간암 세포주 HepG2에 대한 맥문동탕(麥門冬湯)추출물의 항암 및 항전이 효능

전명숙 ( Myeong Sook Cheon ),천진미 ( Jin Mi Chun ),윤태숙 ( Tae Sook Yoon ),이아영 ( A Yeong Lee ),문병철 ( Byeong Cheol Moon ),추병길 ( Byung Kil Choo ),김성환 ( Seong Hwan Kim ),김호경 ( Ho Kyoung Kim ) 대한본초학회 2009 大韓本草學會誌 Vol.24 No.3

Objectives: Maekmoondong-tang (MMDT), a Korean herbal medicine, has been used to treat severe dry cough in patients with bronchitis and pharyngitis. MMDT has been reported to have anti-inflammatory, anti-allergic, immunomodulatory, secretory-modulating, and metabolic regulatory actions. However, there are no evidence in regard to the effects of MMDT on carcinogenesis and metastasis. Here, we investigated the effects of 70% ethanol extract of MMDT on cell viability, apoptosis, and motility in human hepatocarcinoma HepG2 cells. Methods: Cell viability was measured using the CCK-8 assay, and the apoptosis induction was evaluated by caspase-3 activity. To detect apoptotic features, the cells treated with MMDT were stained with 4`-6-diamidino-2-phenylindole (DAPI). Cell motility was examined by Boyden chamber assay and Real-time Cell Index of Migration assay. Gelatin zymography also performed to measure matrix metalloproteinase (MMP)-2/9 activity. Results: We found that MMDT significantly inhibited cell proliferation and increased caspase-3 activity in a dose-dependent manner in HepG2 cells. Apoptotic features such as chromatin condensation and apoptotic bodies were observed in MMDT-treated cells by DAPI staining. MMDT also suppressed PMA-induced cell motility and activities of MMP-2/9. Conclusions: Our results exhibited that MMDT possess the anti-carcinogenetic and anti-metastatic activities via caspase-3 activation and down-regulation of cell motility and invasion in HepG2 cells. Therefore, these findings suggest that MMDT could be potentially applied to the prevention and treatment of cancer.

4종 대황(大黃)류 추출물의 항혈전 효능 비교

양원경 ( Won Kyung Yang ),김호경 ( Ho Kyoung Kim ),성윤영 ( Yoon Young Sung ),전명숙 ( Myeong Sook Cheon ),윤태숙 ( Tae Sook Yoon ) 대한본초학회 2010 大韓本草學會誌 Vol.25 No.4

Objective: The purpose of this study is to compare anti-thrombotic activities of 4 Rhubarb species. Methods: Rhubarb has been used as medical and edible resources worldwide for the therapy of cancer, constipation and inflammation, etc. To compare the anti-thrombotic activities among the species within rhubarb, we investigated and compared the inhibitory properties of water extracts from R. palmatum(RPE), R. officinale(ROE), R. nobile(RNE), and R. franzenbachii (RFE) on the fibrinolysis, blood coagulation and platelet aggregation. APTT and PT, blood coagulation time, were measured by apparatus of auto blood coagulation analysis that called ACL-7000. The measurement of fibrinolysis was measured and compares with the width of clear zone from melted fibrin plate. The platelet aggregation was measured and compared with inhibition rate of aggregation. Results: We found out the best species from rhubarb showing the significant fibrinolytic, anti-coagulant and anti-platelet aggregation activity. Our results show that Rheum nobile (RNE) has the most effective anti-thrombotic activity among 4 tested rhubarb. Conclusions: This study, therefore, RNE will be able to be used the development of drug which is thrombosis treatment.

강활과 방풍의 항염증 상승작용에 관한 연구

이도연 ( Do Yeon Lee ),최고야 ( Go Ya Choi ),윤태숙 ( Tae Sook Yoon ),전명숙 ( Myeong Sook Cheon ),김승주 ( Seung Ju Kim ),야스카와켄 ( Ken Yasukawa ),유소연 ( So Yeon Yu ),김호경 ( Ho Kyoung Kim ) 대한본초학회 2008 大韓本草學會誌 Vol.23 No.4

Objectives: Notopterygium incisum (N. incisum) and Saposhnikovia divaricata (S. divaricata) have been clinically used in traditional oriental medicine for treatment of inflammatory diseases. Also, a herbal mixture prepared with N. incisum and S. divaricata has been strongly linked to the anti-inflammatory effect. In this study, we evaluate the synergistic anti-inflammatory effect of N. incisum and S. divaricata. Methods: For evaluating the anti-inflammatory activity of a herbal mixture of N. in incisum and S. divaricata in vivo, we measured the changed ear thickness in 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-induced mouse ear edema model after topical application of herbal mixture. In addition, the levels of markers for inflammation, such as tumore necrosis factor (TNF)-α, interleukin (IL)-1β, and nitric oxide (NO), were determined by ELISA assay in lipopolysaccharide-stimulated Raw 264.7 cells. Results: We reported that water extracts of N. incisum and S. divaricata combination significantly inhibited the mouse ear edema induced by TPA. Moreover, the water extracts of N. incisum and S. divaricata combination exhibited synergistic effects in down-regulating IL-1β level, but not TNF-α and NO. Conclusions: These results suggest that combined treatment of N. incisum and S. divaricata, based on seven methods in prescription compatibility, has a synergistic effect in down-regulating inflammatory response both in vivo and in vitro models. Especially, it seems that IL-1β is a one of main target of the mixture of N. incisum and S. divaricata on anti-inflammatory activity.

rDNA-ITS 염기서열 분석을 통한 시호 종 감별용 유전자 마커 개발 및 유연관계 분석

문병철 ( Byeong Cheol Moon ),추병길 ( Byeong Kil Choo ),지윤의 ( Yun I Ji ),윤태숙 ( Tae Sook Yoon ),이아영 ( A Young Lee ),전명숙 ( Myeong Sook Cheon ),김보배 ( Bo Bae Kim ),김호경 ( Ho Kyoung Kim ) 대한본초학회 2009 大韓本草學會誌 Vol.24 No.3

Objectives: Bupleuri Radix (Siho) is prescribed as the root of different Bupleurum species on the pharmarcopoeia in Korea and China. Moreover, other species and varieties of the genus Bupleurum have been also distributed on the herbal market as Bupleuri Radix. However, due to the morphological similarity and frequent occurrence of intermediate forms, the correct identification of this radix is very difficult. To develop a reliable method for correct identification and improving the quality standards of official Bupleuri Radix, we analyzed sequences of the ribosomal RNA gene and internal transcribed spacer (rDNA-ITS) region. Methods: PCR amplification of rDNA-ITS region was performed using ITS1 and ITS4 primer from 6 Bupleurum species and 1 variety, B. falcatum L. (Siho), an improved breed of B. falcatum L. (Samdo-Siho), B. chinense DC. (Buk-Siho), B. scorzonerifolium Willd. (Nam-Siho), B. longiadiatum Turcz. (Gae-Siho), B. euphorbiodes Nakai (Deungdae-Siho) and B. latissimum Nakai (Seom-Siho), and nucleotide sequence was determined after sub-cloning into the pGEM-Teasy vector. Authentic marker nucleotides were estimated by the analysis of ClastalW using entire rDNA-ITS sequence of three samples per species. Results: In comparative analysis of the rDNA-ITS sequences, we found specific nucleotides to distinguish Korean (B. falcatum L. and its variety) and Chinese official species (B. chinense DC. and B. scorzonerifolium Willd.) from others at positions 411 and 447, and positions 89, 101, 415 and 599, respectively. Futhermore, we also found nucleotide indels (insertion and/or deletion) and substitutions to identify each of different Bupleurum species, 2 positions for B. falcatum L. and its variety, 6 positions for B. chinense DC., 49 positions for B. scorzonerifolium Willd., 8 positions for B. euphorbioides Nakai, 7 positions for B. longiradiatum Nakai and 9 positions for B. latissimum Nakai. These sequence differences at corresponding positions are avaliable nucleotide markers to determine the botanical origins of Bupleuri Radix. Moreover, we confirmed the phylogenetic relationship of B. latissimum Nakai, a Korean endemic speices, among Bupleurum species based on the rDNA-ITS sequence. Conclusions: These marker nucleotides would be useful to identify the official herbal medicines by the providing of definitive information that can identify each plant species and distinguish it from unauthentic adulterant Bupleurum species.

RAPD 분석을 통한 대황(大黃)과 종대황(種大黃) 감별용 SCAR 유전자 마커 개발

문병철 ( Byeong Cheol Moon ),이영미 ( Young Mi Lee ),천진미 ( Jin Mi Chun ),이아영 ( A Young Lee ),윤태숙 ( Taesook Yoon ),전명숙 ( Myeong Sook Cheon ),추병길 ( Byung Kil Choo ),김호경 ( Ho Kyoung Kim ) 대한본초학회 2009 大韓本草學會誌 Vol.24 No.4

Objectives: Due to the morphological similarity and frequent occurrence of intermediate forms as well as morphological variations of aerial part, the correct identification between Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma is very difficult. To develop a reliable method for correct identification and improving the quality standards of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma, we analyzed RAPD and developed SCAR marker. Methods: To amplify target DNA at the genomic level, 32 Operon 10-mer random primers were applied with four Rheum species, R. officinale, R. palmatum, R. tanguticum and R. undulatum. The nucleotide sequences were determined and species-specific primers were prepared depending on the species-specific RAPD amplicons after subcloned into the pGEM-Teasy vector. To develop the SCAR markers, species-specific PCR amplification and mutiplex-PCR were carried out using the single species-specific primer pairs and combinations of them, respectively. Results: We used RAPD analysis of four Rheum plant species to obtain several species-specific RAPD amplicons. From nucleotide sequences of these RAPD amplicons, we developed two SCAR markers that amplified 314 bp and 390 bp DNA fragments in only R. undulatum but not in R. officinale, R. palmatum, R. tanguticum and R. undulatum, for distinguishing Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma. Furthermore, we established SCAR markers for the simultaneous discrimination of the three species within a single reaction by using multiplex-PCR. Conclusions: These genetic markers can be used for the efficient discrimination of plants species and commercial herbal medicines between Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma, to ultimately prevent indiscriminate distribution and prescription of these herbal medicines.

강활(羌活)과 위령선(威靈仙)의 항염증 상승작용에 관한 연구

김승주 ( Seung Ju Kim ),김호경 ( Ho Kyoung Kim ),천진미 ( Jin Mi Chun ),양원경 ( Won Kyung Yang ),전명숙 ( Myeong Sook Cheon ),성윤영 ( Yoon Young Sung ),박준언 ( Jun Yeon Park ) 대한본초학회 2010 大韓本草學會誌 Vol.25 No.4

Objective: Oriental medicines have been combined oriental medical theory which based on the seven modes of emotions. Notopterygium incisum (N. incisum) and Clematis manshurica (C. manshurica) have been used as an anti-rheumatic and analgesic medicine for the treatment of rheumatism, headache, cold, etc. In this study, we evaluate the synergistic anti-inflammatory effect of N. incisum and C. manshurica. Method: To evaluate the synergistic anti-inflammatory effect of a herbal mixture N. incisum and C. manshurica, we examined the changed ear thickness in 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-induced mouse ear edema model after topical application of herbal mixture. In addition, the levels of markers for inflammation, such as tumor necrosis factor (TNF)-a, interleukin (IL)-1b, prostaglandin G2 (PGE2), and nitric oxide (NO) were measured by ELISA assay and Griess reagent in lipopolysaccharide-stimulated Raw 264.7 cells. Results: Our results showed that aqueous extracts of N. incisum and C. manshurica combination significantly inhibited the mouse ear edema induced by TPA. Moreover, the aqueous extracts of N. incisum and C. manshurica combination exhibited synergistic effects in down-regulating TNF-a, IL-1b, PGE2 level, but not NO. Conclusions : This study suggested that combined treatment of N. incisum and C. manshurica, based on seven methods in prescription compatibility, has a synergistic effect in down-regulating inflammatory response both in vitro and in vivo models.