• Journal of Plant Biotechnology
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• 제38권4호
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• pp.301-307
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• 2011

The medicinal herb Oldenlandia diffusa is known as a folk medicine for the treatment of hepatitis, sore throat, appendicitis, malignant tumors and urethral infection in Southern China and Korea. Another species O. corymbosa, is also used for the therapy of the similar conditions, however, only O. diffusa is referred to the medicinal herb by Chinese Pharmacopoeia. Due to their similar morphology, O. diffusa and O. corymbosa are often misidentified. To easily identify O. diffusa from O. corymbosa, the phylogenetic utility of nuclear ribosomal DNA (nrDNA) internal transcribed spacers (ITS) were investigated among different O. diffusa and O. corymbosa populations in Korea. The nrDNA ITS sequence of O. diffusa contained 791 bp, with GenBank accession number of JF837601-JF837602. The nrDNA ITS sequence of O. corymbosa was 785-786 bp, with GenBank accession number of JF837603-JF837611. The results showed that there are some certain divergences in the ITS region sequence between both species, even among different populations of the same species. Particularly, O. corymbosa ST-4 population showed the highest dissimilarity of the ITS region sequence with other nine populations of O. corymbosa and two populations of O. diffusa. This consequence makes us further understand the molecular diversification between O. corymbosa and O. diffusa, and help to promote the correct use and safety.

• 동의생리병리학회지
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• 제28권3호
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• pp.310-316
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• 2014

Oldenlandia diffusa, Cremastra appendiculata and Fritillaria thunbergii are widely distributed in the Korea, China and Japan, and has been used in traditional medicine for various diseases, such as pharyngolaryngitis, tonsillitis, goiter and stomach ulcer. However, the anti-cancer activities in human breast cancer have not been clearly elucidated yet. In this study, it was compared the in vitro cytotoxic effects of single and complex treatment of O. diffusa, C. appendiculata and F. thunbergii. We treat human breast cancer MCF-7 cells with O. diffusa, C. appendiculata and F. thunbergii. And we evaluated viability, growth inhibition, morphological changes, apoptotic bodies formation, measurement of the cell cycle and formation of DNA fragmentation of these cells. It was found that single treatment of O. diffusa could inhibit the cell proliferation in human breast cancer MCF-7 cells. However, complex treatment of O. diffusa, C. appendiculata and F. thunbergii is weakly or not affect the cell proliferation of MCF-7 cells. And anti-proliferative effects of O. diffusa in MCF-7 cells was associated with G1 arrest of cell cycle. These findings suggest that O. diffusa may be a potential chemotherapeutic agent for the control of human breast cancer cells and further studies will be needed to identify the molecular mechanisms.

• 대한한방내과학회지
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• 제35권2호
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• pp.133-144
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• 2014

O. diffusa, C. appendiculata and F. thunbergii are reported to possess many pharmacological activities including anti-oxidant, anti-inflammatory, anti-hypertension, anti-diabetic and anti-cancer effects. However, their anti-cancer activities in human breast cancer have not been clearly elucidated yet. Objectives: In the present study, we compared the in vitro cytotoxic effects of single and complex treatment of O. diffusa, C. appendiculata and F. thunbergii in human breast cancer MDA-MB-231 cells. Methods: After we treated human breast cancer MDA-MB-231 cells with O. diffusa, C. appendiculata and F. thunbergii. we evaluated viability, growth inhibition, morphological changes, apoptotic body formation, measurement of the cell cycle and formation of DNA fragmentation of these cells. Results: We found that single treatment of O. diffusa and F. thunbergii could inhibit cell proliferation in human breast cancer MDA-MB-231 cells. However, complex treatment of O. diffusa, C. appendiculata and F. thunbergii had weak or no effect on the cell proliferation of MDA-MB-231 cells. The first, anti-proliferative effects of O. diffusa in MDA-MB-231 cells was associated with G2/M arrest of cell cycle and apoptotic cell death. The second, anti-proliferative effect of F. thunbergii in MDA-MB-231 cells was associated with apoptotic cell death. Conclusions: Taken together, these findings suggest that O. diffusa and F. thunbergii may be a potential chemotherapeutic agent for the control of human breast cancer cells, further studies will be needed to identify the molecular mechanisms.

• 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.

• 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.

• 식물분류학회지
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• 제40권3호
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• pp.145-152
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• 2010

한국산 백운풀속(Oldenlandia L.) 5종에 대하여 외부형태 형질을 재검토하고, 이중 25개의 양적 형질에 대한 수리분류를 통하여 분류군간의 한계를 파악하고자 하였다. 질적 형질에서 잎의 형태, 화서, 화관 내부의 연모, 열매의 상부 팽창, 종자의 형태 등이 분류군 간의 식별형질로 가치가 있었다. 양적 형질의 주성분분석 결과, 주성분 1과 주성분 2를 축으로 하는 산포도에서 산방백운풀(O. corymbosa), 탐나풀(O. hirsuta) 및 낚시돌풀(O. strigulosa)은 뚜렷하게 구분되었으나, 형태적으로 다소 유사한 제주백운풀(O. brachypoda)과 백운풀(O. diffusa)은 다소 겹치게 나타났다. 하지만 두 종은 형태적으로도 꽃받침 열편의 형태, 소화경의 길이, 수술대 및 주두의 길이 등의 차이로 뚜렷히 구분되었다. 또한 유집분석에 의해 두 종이 다르게 유집되는 것으로 나타났다.

• Archives of Pharmacal Research
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• 제28권10호
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• pp.1156-1160
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• 2005

An iridoid glycoside, oldenlandoside III (5) was isolated from the n-butanol fraction of methanol extracts of the aerial parts of Oldenlandia diffusa Roxb. along with six others previously characterized iridoid glycosides; geniposidic acid (1), scandoside (2), feretoside (3), 10-O-ben-zoylscandoside methyl ester (4), asperulosidic acid (6) and deacetylasperulosidic acid (7). Compounds 1, 2, and 7 inhibited LDL-oxidation, and showed $63.3{\pm}2.0,\;62.2{\pm}1.6,\;and\;63.8{\pm}1.5\%$ inhibition, respectively, at a concentration of 20 ${\mu}g/mL$.

• 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.

• 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.

• 식물분류학회지
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• 제40권2호
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• pp.130-133
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• 2010

제주도 서귀포시에서 발견된 Oldenlandia brachypoda DC.를 한반도 미기록종으로 확인하여 보고하고자 한다. 본 종은 속 내의 근연종인 백운풀(Oldenlandia diffusa)에 비해 소화경은 짧고, 돌기가 없으며, 엽액에서 1-3개의 꽃이 피고, 수술대는 0.5-0.8 mm로 길며, 악편은 삼각형, 성숙시 열매의 상부는 팽창하지 않는 점에서 구분된다. 국명은 자생지가 제주도인 점을 고려하여 '제주백운풀'로 명명하였으며, 주요 형질에 대한 도해와 기재, 검색표를 제시하였다.