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ATO-ALL이 인간유래 피부 각질형성세포인 HaCaT 세포에서 피부재생에 미치는 효과
신윤민 ( Younmin Shin ),이원길 ( Won-kil Lee ),김승형 ( Seung Hyung Kim ),최정준 ( Jeong June Choi ) 대전대학교 한의학연구소 2019 한의학연구소 논문집 Vol.28 No.2
Objectives : Atopic dermatitis is an irritable skin disease accompanying rash and itching leading to impaired skin barrier. ATO-ALL is an ethanol extract of natural products comprising 12 herbs and effective on atopic dermatitis. In this study, we aimed to propose that the effect of ATO-ALL on skin regeneration in human keratinocyte cell line, HaCaT cells. Methods : To evaluate the skin regenerating effects of ATO-ALL, scratch wound healing assay, bromodeoxyuridine (BrdU) assay, and propidum iodide (PI) assay were performed using cultured HaCaT cell line. Result : Scratch wound healing assay showed that ATO-ALL was able to enhance the gap filling activity more than 2-fold at 7 ppm concentration compared with control group. BrdU assay demonstrated that ATO-ALL treatment increased the de novo cell proliferation in a dose-dependent manner. Finally, PI assay indicated that the cell cycle of HaCaT cells was modulated by ATO-ALL treatment. Conclusions : These results suggested that ATO-ALL may have skin regenerating effects by increasing cell proliferation via cell cycle regulation. Taken together, ATO-ALL is supposed to have a potential on regeneration of damaged skin or functional disease including atopic dermatitis.
TNCB로 유발한 아토피피부염 생쥐 모델에서 천일염가미방(天日鹽加味方)과 청기해독산(淸肌解毒散)의 병용 효과
이경미 ( Kyoung Mee Lee ),김선빈 ( Seon Bin Gim ),최학주 ( Hak Ju Choi ),최정준 ( Jeong June Choi ),노성수 ( Seong Soo Roh ),김동희 ( Dong Hee Kim ) 대한본초학회 2009 大韓本草學會誌 Vol.24 No.4
Objectives: Atopic dermatitis is a chronic inflammatory disease characterized by typically distributed eczematous skin lesion with pruritus, lichenification and dry skin. In this study, we performed to assess the therapeutic effects of co-treatment of Chenilyeomgamibang (CGB) and Chenggihaedok-san (CHS, C&C) on the TNCB(trinitrochlorobenzene)-induced atopic dermatitis in NC/Nga mice, characterized by the onset of atopic dermatitis along with an increase the number of inflammatory cells and dysregulation of Th2 cytokines. Methods: Defined amount of CGB was sprayed on mice skin and CHS was simultaneously orally administrated to TNCB treated NC/Nga mice for 5 weeks. The immune cell types were caracterized by flow cytometry using each specific antibody. The amount of Th2 cytokines in serum and splenocytes culture supernatant were measured by ELISA. Results: Administration of C&C significantly reduced clinical dermatitis severity including pruritus, edema, eczematous and erythema. Histological findings indicated that the thickening of epidermis/ dermis and dermal infiltration of inflammatory cells were dramatically reduced. Flow cytometry analysis showed that infiltrated immune cell numbers of CCR3+, B220+/IgE+, Gr-1+/CD11b+, and CD117+ were significantly reduced in C&C-treated dorsal skin lesion. Furthermore, T cell composition rate in PBMC was also dramatically decreased by the treatment. C&C greatly down-regulated production of Th2 cytokines including IL-4, IL-5, IL-13 in the serum. The down-regulatory effects of C&C on these Th2 cytokines production were also detected in CD3/ CD28 activated splenocytes. Conclusions: These results indicated that C&C is a plausible therapeutic agent for treatment of atopic dermatitis through regulating the Th2 skewed immune system.
HPLC 를 이용한 가는장구채 추출물의 Isoscoparin 분석법 개발
권진관 ( Jin Gwan Kwon ),서찬곤 ( Changon Seo ),정연우 ( Yeon Woo Jung ),최용문 ( Yongmun Choi ),신현탁 ( Hyun Tak Shin ),정수영 ( Su Young Jung ),최정준 ( Jeong June Choi ),김진규 ( Jin Kyu Kim ) 대한화장품학회 2021 대한화장품학회지 Vol.47 No.1
본 연구는 가는장구채(Silene seoulensis) 추출물을 기능성 화장품소재로 개발하기 위해 isoscoparin을 지표성분으로 선정하고, 품질관리를 위해 high performance liquid chromatography (HPLC)를 이용하여 분석법을 개발하였다. 분석용 칼럼은 Unison US-C<sub>18</sub>을, 이동상은 0.05% (v/v) trifluoroacetic acid (TFA)와 메탄올을 사용하여 컬럼 온도는 35 ℃ 에서 유속 1.0 mL/min 로 검출파장은 330 nm에서 검출하였다. International conference on harmonization (ICH) 가이드라인(version 4, 2005)을 근거로 하여 특이성, 직선성, 정밀성, 정확성, 검출한계 및 정량한계를 분석하여 분석방법을 검증하였다. 분석결과, 검출한계 및 정량한계는 0.02 및 0.07 mg/mL로 나타났으며, 검량곡선은 상관계수 값이 0.99988로 양호한 직선성을 보였고 정밀성 분석결과도 0.46% 이하로 확인되었다. 또한, 회수율은 99.10 ~ 101.61% 범위로 정확성이 있음을 확인하였다. 따라서, 본 분석법은 가는장구채 추출물 지표성분의 분석법으로 적합한 시험법임이 검증되었다. In this study, isoscoparin was selected as an indicator component to develop Silene seoulensis extract as a functional cosmetic material, and we developed an analysis method using high performance liquid chromatography (HPLC) for quality control. HPLC was performed on a Unison US-C<sub>18</sub> with a gradient elution of 0.05% (v/v) trifluoroacetic acid (TFA) and methanol at a flow rate of 1.0 mL/min at 35 ℃, and the detection wavelength was 330 nm. The HPLC method was performed in accordance with the international conference on harmonization (ICH) guideline (version 4, 2005) of analytical procedures with respect to specificity, precision, accuracy, and linearity. The limits of detection and quantitation were 0.02 and 0.07 mg/mL respectively. Calibration curves showed good linearity (R2 > 0.99988), and the precision of analysis was satisfied (less than 0.46%). In addition, the recovery rate was in the range of 99.10 to 101.61%, it was shown to be accurate. This result indicated that the established HPLC method is very useful for the determination of marker compounds in Silene seoulensis extracts.
HPLC를 이용한 노루오줌 추출물의 Avicularin 및 Quercitrin 분석법 개발
권진관 ( Jin Gwan Kwon ),정연우 ( Yeon Woo Jung ),서찬곤 ( Changon Seo ),홍성수 ( Seong Su Hong ),신현탁 ( Hyun Tak Shin ),정수영 ( Su Young Jung ),최정준 ( Jeong June Choi ),김진규 ( Jin Kyu Kim ) 대한화장품학회 2019 대한화장품학회지 Vol.45 No.4
본 연구는 노루오줌(Astilbe chinensis) 추출물을 기능성 화장품소재로 개발하기 위해 avicularin, quercitrin을 지표성분으로 선정하고, 품질관리를 위해 high performance liquid chromatography (HPLC)를 이용하여 분석법을 개발하였다. 분석용 칼럼은 Capcell Pak C<sub>18</sub> MGII을, 이동상은 0.05% (v/v) trifluoroacetic acid (TFA)와 아세토니트릴을 사용하여 컬럼 온도는 30℃ 에서 유속 1.0 mL/min 로 검출파장은 254 nm에서 검출하였다. International conference on harmonization (ICH) 가이드라인(version 4, 2005)을 근거로 하여 특이성, 직선성, 정밀성, 정확성, 검출한계 및 정량한계를 분석하여 분석방법을 검증하였다. 분석결과, 검출한계 및 정량한계는 avicularin 0.094 및 0.285 mg/mL, quercitrin 0.031 및 0.095 mg/mL로 나타났으며, 검량곡선은 상관계수 값이 avicularin 0.99990, quercitrin 0.99994로 양호한 직선성을 보였고 정밀성 분석결과도 avicularin 0.59%, quercitrin 0.63% 이하로 확인되었다. 또한, 회수율은 avicularin 100.97 ∼ 101.77%, quercitrin 100.18 ∼ 100.32% 범위로 정확성이 있음을 알 수 있다. 따라서, 본 분석법은 노루오줌 추출물 지표성분의 분석법으로 적합한 시험법임이 검증되었다. This study attempted to eatablish a high performance liquid chromatography (HPLC) analysis method for the determination of avicularin, quercitrin as a part of the quality control for the development of functional cosmetic materials from Astilbe chinensis extract. HPLC was performed on a Capcell Pak C<sub>18</sub> MGII column with a gradient elution of 0.05% (v/v) trifluoroacetic acid (TFA) and acetonitrile at a flow rate of 1.0 mL/min at 30 ℃. The analyte was detected at 254 nm. The HPLC method was performed in accordance with the international conference on harmonization (ICH) guideline (version 4, 2005) of analytical procedures with respect to specificity, precision, accuracy, and linearity. The limits of detection and quantitation of avicularin and quercitrin were 0.094 and 0.285 mg/mL, 0.031 and 0.095 mg/mL respectively. Calibration curves showed good linearity r<sup>2</sup> > 0.99990 for avicularin and r<sup>2</sup> > 0.99994 for quercitrin. Precision of analysis was satisfied with less than 0.59% for avicularin and 0.63% for quercitrin. Recoveries of quantified compounds ranged from 100.97 to 101.77% for avicularin and 100.18 to 100.32% for quercitrin. These result indicated that the established HPLC method is very useful for the determination of marker compounds in A. chinensis extracts.