• Natural Product Sciences
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• 제11권1호
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• pp.1-9
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• 2005

The chemical constituents from Hedyotis diffusa Willd and their pharmacological activities were summarized. It has been known data that this herb contains anthraquinones, terpenoids, steroids, flavonoids, organic acid, and polysaccharides. The studies of pharmacology have shown that Hedyotis diffusa Willd possess various levels of activities such as anticancer, anti-inflammatory, immunostimulatory, antioxidative, neuroprotective, and hepatoprotective activities.

• 한국식품영양과학회지
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• 제34권1호
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• pp.107-112
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• 2005

본 연구에서는 식중독 유발세균에 대한 항균활성이 우수한 천연 항균성 물질을 검색하기 위해 예로부터 민간과 한방에서 널리 이용되어 온 백화사설초를 각종 유기용매로 추출하여 식중독 유발세균에 대한 항균활성을 조사해 보았다. 백화사설초를 petroleum ether, chloroform, ethly acetate, methanol를 이용하여 실온에서 각각 용매 별로 계통 분획하고, 열수추출물을 얻은 후, 9종의 식중독 유발세균(Staphylococcus aureus, nacillus cereus, Salmonella enteritidis, Shigella nexneri, Escherichia coli, Salmonella Typhimurium, Shigella dysenteriae, Pseudomonas aeruginosa, Shigella sonnei)에 대하여 항균효과를 조사하였다 백화사설초 추출물의 농도별 항균 활성 검색에서는 백화사설초의 methanol추출물이 가장 큰 항균 효과를 보였으며 S. aureus와 S. flexneri가 가장 민감하게 반응하는 균주였다. 백화사설초의 methanol추출물과 산두근의 ethyl acetate추출물을 혼합하여 항균력을 측정해 본 결과 두 추출물을 섞어 첨가했을 경우가 단독으로 사용했을 시보다 상승효과를 나타내었다. 또한 백화사설초의 methanol추출물이 식중독 유발세균의 성장에 미치는 효과를 검정하기 위해 S. aureus 및 S. Typhimurium의 배양액에 백화사설초의 methanol 추출물을 각각 4,000 ppm농도로 첨가했을 시, S. aureus의 생육이 36시간 이상까지 억제됨을 관찰할 수 있었고, S. Typhimurium의 생육도 24시간까지 지연시킬 수 있었다. 이상의 결과 백화사설초의 methanol추출물은 S. aureus와 S. Typhimurium의 성장을 효과적으로 억제시킴을 알 수 있었다.

• 대한한방종양학회지
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• 제6권1호
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• pp.99-111
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• 2000

Objectives: Hedyotis diffusa is used to treat cancer in traditional Korea Medicine. So this study was carried out to examine the expression of cell cycle related genes in HL-60 cells undergoing apoptosis by the methanol extract of Hedyotis diffusa. Methods: 1. HL-60 cells were treated with various concentrations (from 200 to $50{\mu}g/ml$)of methanol extract and H20 extract ($200{\mu}g/ml$) of hedyotis diffusa. After 48 h later, the cells were tested for viability by MTT assay. 2. The HL-60 cells were treated with $200{\mu}g/ml$ of methanol extract for the indicated periods. The whole cell lysates were prepared and analyzed by western blotting using anti-p53 antibody. 3. The nuclear extract were prepared and analyzed by western blotting using anti-p21 antibody, anti-p27 antibody, anti-cyclin A antibody, anti-cyclin E antibody and anti-CDK2 antibody. Results: 1. The methanol extract of Hedyotis diffusa induced the death of HL-60 cells in a dose dependent manner. 2. The methanol extract of Hedyotis diffusa makedly decreased the level of p21/Cipl and cyclin A in a time dependent manner. 3. The methanol extract of Hedyotis diffusa markedly increased the level of p27/Kipl and cyclin E in a time dependent manner. 4. The methanol extract of Hedyotis diffusa markedly did not affect the level of CDK2. Conclusions: These results provide evidence that expression of cell cycle related genes in HL-60 cells undergoing apoptosis by the methanol extract of Hedyotis diffusa mainly results from decreased level of p21/Cipl and increased level of p27/Kipl of the cell cycle related genes.

• 대한한방종양학회지
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• 제6권1호
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• pp.81-97
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• 2000

Objectives: This experimental study was carried out to evaluate the effects of aqueous and methanol extracts of Hedyotis diffusa which has long been used for cancer treatment in oriental medicines on the induction of apoptotic cell death in human lymphoid leukemia cell line, HL-60. Methods: Cells were treated with various concentrations (200 to $0.4{\mu}g$) and periods (6 to 30 hr) of $H_2O$ and methanol extracts of Hedyotis diffusa. Then, cells were tested for viability by MTT assay. Cells wrere treated with $200{\mu}g/ml$ of methanol extract fork various periods. Genomic DNA was isolated, separated, on 1.5% agarose gels, stained with ethidium bromide and visualized under UV light. Cells were treated with $200{\mu}g/ml$ of each extract for 16 hr. Then, cells were treated with Hoechst dye 33342 and observed by fluorescence microscopy. Cells were treated with various doses of each for 12 hr and $100{\mu}g/ml$ of methanol extract for various periods. Lysate from the cells used to measure the activity of Caspase-1 and-3 proteases by using fluorogenic peptide substrates including acetyl-YVAD-AMC and acetyl-DEVD-AMC, respectively. Cells were treated with $200{\mu}g/ml$ of each extract for various periods. Cell lysates were immunoprecipated with anti-JNKl antibodies. The immune complex was reacted with $32^p-ATP$ and c-Jun as a substrate. The phosphotransferase activity of JNKI was measured by using PhosphoImage analyzer (Fuji Co., Japan). Nuclear extracts were isolated and incubated with oligonucleotide probe of $NF-{\kappa}B$. Transcriptional activation of ${\kappa}B$ was measured by using EMSA and visualized by PhosphoImage analyzer (Fuji Co, Japan). Cell lysates were prepared and analyzed by Western blotting with anti-Bc12 antibodies and anti-Bax antibodies. Cells were pretreated with various doses of methanol extract for 2 hr. Then, the extract was removed by centrifugation. Cells were resuspended with RPMI-1640 media containing 0.3% agarose, 10% FBS, overlayred onto bottom layer agarose and incubated at $CO_2$ incubator for 6 days. The number of colony was counted under light microscopy ($\time100$). Results: The death of HL-60 cells was markedly induced by the addition of methanol extract of Hedyotis diffusa in a dose and time-dependent manners. The apoptotic characteristic ladder pattern of DNA strand break was observed in death of HL-60 cells. In addition, it was shown nucleus chromatin condensation and fragmentation under Hoechst staining. Therefore, Hedyotis diffusa extract-induced death of HL-60 cells is mediated by apoptotic signaling processes. The activity of Caspase 3-like proteases remained in a basal level in HL-60 cells treated with aqueous extract of Hedyotis diffusa. However, it was markedly increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. In addition, the phosphotransferase activity of JNKl was increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. Furthermore, the activation of transcriptional activator, $NF-{\kappa}B$ was markedly induced by methanol extract of Hedyotis diffusa. Anti-apoptotic Bc12 was cleaved into 23Kda fragment by treatment of methanol extract of Hedyotis diffusa. However, expression of proapoptotic Bax protein was increased by treatment of methanol extract of Hedyotis diffusa in a time-dependent manner. Furthermore, methanol extract markedly inhibited the colony forming efficiency of HL-60 cells in semisolid agar culture. Conclusions: Above results suggest that methanol extract of Hedyotis diffusa induces the apoptotic death of human leukemic HL-60 cells via activations of Caspase-3 proteases, JNKI, transcriptional activator $NF-{\kappa}B$, In addition, our results also suggest that methanol extract of Hedyotis diffusa reduces the malignant potential of HL-60 cells via down regulation of colony forming effciency through cleavage of Bc12 as well as induction of Bax.

• 대한한의학방제학회지
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• 제8권1호
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• pp.299-318
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• 2000

In order to study on SOD like activity and Electric donor ability of Hedyotis diffusa $W_{ILLD}$, testing antibacterial effect on E. coli O157 which causes infectious inflammatory disease; measunng inhibitory effect on tyrosinase which stimulates melanin formation; and measuring the influence of it to SOD like activity, DPPH and TBARS which are related to ageing and carcinogenesis. The results of these experiments, are as follows. 1. In the antibacterial experiment with the extract of Hedyotis diffusa $W_{ILLD}$ on E. coli O157, there was no change in proliferation from the beginning of culture to two hours after, but proliferation inhibiting effect on E. coli O157 was detected from three hours after the beginning of culture. 2. Tyrosinase inhibitory effect was measured as $0.39{\pm}0.026%$. Compared with the control group, the effect was very slight. 3. The SOD like activity of the extract of Hedyotis diffusa $W_{ILLD}$. was measured as 21.33${\pm}$ 4.264%. Compared with the extracts of several other herbs, it was much more significant. 4. The DPPH of the extract of Hedyotis diffusa $W_{ILLD}$. was $53.3{\pm}0.91%$ when 0.02g was used and $83.5{\pm}1.82% $ when 0.05g was used. The result when 0.05g was used was more significant than 0.02g was used. 5. The TBARS of the extract of Hedyotis diffusa $W_{ILLD}$. was measured as 0.724 ${\pm}$0.O04MDAppm. Compared with the extracts of several other herbs, the result was more significant. From these results, we found that Hedyotis diffusa $W_{ILLD}$. can be used in therapy of dysentery with bloody stool and fever which is caused by infection with E. coli O157 and that it can be also used effectively as age resister or anticarcinogen.

• The Korean Journal of Ecology
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• 제17권4호
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• pp.523-531
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• 1994

Optimal conditions for the seed germination and growth of Hedyotis diffusa were studied. As photoperiod was increased from 12 hr to 24hr, the germination rate of Hedyotis diffusa was gradually increased. The photoperiod and temperature inflenced on the fermination synergistically. After the growth of 20 weeks under the natural condition (June~Oct.), the length of H. diffusa was $38.9{\pm}4.2cm$ (15.5~52.5cm), and total dry weight per $3.3m^2$ was $316.7{\pm}10.3g$. It is considered that H. diffusa could be cultivated in a part of inland. The anticancer effect of H. diffusa extract was examined. F-344 rats aged 6 weeks were divided into 3 groups and were given an I.P. of diethylnitrosamine at 200mg/kg body weight as a promoter, initially. And in two weeks after the beginnign of the experiment, group 1 was supplied iwth feed containing 0.02% 2-AAG as a promoter for 6 weeks. Group 2 was supplied with feed containing extracts of H.diffusa (0.02%) for two weeks. Group3 was supplied with only basal diet. All rats were sacrificed for partial hepatectomu, and the antipromoting effect was examined by the number 문 area per $cm^2$ of foci in river. In group 1, the number of hyperplastic nodule was $18.5{\pm}7.7$, but in group2, it was drastically reduced to $10.3{\pm}1.8$ rather thn those of group1. The total area of nodules $(mm^2)$ /whole liver $(cm^2)$ of group 1 and group2 were $19.2{\pm}7.7$ and $5.0{\pm}3.2$, respectively. These results indicate that extract of H.diffusa act as an anticancer agent at statistically significant level (p<0.001).

• Natural Product Sciences
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• 제9권1호
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• pp.7-9
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• 2003

The antioxidative constituents isolated from Hedyotis diffusa were identified as quercetin 3-O-${\beta}$-rutinoside (1) and quercetin 3-O-${\beta}$-glucoside (2). We also isolated asperuloside (3) from this plant. Identification was done based on spectroscopic analysis. Quercetin 3-O-${\beta}$-rutinoside was the stronger antioxidant than quercetin 3-O-${\beta}$-glycoside while asperuloside was inactive.

• 대한한방종양학회지
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• 제6권1호
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• pp.67-79
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• 2000

Objective : Hedyotis diffusa has been used as an anticancer agent for several decades in oriental medicine. We test whether the methanol extract of the herb affects transcriptional activation factors including $NF-{\kappa}B$ and AP-1. Methods : 1. HL-60 cells were treated with various concentrations(from 200 to $50{\mu}g/ml$) of methanol extract and $H_2O$ extract($200{\mu}g/ml$)of hedyotis diffusa, After 48h later, the cells were tested for viability by MTT assay. 2. The HL-60 cells were treated with $200{\mu}g/ml$ of methanol extract for the indicated periods. First. Nuclear extracts were isolated and incubated with oligonucleotide probe of $NF-{\kappa}B$ and AP-1. Second. Nuclear extracts were isolated and reacted with p50, p65. c-rel pan-Jun, c-Jun, JunB. JunD antibody on ice for 30min. Finally The cell lysates were prepared and analyzed by western blotting using anti-Fas, anti-FasL and anti-p53 antibody. Results : 1. The methanol extract decreases the viability of human lymphoid origin leukemia HL-60 cells in a dose-dependent manner. 2. $NF-{\kappa}B$ is rapidly activated by the addition of the methanol extract, reaches a peak at 30min and gradually returns to resting level. We confirm that $NF-{\kappa}B$ is a heterodimer mainly composed of p65 subunit with c-Rel. 3. Transcriptional activation of AP-1 is detected at 30min and reaches a maximum at 1hr after stimulation of the cells with the methanol extract. AP-1 is mainly composed with Jur-D and partially Jug-B proteins. 4. the methanol extract of Hedyotis diffusa induces the expression of Fas, Fas ligand and p53 proteins of HL-60 cells in a time dependent fashion. Conclusions : These results suggest that the methanol extract of Hedyotis diffusa exerts anticancer effects to induce the death of human leukomic HL-60 cells via activation of trascriptional factors such as $NF-{\kappa}B$ and AP-1, increase in expression of Fas mediated signalling proteins, and induction of tumor suppressor gene. p53.

• 대한약학회:학술대회논문집
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• 대한약학회 2000년도 춘계총회 및 학술대회
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• pp.192.2-192.2
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• 2000

• 생약학회지
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• 제34권4호통권135호
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• pp.288-292
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• 2003

We compared the antitumor activity between hollow fiber assay and xenographic animal assay on thirty herbal plants. It is evaluated that the antitumor activity of above 30% is regarded as 'positive response', and its of below 30% is regarded as 'negative response'. The two herbal plants extracts (Ulmus davidiana Hedyotis diffusa) among thirty herbal plants show to be positive in xenographic animal assay and they were also correctly identified as positive by the hollow fiber assay. The correlation of the hollow fiber assay data with xenographic animal assay would suggest that hollow fiber assay presents a potentially unique tool to develop the herbal medicine for cancer.