and bovine were incubated in Krebs-glucose medium containing $^3H$-proline. After incubation the incubated periodontal ligaments were homogenized and the proteins were treated with 0.1%sodium dodecyl sulfate and $\beta$-mercaptoethanol. Separation of the protein fractions was performed with agarose gel column chromatography and SDS acrylamide gel electrophoresis. The results indicated as follow: 1. Only a small percentage of $^3H$-proline incorporated into proteins was hydroxylated to $^3H$-hydroxyproline. 2. The labeled proteins in periodontal ligaments showed a wide distribution of molecular weight. But only small amounts of labeled protein were found that were characteristics of the molecular weight of collagen. 3. In all of the combined fractions of gel filtration, the degree of hydroxylation was small.

• 대한한방부인과학회지
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• 제25권1호
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• pp.20-33
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• 2012

Objectives: This study was performed to elucidate the effects of Kanghwalsokdan-tang extract(KS) on hyper-plasy of collagen and cell damage in UVB-irradiated dermal fibroblast. Methods: To demonstrate the effects of KS on wound healing we used human dermal fibroblast(F6). We evaluated the amount of increased PICP, TIMP-1 in dermal fibroblast. PICP, TIMP-1 concentration was measured using EIA kit. Also, we measured the nitrite production, and LDH release in UVB-irradiated dermal fibroblast to elucidate the action-mechanism of KS. Results: 1. KS decreased the cell proliferation of dermal fibroblast. 2. KS decreased the biosynthesis of collagen in dermal fibroblast. 3. KS decreased the synthesis of TIMP-1 in dermal fibroblast. 4. KS had no effect on the LDH-release of UVB-irradiated dermal fibroblast. 5. KS inhibited nitrite production in UVB-irradiated dermal fibroblast. Conclusions: From the results, we concluded that KS has a protective effect on wound healing and photoaging.

• 대한화장품학회지
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• 제25권2호
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• pp.59-75
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• 1999

Skin is continuously exposed to external stimuli including ultraviolet radiation, which is a major cause of skin photoaging. According to recent discoveries, UVA with a lower energy but deep-penetrating properties, compared to UVB, is likely to play a major part in causing skin photoaging. The clinical and histochemical changes of photoaging are well characterized, but the biochemical mechanisms are poorly understood partly due to the lack of suitable experimental systems. In this work, three-dimensional, reconstituted skin culture models were prepared. After certain period of maturation, the equivalent models were shown to be similar in structure and biochemical characteristics to normal skin. Mature dermal and skin equivalent models were exposed to sub-lethal doses of UVA, and the effects of UVA relevant to dermal photoaging were monitored, including the production of elastin, collagen, collagenase(MMP-1), and tissue inhibitor of metalloproteinases-1 (TIMP-1). Interestingly, dermal and skin equivalents reacted differently to acute and chronic exposure to UVA. Elastin production was increased as soon as one week after commencing UVA irradiation by chronic exposure, although a single exposure failed to do so. This early response could be an important advantage of equivalent models in studying elastosis in photoaged skin. Collagenase activity was increased by acute UVA irradiation, but returned to control levels after repeated exposure. On the other hand, collagen biosynthesis, which was increased by a single exposure, decreased slightly during 5 weeks of prolonged UVA exposure. Collagenase has been thought to be responsible for collagen degeneration in dermal photoaging. However, according to the results obtained in this study, elevated collagenase activity is not likely to be responsible for the degeneration of collagen in dermal photoagig, while reduced production of collagen may be the main reason. It can be concluded that reconstituted skin culture models can serve as useful experimental tools for the study of skin photoaging. These culture models are relatively simple to construct, easy to handle, and are reproducible Moreover the changes of dermal photoaging can be observed within 1-4 weeks of exposure to ultraviolet light compared to 4 months to 2 years for human or animal studies. These models will be useful for biochemical and mechanistic studies in a large number of fields including dermatology, toxicology, and pharmacology.

• Journal of Ginseng Research
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• 제31권2호
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• pp.86-92
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• 2007

본 실험은 홍삼을 이용하여 주름 예방에 효과가 있는 건강 기능 식품을 개발하기 위한 기초 자료로 활용하고자 홍삼이 사람의 섬유아세포에 대한 콜라겐 생합성 정도, 콜라겐 분해효소 활성, 세포 증식, 항산화 활성에 미치는 영향을 조사하였다. 조사포닌은 홍삼물엑기스 (WE), 에탄올 엑기스 (EE), 에탄올엑기스를 88% 주정으로 용해시켜 정제한 엑기스 (PE) 에서 각각 72 mg/g, 107 mg/g, 220 mg/g으로 함유되어 있었다. 홍삼시료는 1 ${\mu}g/ml$, 5 ${\mu}g/ml$, 10 ${\mu}g/ml$의 농도로 첨가하여 섬유아세포를 배양하였다 . 콜라겐 생합성 정도는 대조군이 1.86 ${\mu}g/ml$이었고 Retinoic acid의 경우 0.03 ${\mu}g/ml$의 농도로 처리했을 때 2.85 ${\mu}g/ml$이었다 . 홍삼의 첨가농도 10 ${\mu}g/ml$을 기준으로 할 때 물엑기스에서는 대조군과 유사하였고, EE (ethanol extract)에서는 2.05 ${\mu}g/ml$, PE (purified ethanol extract)에서는 2.58 ${\mu}g/ml$, ginsenoside $Rg_1$은 2.01 ${\mu}g/ml$이었으며 $Rb_1$은 3.07 ${\mu}g/ml$의 콜라겐을 생성하여 대조군에 비하여 165% 증가하였으며 retinoic acid보다 더 높았다. MMP-1 활성에서는 10 ${\mu}g/ml$의 농도로 배양했을 때 EE, PE, $Rg_1$, $Rb_1$에서 대조군의 92%, 94%, 91%, 48%을 나타내어 $Rb_1$이 저해효과가 뚜렷하였다. MTT assay에서 모든 시료는 대조군에 비하여 84%${\sim}$96%의 수준으로 세포 생존율을 나타내어 의미있는 세포 독성을 나타내지 않았다. 항산화 정도를 나타내는 SOD 활성은 높은 수준은 아니었으나 Vit.C에 비하여 28${\sim}$79%의 활성을 나타내었다. 이상의 실험 결과를 종합해 볼 때 홍삼 시료는 사람의 섬유아세포에 대하여 콜라겐 생합성을 증가시키고 MMP-I 활성을 억제하였으며 항산화 효과를 나타내어 피부 주름 생성을 억제하는 효과가 있는 것으로 확인되었고, 홍삼의 사포닌 성분 중에서는 $ginsenoside-Rg_1$에 비하여 $ginsenoside-Rb_1$이 더 효과적이었다.

• Journal of Ginseng Research
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• 제31권4호
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• pp.196-202
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• 2007

본 실험은 홍삼, 사상자, 산수유를 이용하여 주름 예방 효과가 있는 건강기능 식품을 개발하기 위한 기초자료로 활용하기 위하여 개별추출물과 이들의 혼합물 WM1, WM3 (KTNG0345)에 대하여 사람의 섬유아세포를 이용한 세포증식, 콜라겐 생합성 정도, MMP-1 효소활성, 항산화 활성을 조사하였다. 사용된 시료는 $1{\sim}10\;{\mu}g/ml$의 농도로 첨가하여 세포를 배양했을 때 $83{\sim}100%$의 세포증식율을 나타내어 세포 독성을 나타내지 않았으며, 콜라겐 생합성 정도는 대조군이 474.8 ng/ml이었고 retinoic acid는 568.3 ng/ml이었으며, 5 ug/ml의 농도를 기준으로 할 때 홍삼은 533.9ng/ml, 사상자는 539.3 ng/ml, 산수유는 514.1 ng/ml이었다. 홍삼과 생약을 68:232의 비율로 혼합한 WM1은 474.6 ng/ml이었고 홍삼과 생약을 136:164의 비율로 혼합한 WM3는 561.4 ng/ml로 증가하였다. MMP-1활성은 10 ug/ml의 농도에서 홍삼(ER)은 31.9 ng/ml, 사상자(WT)는 32.85 ng/ml, 산수유(WC)는 32.0 ng/ml, WM1은 31.3 ng/ml이었고 WM3(KTNG0345)는 28.85 ng/ml으로 대조군의 88% 수준으로 감소하였다. SOD활성에서는 1000 ug/ml의 농도로 처리하였을 때 65-97% 항산화활성을 나타내었다. 이들의 결과를 종합해 보면 홍삼과 사상자, 산수유 및 이들 혼합물은 human dermal fibroblas에 대하여 세포 독성을 나타내지 않으면서 콜라겐 생합성을 촉진하고 MMP-1의 활성을 억제하였으며 홍삼과 생약 엑기스를 136:164의 비율로 혼합했을 때 그 효과가 증진되었다. WM3 (KTNG0345)는 지표성분으로서 ginsenoside-$Rb_1$, torilin, loganin을 각각 10.85 mg/g, 0.128 mg/g, 3.92 mg/g 함유하였다.

• 한국축산식품학회지
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• 제35권1호
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• pp.58-70
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• 2015

This study was conducted to examine the effect of egg shell membrane hydrolysates (ESMH) on wrinkle, UV, and moisture protection for cosmetic use. ESMH were fragmented as whole ESMH (before fractioning), Fraction I (> 10 kDa), Fraction II (3-10 kDa), and Fraction III (< 3 kDa). In order to test whether fractionated ESMH can be used for functional cosmetic materials, we examined not only the level of hyaluronic acid and collagen production, but also the MMP-1 activity using a HaCaT and CCD-986Sk cell line. Our study treated each sample of fractionated ESMH with different concentrations (0.01, 0.1, 1 mg/mL). In our in vivo research, we used hairless mice that had been exposed to UV-B to induce wrinkles for 7 wk, then applied Fraction I to the treatment group for 5 wk and then tested skin thickness, minimum erythema dose and moisture content. In addition, Fraction I was high in collagen and HA biosynthesis and it was better than TGF-${\beta}$ in improving of the skin. When TNF-${\alpha}$ caused MMP-1 activity in the CCD-986Sk cells, the whole ESMH and Fraction I proved to be effective in hindering the induction of collagenase depending on the concentration, and also showed outstanding effects in the suppression of skin aging. We found that the treatment group mice's UV-B radiation-induced skin damage was largely mitigated compared to that of the non-treatment group mice. Thus, we have concluded that EMSH helps to mitigate UV-B radiation-induced wrinkles, collagen, HA, MMP-1 activity and can be used for functional cosmetic materials.

• 한국수산과학회지
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• 제41권1호
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• pp.7-12
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• 2008

To develop a whitening agent, cytotoxicity of the soluble collagen isolated from Todarodes pacificus (CIT) was evaluated. CIT tested for cytotoxicity on human dermal fibroblast (CCD-986sk) was exhibited very low cytotoxicity. Because tyrosinase is the key enzyme for melanin biosynthesis, the use of various tyrosinase inhibitors is a common practice for whitening purpose in cosmetics. Tyrosinase inhibitory activity and melanin production assay were measured to confirm the whitening effect. The inhibitory effect of MMP-1 in UV-irradiated human dermal fibroblast was also performed. CIT showed strong inhibition potency on tyrosinase by 51.5% at 0.2 mg/mL which increased the inhibition by increasing the concentration of CIT, and showed 69.1% inhibition at 1.0 mg/mL. CIT showed strong inhibition effect on melanin production with 82% at 1.0 mg/mL. The CIT also reduced about 76% expression of MMP-1 in UV-irradiated CCD-986sk cell at 1.0 mg/mL. From the preliminary observations, we suggest that the collagen isolated from CIT could be a potential source of natural skin-whitening and anti-aging agents for the photo-damaged skin.

• 한국자원식물학회지
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• 제29권2호
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• pp.155-162
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• 2016

본 연구에서는 Dendrobium speciosum 에탄올추출물의 티로시나아제 억제를 통한 멜라닌형성 저해 효과 및 주름개선효과에 대해 연구하였다. Dendrobium speciosum추출물은 α-MSH만 처리한 군에 비해 티로시나아제 활성을 농도 의존적으로 저해시킴으로서 멜라닌 함량을 감소시켰다. 더 자세한 메카니즘을 알아보기 위하여 멜라닌 합성 관련 유전자인 Tyrosinase-related protein-1, tyrosinase-related protein-2, Microphthalmia-associated transcription factor 발현을 확인하였을 때 Dendrobium speciosum처리군에서 유전자 발현이 농도 의존적으로 감소하였다. 또한 Dendrobium speciosum처리 시 type I procollagen 합성이 증가함으로써 Dendrobium speciosum처리가 collagen 생합성을 증대시킬 것으로 사료된다. 이 결과를 바탕으로 Dendrobium speciosum에탄올 추출물 처리 시 멜라닌합성 관련 유전자의 발현 및 티로시나아제 활성을 저해시켜 멜라닌 합성이 감소됨에 따라 효과적인 미백활성을 가지며 collagen합성을 증대시킴으로서 주름개선에 탁월한 효과를 가지는 것으로 사료되며 이는 기능성 화장품 소재로 사용가능할 것으로 사료된다.

• 생약학회지
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• 제35권4호통권139호
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• pp.315-319
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• 2004

Three coumarin derivatives, which stimulate collagen biosynthesis in human fibroblasts were isolated from Angelica dahurica root which has been used in traditional medicine for cold, headache, discharging blood and women's diseases. Three coumarin derivatives were identified as 5-benzofuranacrylic acid, $6,7-dihydroxy-4-(3-methyl-2-butenyl)-{\delta}-lactone\;(prangenidin)$, 5-benzofuranacrylic acid, $6,7-dihydroxy-4-methoxy-{\delta}-lactone\;(8-hydroxybergapten)$ and 5-benzofuranacrylic acid, 8-hydroxypsoralen(xanthotoxol) from their spectral data.

• 대한한방부인과학회지
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• 제29권2호
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• pp.66-82
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• 2016

Objectives : This study was conducted to evaluate the effect of Kanghwalsokdan-tang Gamibang water extract (KSG) on osteoporosis. Methods : RANKL-stimulated RAW 264.7 was used to evaluate inhibitory effect of KSG osteoclast differentiation and gene expression. We counted TRAP (+) multinucleated cells and measured TRAP activity and mRNA expressions of osteoclastogenesis-related genes (NFATc1, MITF, JNK1, cathepsin K, MMP-9) to figure out the effect of KSG on osteoclast. Osteoblastogenesis was also determined in rat calvarial cell. Alkaline phosphatase (ALP) activity, bone matrix protein and collagen synthesis were measured by using murine calvarial cell. Results : KSG inhibited the differentiation of osteoclast precursor cell and expression of genes related osteoclastogenesis like NAFTc1, MITF, c-fos, JNK1, Cathepsin K, MMP-9 and TRAP. KSG increased cell division and function of osteoblast separated from the skull of a rat and ALP synthesis, biosynthesis of bone matrix protein and collagen. Conclusions : Reviewing these results, KSG has efficacy on osteoclast inhibition and osteoblast activation. After further study, KSG will be able to apply for osteoporosis treatment and prevention.