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박혜란,함연호,이성태,백상기,조성기,Park, Hae-Ran,Ham, Yeon-Ho,Yee, Sung-Tae,Paik, Sang-Gi,Jo, Sung-Kee 대한면역학회 2001 Immune Network Vol.1 No.2
Background : Paeonia japonica Miyabe is a medicinal plant which has been widely used as a component of blood-building decoctions (Chinese medicinal concept : Bu-Xie). The immunopharmacological characteristics of the extract of Paeonia japonica (PJ) were investigated. Methods : The effects of fractions of PJ extract on lymphocyte proliferation were measured by $H^3$-thymidine incorporation assay. The proliferated lymphocyte subsets were analyzed in flow cytometry. The subset cell populations of spleen cells were separated by magnetic cell separation system, and their proliferation by the extract were investigated. The effect of the extract on antibody production was determined in mice challenged with sheep red blood cells (SRBC) using hemolytic plaque forming cell assay. Results : Spleen cells were proliferated by water extract of PJ. Polysaccharide fraction (PJ-P) of the extract was most active in the proliferation. It was found in flow cytometry that the lymphocyte subset proliferated by PJ-P was B cell population. Among the separated subset cell populations, T cell-depleted cell population and macrophage-depleted cell population were most proliferated by PJ-P. However, positively selected populations of B cells and T cells were not proliferated by PJ-P. These results indicate that B cell proliferation by PJ-P may require the assistance of macrophages or T cells. These results suggest that firstly PJ-P may stimulate macrophages or T cells, and then B cells are activated. The number of antibody-secreting cells was increased by administration of PJ-P in mice immunized with SRBC as a T-dependent antigen. Conclusion : These results suggest that macrophages and accessory cells are directly activated by PJ-P and then helper T cells and B cells are indirectly activated. As the results, immune responses might be coordinately improved. In conclusion, PJ-P, a polysaccharide of P. japonica, may be a characteristic immunostimulator, which is analogous to polysaccharides such as lentinan, PSK and ginsan.
편백나무(Chamaecyparis obtusa Endlicher) 잎의 추출성분
이상극,김진규,함연호,배영수,Lee, Sang-Keug,Kim, Jin-Kyu,Ham, Yeon-Ho,Bae, Young-Soo 강원대학교 산림과학연구소 2001 Journal of Forest Science Vol.18 No.-
편백나무 잎을 채취하여 건조시킨 후 분말로 제조하여 아세톤-물(7:3, v/v)로 추출하고 헥산, 메틸렌클로라이드, 에틸아세테이트, 그리고 수용성으로 분획하여 동결건조 시켰다. 그 중에서 에틸아세테이용성 분획을 Sephadex LH-20으로 충진한 칼럼에서 메탄올과 에탄올-헥산 혼합액을 용매로 사용하여 칼럼크로마토그래피를 실시하였다. 단리된 화합물들은 박층크로마토그래피(TLC)로 확인한 후 NMR스펙트럼을 사용하여 정확한 구조규명을 하였고 FAB-MS스펙트럼으로 분자량을 측정하였다. 주로 많은 양의 taxifolin-3-O-${\beta}$-D-xylopyranoside와 (+)-catechin이 포함되어 있었으며 소량의 quercetin-3-O-${\alpha}$-L-rhamnopyranoside도 함께 단리 되었다. 2kg of the dried leaves of chamaecyparis obtusa Endlicher were ground, extracted with acetone-$H_2O$(7:3, v/v), concentrated, and fractionated with a series of hexane, $CH_2Cl_2$, EtOAc and water on a separatory funnel. Each fraction was freeze dried to give dark-brown powder and a EtOAc soluble portion of the powder was chromatographed on a Sephadex LH-20 column using a series of aqueous methanol and ethanol-hexane mixture as eluents. Spectrometric analyses such as NMR and FAB-MS including TLC were performed to characterize the structures of the isolated compounds. The leaves of chamaecyparis obtusa Endlicher contained a large amount of taxifolin-3-O-${\beta}$-D-xylopyranoside and (+)-catechin in addition to a small amount of quercetin-3-O-${\alpha}$-L-rhamnopyranoside.
감마선조사 감초, 진피(陳皮) 및 시호 열수 추출물의 in vitro 유전독성학적 안전성 평가
조성기(Sung-Kee Jo),함연호(Yeon-Ho Ham),박혜란(Hae-Ran Park),오헌(Heon Oh),변명우(Myung-Woo Byun) 한국식품영양과학회 2001 한국식품영양과학회지 Vol.30 No.6
생약재의 기능성 식품 및 대체의약 원료로의 이용 증대에 따라 위생적 저장ㆍ유통을 위한 감마선조사 기술의 이용 가능성을 검토할 목적의 일환으로 실제 이용선량의 최고 선량인 10 kGy의 감마선 조사 생약재 3종의 유전독성학적 안전성을 평가하고자 하였다. 공시 재료는 감마선 조사된 생약재료 감초, 진피 및 시호로 하였다. 시험은 Salmonella typhimurium 균주를 이용한 유전자 복귀돌연변이 시험(Ames test)과 배양된 Chinese hamster ovary(CHO) 세포를 이용한 in vitro 소핵유발 시험으로 시행하였다. 시료는 오염유기체 완전 구제 선량인 10 kGy의 감마선으로 조사된 감초, 진피 및 시호의 열수 추출물이었으며, 시료의 농도는 복귀돌연변이 시험의 경우 5 mg/plate로, 소핵유발 시험의 경우 50%의 세포증식 억제를 나타내는 농도를 최고 농도로 하였다. 시험은 대사 활성화시키지 않은 경우와 S9 mix 첨가로 대사 활성화시킨 경우로 나누어 시행하였다. 복귀돌연변이 시험 결과 각 시료에 의한 복귀변이 집락수의 증가를 인정할 수 없었으며, 각 용량단계에서 감마선 비조사군과 조사군간의 차이도 볼 수 없었으므로 음성으로 판정하였다. 소핵유발 시험에서 cytokinesisblocked binucleated(CB) cells 내에 생성된 소핵을 계수한 결과, 음성 대조군의 경우 소핵 출현빈도가 20~30/1,000 CB cells(2~3%) 정도였으며, 비조사 시료군과 감마선조사 시료군의 각 용량단계에서 모두 2~4%의 소핵 출현빈도를 보여 시료에 의한 소핵 출현빈도의 증가를 인정할 수 없었다. 따라서 감마선이 조사된 각 시료가 직접변이원이나 간접변이원으로 작용하지 않으며, 세포분열 중에 유전학적으로 독성을 나타내지 않음을 확인할 수 있었다. 이 결과로 보아 생체내 유전독성시험, 만성독성시험 및 생식독성시험 등이 추가된다면 감마선 조사 생약재의 안전성을 명확히 밝힐 수 있을 것으로 사료된다. The γ-irradiated medicinal herbs were examined the genotoxicological safety to consider the possibility of application of the irradiation technology for hygienic purpose. The three medicinal herbs -Glycyrrhizae Radix, Aurantii nobilis Pericarpium and Bupleuri Radix- were irradiated with γ-rays at the practical dosage of 10 kGy. The hot water extracts of the irradiated herbs were examined in two short-term in vitro tests: (1) Ames test in Salmonella typhimurium TA98 and TA100, (2) Micronucleus test in cultured Chinese hamster ovary(CHO) cells. In the Salmonella reversion assays both with and without metabolic activation, the number of revertant colonies was not increased with each extract from the irradiated herbs, compared with negative controls. No significant difference in formation of the colonies was observed between non-irradiated and 10 kGy-irradiated herbs. These results indicated that no mutagenicity of the irradiated herbs was detected. In the micronucleus tests in cultured CHO cells, the incidences of micronucleus were not increased with irradiated herbs, and no significant difference in the incidences was observed between non-irradiated and irradiated herbs. These results indicated that no cytogenetic toxicity of the irradiated herbs was detected. The results of the two in vitro tests suggest that the irradiated herbs do not show mutagenic effects and cytogenetic toxicity. Further tests of in vivo genotoxicity and chronic toxicity are needed to determine the safety of the herbs irradiated with γ-rays at practical doses.