상황버섯(Phellinus Linteus) 추출물 분획들의 암세포에 대한 세포독성 및 항산화효과 연구

김형자 ( Hyoung Ja Kim ),진창배 ( Chang Bae Jin ),이용섭 ( Yong Sup Lee ) 경희대학교 동서약학연구소 2006 경희약대 논문집 Vol.34 No.1

큰까치수영의 구성성분

김진숙(Jin Sook Shin-Kim),김형자(Hyoung Ja Kim),박호군(Ho koon Park) 大韓藥學會 1993 약학회지 Vol.37 No.4

Four flavonoide glycosides and (-)-Epicatechin were isolated from the aqueous extracts of dried whole part of Lysimachia clethroides Duby(Primulaceae). They were 3-O-Methyl-quercetin-7-O-[alpha-L-rhamnopyranosyl (1-2) glucopyranoside], Quercetin-3-O-beta-D-glucopyranoside, Kaempferol-3-O-beta-D-glucopyranoside, Kaempferol-3-O-[alpha-L-rhamnopyranosyl (1-6)-beta-D-glucopyranoside] and (-)-Epicatechin. 3-O[alpha-L-rhamnopyranosy] (1-6)-beta-D-glucopyranoside] and (-)-Epicatechin. 3-O-Methyl-quercetin-7-O-[alpha-L-rhamnopyranosyl (1-2)-beta-D-glucopyranoside] and (-)-Epicatechin were first isolated from this plant. Their structures were elucidated by spectral analysis [1H-, 13C- NMR, 1H-1H-COSY, DEPT-analysis, HMQC(Heteronuclear multiple quantum coherence), FAB-MS].

자외선으로 유도된 Hs68 섬유아세포의 노화 반응에 대한 영실추출물의 억제 효능

박지은 ( Ji-eun Park ),김형자 ( Hyoung Ja Kim ),김수남 ( Su-nam Kim ),강승현 ( Seung Hyun Kang ),김연준 ( Youn Joon Kim ) 대한화장품학회 2015 대한화장품학회지 Vol.41 No.4

극심하고 지속적인 자외선에의 노출은 정상적인 피부구조를 파괴하는 다양한 피부 광노화 과정을 야기한다. 자외선은 인체 피부에서 세포외기질의 구성성분을 분해시키는 기질 분해효소인 matrix metalloproteinases (MMPs)의 발현을 활성화시키고, 콜라겐 합성은 감소시킴으로써 피부의 탄력과 구조적 치밀도를 약화시켜 궁극적으로 피부주름을 생성한다. 본 연구에서는 이러한 피부 광노화 현상을 완화시키는 소재로서 영실의 효능을 검증하고자 하였다. 먼저 인간 섬유아세포주인 Hs68을 이용하여 영실의 세포증식 촉진효능을 확인하였다. 여기에 더해 영실이 activator protein (AP)-1 전사인자의 억제를 통해 MMP의 발현을 감소시킴을 mRNA 및 단백질 수준에서 검증하였다. 또한, 진피층을 구성하는 타입 Ⅰ형 콜라겐과 표피-진피 경계부를 단단히 고정시키는 역할을 하는 타입 Ⅳ형 콜라겐 역시 영실에 의해 발현이 증가하며, 자외선에 의한 염증반응의 억제에도 영실이 효과적으로 작용하는 것을 확인할 수 있었다. 결론적으로 본 연구를 통해 영실이 자외선에 의한 피부노화와 주름 생성을 효과적으로 개선할 수 있는 가능성을 가짐으로써 항노화, 항염증 및 항주름 소재로서 화장품에 응용될 수 있을 것으로 기대된다. Acute and chronic ultraviolet (UV) irradiation triggers severe skin photoaging processes, which directly disrupt the normal three-dimensional integrity of skin. UV light stimulates the expression of matrix metalloproteinases (MMPs) which degrade constituents of extracellular matrix (ECM) proteins. These MMPs reduce collagen synthesis and decrease skin elasticity and integrity, resulting in wrinkle formation. In this study, we identified Rosa multiflora hip extract (RME) as an effective anti-photoaging ingredient. First, cell proliferation activity of RME was verified using Hs68 human dermal fibroblast cell line. RME downregulated MMPs expression through the inhibition of activator protein (AP)-1. In addition, type Ⅰ and Ⅳ collagen expressions were increased with RME treatment and UVB-induced inflammatory responses were also reduced after RME treatment. In conclusion, R. multiflora hip extract may effectively improve UVB-induced skin aging and wrinkle formation which may provide as an anti-aging, anti-wrinkle, and anti-inflammation ingredient in cosmetic industry.

전통 약용식물 및 각종 식물의 항암 효과에 대한 연구 (III)

현진원,임경화,성민숙,강삼식,백우현,배건우,조현,김형자,우은란,박호군,박재갑,양용만,Hyun, Jin-Won,Lim, Kyoung-Hwa,Sung, Min-Sook,Kang, Sam-Sik,Paik, Woo-Hyun,Bae, Kun-Woo,Cho, Hyun,Kim, Hyoung-Ja,Woo, Eun-Rhan,Park, Ho-Koon,Park, Jae-Gah 한국생약학회 1996 생약학회지 Vol.27 No.2

Antineoplastic activity against human gastric and colon carcinoma cell lines was tested in eighty-three species of Korean plants including Korean medicinal plants which have been frequently used in oriental herb prescriptions. The plant materials were extracted with methanol and the cytotoxic activity was tested using a calorimetric tetrazolium assay (MTT assay). Twenty-six plant extracts against gastric carcinoma cell line, eighteen extracts against colon carcinoma cell line and fourteen plant extracts against both carcinoma cell lines showed antineoplastic activity at the concentration of less than $100{\mu}g/ml$. The effective components from four species have been isolated and reported.

섬쑥부쟁이 에탄올 추출물의 잔틴산화효소 저해 효능 및 HPLC-UV를 이용한 유효성분의 함량 분석

강동현(Dong Hyeon Kang),한은혜(Eun Hye Han),진창배(Changbae Jin),김형자(Hyoung Ja Kim) 한국식품영양과학회 2016 한국식품영양과학회지 Vol.45 No.11

섬쑥부쟁이로부터 고요산혈증 개선에 도움을 주는 기능성 식품 개발을 위하여 최적의 에탄올 추출물 탐색과 high performance liquid chromatography-ultraviolet(HPLCUV) 분석방법에 의한 validation을 실시하였다. 지표성분으로 3,5-dicaffeoylquinic acid(3,5-DCQA)를 선정하여 표준화를 실시하였으며 검출법 확립을 위한 3,5-DCQA 정량분석은 Luna RP-18 칼럼(4.6×250 mm, 5 μm)을 이용하여 1% 초산용액과 메탄올을 전개용매로 사용하였다. 용출은 1.0 mL/min의 유속으로 기울기 용출(gradient elution) 방법을 이용하였으며, 320 nm 파장에서 검출한 피크 면적을 이용하여 검량곡선을 작성하여 분석하였다. 본 연구에서 확립한 분석법으로 특이성, 직선성, 정밀성, 정확성, 회수율을 검색하였다. 3,5-DCQA의 검량선으로부터 상관계수(R²)0.9999의 우수한 직선성과 intra-day와 inter-day 분석에서 90% 이상의 회수율과 5% 미만의 RSD를 나타내 정밀성과 정확성을 입증하였다. 검출한계는 2.68 μg/mL였고 정량한계는 8.11 μg/mL로 나타났다. 섬쑥부쟁이 에탄올 추출물(AGE)은 70과 80°C에서 30, 50, 70, 80% 에탄올로 3, 4,5, 6시간 동안 각각 추출하였으며, 지표물질의 검량곡선을 활용하여 각각의 AGE로부터 3,5-DCQA의 함량을 분석하였다. 본 시험법으로 분석한 3,5-DCQA의 함량은 70°C에서 추출한 70% AGE가 52.59±3.45 mg/Aster glehni 100g의 함량을 나타내 가장 우수하게 나타났다. 그러나 섬쑥부쟁이 추출물에 함유된 5-caffeoylquinic acid(5-CQA)의 함량 비교분석은 에탄올 함량이나 추출 시간에 따른 함량 변화가 미미하게 나타났다. 또한, 다양한 AGE에 대하여 XOD저해 효능을 검색하였을 때, 3,5-DCQA의 함량이 가장 높은 70°C에서 추출한 70% AGE에서 우수한 효능을 나타내 기능성 원료 표준화를 위한 적합한 분석법임이 검증되었다. 따라서 본 연구를 통하여 확립된 3,5-DCQA의 분석법은 섬쑥부쟁이 에탄올 추출물로부터 개별인정형 건강기능식품기능성 원료 개발을 위한 유용한 자료로 활용될 것으로 생각한다. This study aimed to establish an optimal extraction process and high performance liquid chromatography-ultraviolet (HPLC-UV) analytical method for determination of 3,5-dicaffeoylquinic acid (3,5-DCQA) as a part of materials standardization for the development of a xanthine oxidase inhibitor as a health functional food. The quantitative determination method of 3,5-DCQA as a marker compound was optimized by HPLC analysis using a Luna RP-18 column, and the correlation coefficient for the calibration curve showed good linearity of more than 0.9999 using a gradient eluent of water (1% acetic acid) and methanol as the mobile phase at a flow rate of 1.0 mL/min and a detection wavelength of 320 nm. The HPLC-UV method was applied successfully to quantification of the marker compound (3,5-DCQA) in Aster glehni extracts after validation of the method with linearity, accuracy, and precision. Ethanolic extracts of A. glehni (AGEs) were evaluated by reflux extraction at 70 and 80°C with 30, 50, 70, and 80% ethanol for 3, 4, 5, and 6 h, respectively. Among AGEs, 70% AGE at 70°C showed the highest content of 3,5-DCQA of 52.59±3.45 mg/100 g A. glehni. Furthermore, AGEs were analyzed for their inhibitory activities on uric acid production by the xanthine/xanthine oxidase system. The 70% AGE at 70°C showed the most potent inhibitory activity with IC50 values of 77.01±3.13∼89.96±3.08 μg/mL. The results suggest that standardization of 3,5-DCQA in AGEs using HPLC-UV analysis would be an acceptable method for the development of health functional foods.

손바닥선인장 추출물의 플라보노이드 구조 규명 및 HPLC-PDA를 이용한 지표성분의 함량 분석

박승배(Seungbae Park),강동현(Dong Hyeon Kang),진창배(Changbae Jin),김형자(Hyoung Ja Kim) 한국식품영양과학회 2017 한국식품영양과학회지 Vol.46 No.2

손바닥선인장으로부터 기능성식품 개발을 위하여 최적의 에탄올 추출물 탐색과 HPLC-PDA 분석방법에 의한 validation을 실시하였다. 지표성분으로 dihydrokaempferol (DHK)과 3-O-methylquercetin(3-MeQ)을 선정하여 표준화를 실시하였으며 검출법 확립을 위한 정량분석은 Luna RP-18 칼럼(4.6×250 mm, 5 μm)을 이용하여 1% 인산용액과 아세토니트릴을 전개 용매로 사용하였다. 용출은 1.0 mL/min의 유속으로 기울기 용출(gradient elution) 방법을 이용하였으며, 280 nm 파장에서는 DHK를, 360 nm 파장에서는 3-MeQ를 검출한 피크 면적을 이용하여 검량곡선을 작성하여 분석하였다. 본 연구에서 확립한 분석법으로 특이성, 직선성, 정밀성, 정확성, 회수율을 검색하였다. DHK의 검량선으로부터 상관계수(R2) 0.9998의 우수한 직선성과 intra-day와 inter-day 분석에서 97% 이상의 회수율과 3% 미만의 RSD를 나타내 정밀성과 정확성을 입증하였다. 3-MeQ의 검량선으로부터 상관계수(R2) 1의 우수한 직선성과 intra-day와 inter-day 분석에서 95% 이상의 회수율과 7% 미만의 RSD를 나타내 정밀성과 정확성을 입증하였다. DHK의 검출한계는 1.38 μg/mL, 정량한계는 4.18 μg/mL로 나타났으며, 3-MeQ의 검출한계는 3.49 μg/mL, 정량한계는 10.6 μg/mL로 나타났다. 손바닥선인장 에탄올 추출물 (OFSEs)은 70과 80°C에서 50, 70, 80% 에탄올로 3, 4, 5, 6시간 동안 각각 추출하였으며, 지표물질의 검량곡선을 활용하여 각각의 OFSEs로부터 두 종의 지표물질 함량을 분석하였다. 본 시험법으로 분석한 지표물질의 함량은 80°C에서 추출한 70% OFSE가 DHK 26.42±0.65 mg/OFS 100 g, 3-MeQ 3.88±0.29 mg/OFS 100 g의 함량을 나타내 가장 우수하게 나타났다. 다양한 OFSEs에 대하여 DPPH 자유 라디칼 소거효능과 쥐의 간 균질액을 이용한 지질과산화 저해 효능에 대한 항산화 효능은 지표물질의 함량이 가장 높은 70% OFSE에서 우수한 효능을 나타내 본 연구에서 확립한 원료 표준화를 위한 적합한 분석법임이 검증되었다. 따라서 본 연구를 통하여 HPLC-PDA를 이용한 손바닥선인장 에탄올 추출물의 DHK와 3-MeQ의 분석법은 개별인정형 건강기능식품 기능성 원료 개발을 위한 유용한 자료로 활용될 것으로 생각한다. This study aimed to establish an optimal extraction process and high-performance liquid chromatography (HPLC)-photodiode array (PDA) analytical method for determination of marker compounds, dihydrokaempferol (DHK) and 3-O-methylquercetin (3-MeQ), as a part of materials standardization for the development of health functional foods from stems of Opuntia ficus-indica var. saboten (OFS). The quantitative determination method of marker compounds was optimized by HPLC analysis, and the correlation coefficient for the calibration curve showed very good linearity. The HPLC-PDA method was applied successfully to quantification of marker compounds in OFS after validation of the method in terms of linearity, accuracy, and precision. Ethanolic extracts from stems of O. ficus-indica var. saboten (OFSEs) were evaluated by reflux extraction at 70 and 80°C with 50, 70, and 80% ethanol for 3, 4, 5, and 6 h. Among OFSEs, OFS70E at 80°C showed the highest contents of DHK and 3-MeQ of 26.42±0.65 and 3.88±0.29 mg/OFS 100 g, respectively. Furthermore, OFSEs were determined for their antioxidant activities by measuring 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and lipid peroxidation (LPO) inhibitory activities in rat liver homogenate. OFS70E at 70°C showed the most potent antioxidant activities with IC50 values of 1.19±0.11 and 0.89±0.09 mg/mL in the DPPH radical scavenging and LPO inhibitory assays, respectively. To identify active components of OFS, various chromatographic separation of OFS70E led to isolation of 11 flavonoids: dihydrokaempferol, dihydroquercetin, 3-O-methylquercetin, quercetin, isorhamnetin 3-O-glucoside, isorhamnetin 3-O-galactoside, narcissin, kaempferol 7-O-glucoside, quercetin 3-O-galactoside, isorhamnetin, and kaempferol 3-O-rutinoside. The results suggest that standardization of DHK in OFSEs using HPLC-PDA analysis would be an acceptable method for the development of health functional foods.

프로스타글란딘 유도체의 합성과 그의 생물학적 활성에 관한 연구 2. 위궤양과 위산분비에 대한 프로스타글란딘 유도체의 효과

조태순(Tai Soon Cho),이선미(Sun Mee Lee),함원훈(Won Hun Ham),이병무(Byung Mu Lee),김경례(Kyoung Rae Kim),지상철(Sang Cheol Chi),고준일(Jun Ill Ko),박인(In Park),오창영(Chang Young Oh),박호군(Ho Koon Park),김형자(Hyoung Ja Kim),이향우(H 한국응용약물학회 1995 Biomolecules & Therapeutics(구 응용약물학회지) Vol.3 No.1

The antiulcer effects of newly synthesized prostaglandin derivatives were investigated in various experimental ulcer models and on gastric secretion in rats. HK-3 and HK-4, PGE₂ derivatives, prevented the formation of acute gastric ulcer induced by ethanol or aspirin in pylorus-ligated rats. The ulcer formation was moderately inhibited by HK-1 and HK-2, PGF_(2α) derivatives, and aggravated by SK-1, SK-2 and SK-3, PGF_(2α) derivatives. HK-3 and HK-4 reduced the volume, acid output and pepsin output of gastric juice in pylorus-ligated rats. The gastric perfusion with physiologic saline(pH 6.0) showed relatively constant acid secretion and indomethacin increased the acid secretion. The acid secretion was markedly decreased by PGE₂ but PGF_(2α) caused little change. Prostaglandin derivatives, especially HK-3 and HK-4, significantly inhibited the acid secretion induced by indomethacin. The results show that, PGE₂ derivatives, HK-3 and HK-4, inhibit acid secretion and also have protective effects on gastric ulceration induced by ethanol or aspirin.

Saucernetin-7 and Saucernetin-8 Isolated from Saururus chinensis Inhibit the LPS-induced Production of Nitric Oxide and Prostaglandin E_(2) in Macrophage RAW264.7 Cells

Park, Hee-Juhn,Kim, Ryung-Gue,Seo, Bo-Rim,Ha, Joohun,Ahn, Byung-Tae,Bok, Song-Hae,Lee, Yong Sup,Kim, Hyoung Ja,Lee, Kyung-Tae WHO COLLABORATING CENTRE FOR TRADITIONAL MEDICINE 2003 東西醫學硏究所 論文集 Vol.2003 No.-

In the present study, we demonstrate that saucernetin-8(1) and saucernetin-7 (2), isolated from the underground parts of Sauiurus chinensis (Saururaceae), exerted potent inhibitory effects on LPS-induced nitric oxide (NO) and prostaglandin E_(2)(PGE_(2)) production in RAW 264.7 cells. Both compounds 1 and 2, known as dineolignans, also suppressed the expression of iNOS and COX-2 protein in a dose-dependent manner. Thus, this study suggests that compounds 1 and 2-mediated inhibition of iNOS and COX-2 expression may be one of the mechanisms responsible for the anti-inflammatory effects of the underground parts of Saururus chinensis.

In vivo Anti-Inflammatory and Antinociceptive Effects of Liriodendrin Isolated from the Stem Bark of Acanthopanax senticosus

Jung, Hyun-Ju,Park, Hee-Juhn,Kim, Ryung-Gue,Shin, Kyoung-Min,Ha, Joohun,Choi, Jong-Won,Kim, Hyoung Ja,Lee, Yong Sup,Lee, Kyung-Tae WHO COLLABORATING CENTRE FOR TRADITIONAL MEDICINE 2003 東西醫學硏究所 論文集 Vol.2003 No.-

In the present study, hriodendrin isolated by activity-guided fractionation from the ethyl acetate (EtOAc) extracts of the stern bark of Acanthopanax senticosus, was evaluated for anti-inflammatory and antinocieptive activities. Liriodendrin (5, 10 ㎎/㎏/day, p.o) significantly inhibited the increase of vascular permeability induced by acetic acid in mice and reduced an acute paw edema induced by carrageenan in rats. When he analgesic activity was measured by the acetic acid-induced writhing test and hot plate test, liriodendrin showed a dose-dependent inhibition in animal models. In addition, syringaresinol, the hydrolysate of liriodendrin, more potently inhibited the LPS-induced production of NO, PGE_(2) and TNF-α production of macrophages than liriodendrin. Consistent with these observations, the expression level of iNOS and COX-2 enzyme was decreased by syringaresinol in a concentration-dependent manner. These result suggest that the anti-inflammatory and antinociceptive effects of liriodendrin after oral administration were attributable to the in vivo transformation to syringaresinol, which may function as the active constituent.

Taraxinic Acid, a Hydrolysate of Sesquiterpene Lactone Glycoside from the Taraxacum coreanum NAKAI, Induces the Differentiation of Human Acute Promyelocytic Leukemia HL-60 Cells

CHOI, Jung-Hye,SHIN, Kyung-Min,KIM, Na-Young,HONG, Jung-Pyo,LEE, Yong Sup,KIM, Hyoung Ja,PARK, Hee-Juhn,LEE, Kyung-Tae WHO COLLABORATING CENTRE FOR TRADITIONAL MEDICINE 2002 東西醫學硏究所 論文集 Vol.2002 No.-

The present work was performed to elucidate the active moiety of a sesquiterpene lactone, taraxinic acid-1' O-β-D-glucopyranoside (1) from Taraxcum coreanum NAKAI on the cytotoxicity of vsrious cancer cells. Based on enzymatic hydrolysis and MTT assay, thr active moiety should be attributed to the aglycone taraxinic acid (la), rather than the glycoside (1). Taraxinic acid exhibited potent antiproliferative activity against human leukemia-derived HL-60. In addition, this compound was found to be a potent inducer of HL-60 cell differentiation as as-sessed by a nitroblue tetrazolium reduction test, esterase activity assay, phagocytic activity assay, morphology change, and expression of CDl4 and Cn66b surface antigens. Thrse results suggest that taraxinic acid induces the differentiation of human leukemia cells to monotyte/macrophaee lineage. Moreover, the expression level of c-myc was down-regulated during taraxinic acid-dependent HL-60 cell differentiation, whereas p21^CIPI and p27^KIPI were up-regulated. Taken together, our results suggest that taraxinic acid may have potential as a terapeutic agent in human leukemia.