Molecular identification of medicinal herbs, Oldenlandia diffusa and Oldenlandia corymbosa based on nrDNA ITS region sequence

Sun, Yan-Lin,Wang, Dong,Yeom, Myung-Hun,Kim, Duck-Hee,Kim, Han-Gon,Hong, Soon-Kwan The Korean Society of Plant Biotechnology 2011 식물생명공학회지 Vol.38 No.4

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.

Molecular identification of medicinal herbs, Oldenlandia diffusa and Oldenlandia corymbosa based on nrDNA ITS region sequence

Yan-Lin Sun,Dong Wang,Myung-Hun Yeom,Duck-Hee Kim,Han-Gon Kim,홍순관 한국식물생명공학회 2011 식물생명공학회지 Vol.38 No.4

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.

Ethanol Extract of Oldenlandia diffusa - an Effective Chemotherapeutic for the Treatment of Colorectal Cancer in Humans -Anti-Cancer Effects of Oldenlandia diffusa-

Lee, Soojin,Shim, Ji Hwan,Gim, Huijin,Park, Hyun Soo,Kim, Byung Joo KOREAN PHARMACOPUNCTURE INSTITUTE 2016 Journal of pharmacopuncture Vol.19 No.1

Objectives: Oldenlandia diffusa is traditionally used to relieve the symptoms of and to treat various diseases, but its anti-cancer activity has not been well studied. In the present study, the authors investigated the anti-cancer effects of an ethanol extract of Oldenlandia diffusa (EOD) on HT-29 human adenocarcinoma cells. Methods: Cells were treated with different concentrations of an EOD, and cell death was assessed by using a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. Analyses of the sub G1 peak, the caspase-3 and -9 activities, and the mitochondrial membrane depolarizations were conducted to confirm cell death by apoptosis. Also, intracellular reactive oxygen species (ROS) generation was determined using carboxy-H2DCFDA (5-(and-6)-carboxy-20,70-dichlorodihydrofluorescein diacetate). Results: EOD inhibited the proliferation of HT-29 cells for 24 hours by $78.6%{\pm}8.1%$ at $50{\mu}g/mL$, $74.4%{\pm}4.6%$ at $100{\mu}g/mL$, $65.9%{\pm}5.2%$ at $200{\mu}g/mL$, $51.4%{\pm}6.2%$ at $300{\mu}g/mL$, and by $41.7%{\pm}8.9%$ at $400{\mu}g/mL$, and treatment for 72 hours reduced the proliferation at the corresponding concentrations by $43.3%{\pm}8.8%$, $24.3{\pm}5.1mV$, $13.5{\pm}3.2mV$, $6.5{\pm}2.3mV$, and by $2.6{\pm}2.3mV$. EOD increased the number of cells in the sub-G1 peak in a dose-dependent manner. The mitochondrial membrane depolarization was elevated by EOD. Also, caspase activities were dose-dependently elevated in the presence of EOD, and these activities were repressed by a pan-caspase inhibitor (zVAD-fmk). The ROS generation was significantly increased by EOD and N-acetyl-L-cysteine (NAC; a ROS scavenger) remarkably abolished EOD-induced cell death. In addition, a combination of sub-optimal doses of EOD and chemotherapeutic agents noticeably suppressed the growth of HT-29 cancer cells. Conclusion: These results indicate that EOD might be an effective chemotherapeutic for the treatment of human colorectal cancer.

Ethanol Extract of Oldenlandia diffusa – an Effective Chemotherapeutic for the Treatment of Colorectal Cancer in Humans -Anti-Cancer Effects of Oldenlandia diffusa-

이수진,심지환,김희진,박현수,김병주 대한약침학회 2016 Journal of pharmacopuncture Vol.19 No.1

Objectives: Oldenlandia diffusa is traditionally used to relieve the symptoms of and to treat various diseases, but its anti-cancer activity has not been well studied. In the present study, the authors investigated the anti-cancer effects of an ethanol extract of Oldenlandia diffusa (EOD) on HT-29 human adenocarcinoma cells. Methods: Cells were treated with different concentrations of an EOD, and cell death was assessed by using a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. Analyses of the sub G1 peak, the caspase-3 and -9 activities, and the mitochondrial membrane depolarizations were conducted to confirm cell death by apoptosis. Also, intracellular reactive oxygen species (ROS) generation was determined using carboxy-H2DCFDA (5-(and-6)-carboxy-20,70-dichlorodihydrofluorescein diacetate). Results: EOD inhibited the proliferation of HT-29 cells for 24 hours by 78.6% ± 8.1% at 50 μg/mL, 74.4% ± 4.6% at 100 μg/mL, 65.9% ± 5.2% at 200 μg/mL, 51.4% ± 6.2% at 300 μg/mL, and by 41.7% ± 8.9% at 400 μg/mL, and treatment for 72 hours reduced the proliferation at the corresponding concentrations by 43.3% ± 8.8%, 24.3 ± 5.1 mV, 13.5 ± 3.2 mV, 6.5 ± 2.3 mV, and by 2.6 ± 2.3 mV. EOD increased the number of cells in the sub-G1 peak in a dose-dependent manner. The mitochondrial membrane depolarization was elevated by EOD. Also, caspase activities were dose-dependently elevated in the presence of EOD, and these activities were repressed by a pan-caspase inhibitor (zVAD-fmk). The ROS generation was significantly increased by EOD and N-acetyl-L-cysteine (NAC; a ROS scavenger) remarkably abolished EOD-induced cell death. In addition, a combination of sub-optimal doses of EOD and chemotherapeutic agents noticeably suppressed the growth of HT-29 cancer cells. Conclusion: These results indicate that EOD might be an effective chemotherapeutic for the treatment of human colorectal cancer.

Ethanol Extract of <i>Oldenlandia diffusa</i> Herba Attenuates Scopolamine-Induced Cognitive Impairments in Mice via Activation of BDNF, P-CREB and Inhibition of Acetylcholinesterase

Lee, Jung Eun,Song, Hyo-Sook,Park, Moon Nyeo,Kim, Sung-Hoon,Shim, Bum-Sang,Kim, Bonglee MDPI 2018 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.19 No.2

<P>Though <I>Oldenlandia diffusa</I> Herba (ODH) has been known to exhibit anti-cancer and anti-inflammatory effects, its anti-amnestic effect has never been reported so far. The aim of this present study was to elucidate the anti-amnestic effect of ODH. ODH pretreatment significantly reduced escape latency of scopolamine treated Institute of Cancer Research (ICR) mice compared to untreated control groups in a Morris water maze test. Similarly, the passive avoidance test showed that ODH treatment recovered the scopolamine induced amnesia in the ICR mouse model. Concentration of Ach in brains of ODH treated mice was increased compared to that of scopolamine treated mice. In addition, activity of acetylcholinesterase (AChE) was notably decreased by ODH. The protein expression of brain-derived neurotrophic factor (BDNF) and phospho-cAMP response element-binding protein (p-CREB) (Ser133) was increased in ODH pretreated group compared to control group. Consistently, immunohistochemistry (IHC) revealed the elevated expression of brain-derived neurotrophic factor (BDNF) and p-CREB in brains of ODH treated mice compared to the control group. Overall, these findings suggest that ODH has anti-amnestic potential via activation of BDNF and p-CREB and inhibition of AChE in mice with scopolamine induced amnesia.</P>

백화사설초 추출물의 인체 대장암 세포주에서 항암효능에 관한 연구

이수진,김희진,심지환,박현수,김병주,Lee, Soojin,Gim, Huijin,Shim, Ji Hwan,Park, Hyun Soo,Kim, Byung Joo 대한한의학방제학회 2015 大韓韓醫學方劑學會誌 Vol.23 No.1

Objectives : The purpose of this study was to investigate the anti-cancer effects of Oldenlandia diffusa extract on WiDr human colorectal adenocarcinoma cells. Methods : We examined cell death by (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) MTT assay and the caspase 3 and 9 activity assay with Oldenlandia diffusa extract. To examine the inhibitory effects of Oldenlandia diffusa extract, we performed a cell cycle (sub-G1) analysis and mitochondrial membrane potential for the WiDr cells after 24 hours with Oldenlandia diffusa extract. Results : 1. Oldenlandia diffusa extract induced cell death in WiDr cells. 2. The sub-G1 peak was increased by Oldenlandia diffusa extract in WiDr cells. 3. Oldenlandia diffusa extract leads to increase the mitochondrial membrane depolarization in WiDr cells. 4. Oldenlandia diffusa extract increases caspase 3 and 9 activities in WiDr cells. 5. Oldenlandia diffusa extract combined with several anti-cancer drugs (paclitaxel, 5-fluorouracil, cisplatin, ectoposide, doxorubicin and docetaxel) markedly inhibited the growth of WiDr cells compared to Oldenlandia diffusa extract and anti-cancer drugs alone. Conclusions : Oldenlandia diffusa extract has an apoptotic role in human colorectal cancer cells and a potential role in developing therapeutic agents against colorectal cancer.

Iridoid Glycosides Isolated from Oldenlandia diffusa Inhibit LDL-Oxidation

Dong-Hyun Kim,Hyo-Jung Lee,Young-Jun Oh,Min-Jung Kim,Sung-Hoon Kim,Tae-Sook Jeong,Nam-In Baek 대한약학회 2005 Archives of Pharmacal Research Vol.28 No.10

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-benzoylscandoside methyl ester (4), asperulosidic acid (6) and deacetylasperulosidic acid (7). Compounds 1, 2, and 7 inhibited LDL-oxidation, and showed 63.3±2.0, 62.2±1.6, and 63.8±1.5% inhibition, respectively, at a concentration of 20 µg/mL.

백화사설초 메탄올 추출물이 acetaminophen으로 유도된 생쥐의 급성 간손상에 대한 효능 연구

김종대,문진영,Kim, Jong-Dae,Moon, Jin-Young 대한한의학방제학회 2001 大韓韓醫學方劑學會誌 Vol.9 No.1

Objectives : Oldenlandiae Diffusae herba has been used as a natural drug for tumor, inflammation and liver disease in traditional medicine. This study was performed in order to investigate the antioxidative effects of Oldenlandiae Diffusae herba methanol extract(ODHM) on acetaminophen induced acute liver injury in mice. Methods : In order to investigate the protective effect of ODHM on acute hepatic injury in vivo, ICR mice were pretreated with ODHM, and then treated with acetaminophen(500mg/kg). And the levels of LPO and glutathione(GSH), antioxidative enzyme activities were measured. The levels of LPO were measured by TBA method. And catalase activity was measured as the decrease in hydrogen peroxide absorbance at 240nm on spectrophotometer using 30mM hydrogen peroxide. Superoxide dismutase(SOD) was assayed by recording the inhibition of nitro blue tetrazolium reduction with xanthine and xanthine oxidase. Glutathione peroxidase(GPX) activity was determined by the modified coupled assay developed by Paglia and Lawrence. The reaction was started by addition of 2.2mM hydrogen peroxide as substrate. The change in absorbance at 340nm was measured for 1min on spectrophotometer. Glutathione-S-transferase(GST) activity was assayed with CDNB as substrate and enzyme activity of GST towards the glutathione conjugation of CDNB. And Total SH and GSH levels were measured. Results : In vivo study, LPO levels of acetaminophen treatment group were significantly higher than other groups. This increased level was significantly reduced by ODHM pretreatment. The acetaminophen treatment resulted in a decrease of catalase, GPX, SOD and GST activities. By contrast, ODHM pretreatment markedly increased compare to those of untreated groups. Total SH and GSH levels were reduced by of acetaminophen treatment, and ODHM pretreatment significantly increased GSH levels.

귀전우, 백화사설초, 와송 추출물을 처치한 난소암과 자궁경부암 세포에서의 단백질 발현 변화

김경순,예성철,유병철,조종관,이연월,유화승,Kim, Kyung-Soon,Yea, Sung-Chul,Yoo, Byong-Chul,Cho, Chong-Kwan,Lee, Yeon-Weol,Yoo, Hwa-Seung 대한한방내과학회 2011 大韓韓方內科學會誌 Vol.32 No.1

Background : Despite recent advances in cancer management, prognosis of ovarian cancer is poor. Anticancer effects of herbal medicine, such as Euonymus alatus Sieb, Oldenlandia diffusa (Willd.) Roxburgh, and Orostachys japonicus A. Berger, have been reported in treatment of ovarian and cervical cancers, but the systematic approaches to explain their molecular mechanism(s) have not yet been established. Objectives : To establish a basis of understanding for anti-cancer mechanisms of herbal medicine, we profiled protein expression in human ovarian and cervical cancer cells treated with the extracts from Euonymus alatus Sieb, Oldenlandia diffusa (Willd.) Roxburgh and Orostachys japonicus A. Berger. Methods : Human ovarian cancer cell line NIH:OVCAR-3, and human cervical cancer cell line HeLa were employed in the present study. Whole protein was obtained from the cells harvested at 48 hours after the treatment with herbal water-extract, and analyzed by 2DE-based proteomic approach. Results : Various changes of protein expression induced by the herbal treatment were monitored : down-regulation of molecular chaperone (calreticulin variant), glycolytic enzymes (D-3-phosphoglycerate dehydrogenase, glyceraldehyde 3-phosphate dehydrogenase and alpha-enolase), RNA processing molecules (hnRNP A2/B1), and antioxidant protein (peroxiredoxin 1). Conclusions : Repression of glycolysis has been accepted as the mechanism to increase anticancer reagent's effect. Thus, down-regulation of glycolytic enzymes by the herbal extracts suggested a possible synergistic effect of herbs in the presence of platinum-based therapeutics. In further study, as well as the synergistic effect of the herbs, it has to be further validated whether artificial regulation of hnRNP A2/B1 in ovarian cancer cells affects various cancer survival factors, since RNA processing can be interrupted by deranged expression of hnRNP subtypes, and it results in an inhibition of cancer cell growth.

EFFECT OF THE METHANOL EXTRACT OF KOREAN TRADITIONAL MEDICINE HERB OLDENLANDIAE DIFFUSAE HERBA ON PROLIFERATION AND DIFFERTATION OF CANCER CELL

Jeon,Byung-Hun,Moon,Gu,Woo,Won-Hong 대한동의병리학회 1998 동의생리병리학회지 Vol.12 No.2

백화사설초는 여러 가지 암세포의 성장을 억제할 수 있는 항암효과가 뛰어난 약재로, 근래에 많이 암환자에 활용되어지고 있으며, 실제 실험적으로도 그 효과가 입증되어지고 있다. 본 연구에서는 백화사설초가 사람 암세포의 증식과 분화에 어떠한 효과를 미치는지를 알아보기 위하여 monocytic leukemic cell line인 U937을 백화사설초 추출물만을 처리하고, 또한 백화사설초(10 ㎍/ml)의 농도에서 활성화된 단핵세포(ODH-MNC-CM)를 사용하였다. 그 결과 백화사설초 단독으로 사용할 경우 암세포의 분화를 유도하는 작용은 가지고 있지 않았지만 백화사설초가 Leukemic U937 세포에 대한 증식억제와 분화를 유도하는 IFN-γ, TNF-α와 같은 cytokine의 생산을 야기할 수 있다는 것을 알았으며, 백화사설초 추출물의 항종양효과와 암세포분화효과는 이러한 cytokines의 생성과 관련된 것으로 생각된다. Oldenlandiae diffusae Herba is herb medicine with antitumor activity capable of suppressing the growth of some cancer cell lines. In the present study, we have isolated a methanol extraction of Oldenlandiae diffusae Herba(ODH) and investigated its effect on the proliferation and differentiation of human cancer cell line using an in vitro culture system. Our results showed that the conditioned medium from ODH(10 ㎍/ml)-stimulated blood mononuclear cells (ODH-MNC-CM) treatment induced about 50% of the cells differentiating into mature monocytes/macrophages of CD 11b, CD 14, and CD68. Yet, the differentiated U937 cells also had functions of phagocytosis and superoxide production. However, ODH alone or normal MNC-CM had no such effects. The levels of interferon (IFN)-γ, tumor necrosis factor (TNF)-α, and interleukin (IL)-1 were very low in normal MNC-CM, and they were greatly increased in MNC-CM prepared with ODH stimulation. Antibody neutralization studies further revealed that the tumoricidal and differentiating effects of ODH-MNC-CM were mainly derived from the elevated cytokines, especially IFN-γ and TNF-α. These two cytokines acted synergistically on inhibiting cell growth and inducing differentiation of the target U937 cells.