• Journal of Physiology & Pathology in Korean Medicine
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• v.28 no.3
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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.

• The Journal of Internal Korean Medicine
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• v.35 no.2
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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.

• Korean Journal of Plant Taxonomy
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• v.41 no.1
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• pp.58-65
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• 2011

Commelina benghalensis L. and Commelina diffusa Burm. f. belonging to Commelinaceae, two species previously unrecorded as Korean flora, were collected in the lowlands of Jeju Island in Korea. Commelina benghalensis of the two species is different from others of Commelinaceae in Korea by having funnelform spathes fused at the proximal margin of involucral bracts and cleistogamous flowers. In addition, Commelina diffusa differ from others by having a three-valve capsule and lanceolate and a base cordate or rounded spathelike involucral bracts. The somatic chromosome number of Commelina benghalensis was 2n = 2x = 22 and the sizes of chromosomes were very small, ranging from 1.25 to 2.70 ${\mu}m$. However, the Commelina diffusa chromosome number exceeded 2n = ca. 100, and a precise count could not be obtained. These species are known to be distributed in tropical and subtropical regions of Asia and Africa below the latitude of Jeju Island of Korea.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.6 no.1
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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.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.6 no.1
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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.

• Korean Journal of Plant Taxonomy
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• v.51 no.1
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• pp.10-17
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• 2021

Invasive alien species management is pivotal for biodiversity conservation. Commelina caroliniana Walter, from the family Commelinaceae, is an alien plant native to the Himalayas and India, but it has been widely introduced around the world, including in the United States, Brazil, Philippines, and Japan. In Korea, the first population was found growing adjacent to agricultural land and farm roads on Jejudo Island, and field observations confirmed the presence of at least nine populations there. It is similar morphologically to C. diffusa Burm. f. but can be distinguished by involucral bracts that are ciliate at the base, hairs on the peduncle and obsolete upper cincinnus, brown spots on its 4-lobed antherode, and seed surfaces that are smooth to slightly alveolate. It was determined to have an invasiveness low score of 8 according to the Korean 'Invasive Alien Plant Risk Assessment', suggesting that it may spread to natural habitats. Although the current distribution of C. caroliniana is restricted to Jeju-si, it has spread dramatically in many other areas of the world. At present, it has had a limited impact on the local environment, but local and regulatory authorities should pay close attention to this plant and take measures to prevent its expansion in the future.

• Korean Journal of Soil Science and Fertilizer
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• v.45 no.4
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• pp.602-609
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• 2012

Burkholderia anthina R-4183, Burkholderia diffusa R-15930 and Burkholderia stabilis LMG 14294 isolated from green house soils (Gongju-Gun area, South Korea) were characterized and their phosphate solubilizing ability was assessed. Under in vitro culture conditions, all three species were proved to be effective in solubilizing phosphates in varying degrees. Strain Burkholderia anthina exhibited the highest phosphate solubilization in NBRIP medium ($665{\mu}g\;ml^{-1}$) followed by Burkholderia diffusa ($630{\mu}g\;ml^{-1}$) and Burkholderia stabilis ($578{\mu}g\;ml^{-1}$). However, solubilization of $FePO_4$ and $AlPO_4$ was found to be poor in all the strains. Acidification by means of gluconic and oxalic acids accumulation in the culture medium could be the possible mechanism responsible for phosphate solubilization. Glucose at the rate of 3% was found be the best carbon source for Burkholderia anthina while other two Burkholderia species showed maximum phosphate solubilization at 2% of glucose. In the case of nitrogen sources, ammonium and nitrate were equally effective in solubilizing phosphates by Burkholderia species. Despite a slight decrease in phosphate solubilization observed at increasing temperature, all three Burkholderia species could withstand a temperature of $30-35^{\circ}C$, pH at the range of 7-9 and the presence of NaCl (up to 2.5%) without much compromising the phosphate solubilization. As shown with potted mung bean seedlings, all the three isolates could enhance soil fertility and plant growth indicating their great potential to be used as bio-inoculants.

• International Journal of Industrial Entomology and Biomaterials
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• v.18 no.1
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• pp.49-52
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• 2009

An in vitro and in vivo studies were conducted to evaluate the antibacterial efficacy of certain botanicals viz., rhizomes of turmeric (Curcuma longa) and leaves of amla (Phyllanthus emblica), asparagus (Asparagus racemosus), bael (Aegle marmelos), boerhavia (Boerhavia diffusa), garlic (Allium sativum) and basil (Oscimum basicilum) against bacterial pathogens viz., Staphylococcus sp., Bacillus sp. and Klebsiella cloacae, of silkworm, Bombyx mori. Asparagus and basil, amla and boerhavia, basil and bael at concentration of 20, 000 ppm showed higher antibacterial activity against Staphylococcus sp., Bacillus sp., K. cloacae respectively, both in vitro and in vivo studies.

• Journal of Life Science
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• v.23 no.8
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• pp.1025-1031
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• 2013

From a 95% ethanolic extract of H. diffusa, four marker compounds (HD1~HD4) were isolated, which were relatively unique and exist in comparably high contents. The structures of marker compounds were identified as digitolutein (1), 2-hydroxy-3-methylanthraquinone (2), (E/Z)-6-O-p-coumaroyl scandoside methyl ester (4:1 mixture) (3), and (E/Z)-6-O-p-methoxycinnamoyl scandoside methyl ester (4:1 mixture) (4), respectively, on the basis of $^{13}C$ and $^1H$-NMR analyses. The calibration curves of marker compounds showed high linearity, as their correlation coefficient ($R^2$) were in the range of 0.9991~0.9999. In addition, the limit of detection (LOD) and the limit of quantification (LOQ) were $0.03{\sim}0.07{\mu}g/ml$ and $0.099{\sim}0.231{\mu}g/ml$, respectively. The intra-day/inter-day precision and accuracy were 0.23~2.00%/0.25~1.16% and 94.60~108.44%/94.73-110.23%, respectively. The optimal HPLC conditions for the simultaneous quantification of HD1~HD4 were as follows: stationary phase; Merck Chromolith RP-18e ($100{\times}4.6mm$, $5{\mu}m$), column temp.; room temperature, UV detection at 280 nm, flow rate; 2.0 ml/min, injection volume; $10{\mu}l$, mobile phase; start with the mixture of 80% solvent A ($H_2O$ containing 0.5% acetic acid) and 20% solvent B (methanol containing 0.5% acetic acid) and gradually decrease solvent A to 40% in 9 min., then retain this condition to 18 min. Under the HPLC condition, the four marker compounds 1~4 were successfully separated without any interference of other constituents. The results obtained in this study are expected to be helpful for the development of nutraceutics and natural medicines and for the quality control of this plant.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.2
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• pp.408-412
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• 2004

For the evaluation of immunomodulatory effect of Korean Oldenlandiae Herb (OH) and Radix (OR), our experiment was performed with methanol extracts of Korean Oldenlandiae Herba and Radix. After administration of methanol extracts of Korean OH and OR for 7 days, Balb/C mice were immunized with sheep red blood cells. Four days later, splenic leukocytes were isolated and immunological experiments were performed. Rosette forming cells and plaque forming cells were significantly increased in Korean OH and OR treated mice compared with PBS treated control. Korean OH and OR also enhanced T and B lymphocytes, macrophage and natural killer cells by flowcytometric analysis. LPS-induced TNF-α and IL-6 levels were increased by OH and OR compared with untreated control. These results suggest that Korean OH and OR have immunomodulatory activity through regulation of cell-mediated immune and humoral immune response.