Anticancer activity of gomisin J from Schisandra chinensis fruit.

In attempting to identify effective anticancer drugs from natural products that are harmless to humans, we found that the gomisin J from Schisandra chinensis fruit has anticancer activity. Schisandra chinensis fruits are used in traditional herbal medicine and gomisin J is one of their chemical constituents. In the present study, we examined the anticancer activity of gomisin J in MCF7 and MDA-MB-231 breast cancer cell lines and in MCF10A normal cell line, in a time- and concentration-dependent manner. Our data revealed that gomisin J exerted a much stronger cytotoxic effect on MCF7 and MDA-MB-231 cancer cells than on MCF10A normal cells. Gomisin J suppressed the proliferation and decreased the viability of MCF7 and MDA-MB-231 cells at relatively low (<10 µg/ml) and high (>30 µg/ml) concentrations, respectively. Our data also revealed that gomisin J induced necroptosis, a programmed form of necrosis, as well as apoptosis. Notably, gomisin J predominantly induced necroptosis in MCF7 cells that are known to have high resistance to many pro-apoptotic anticancer drugs, while MDA-MB-231 exhibited a much lower level of necroptosis but instead a higher level of apoptosis. This data indicated the possibility that it may be used as a more effective anticancer drug, especially in apoptosis-resistant malignant cancer cells. In an extended study, gomisin J exhibited a strong cytotoxic effect on all tested various types of 13 cancer cell lines, indicating its potential to be used against a wide range of different types of cancer cells.

Acetyl-cholinesterase Inhibitory Activity of Methoxyflavones Isolated from Kaempferia parviflora.

MeOH. extracts of Kaenpferia parvflora Wall.- ex. Baker family Zingiberaceae; were consecutively partitioned- with CHCl3, EtOAc, nd n-BuOH. The CHC1, fractions were diluted in distilled water with n-hexane-CH2Cl2 and three methoxyflavones were isolated from. the CH2Cl2 extract. Based on-spectral analysis and comparison of the spectral data with literature values, the compounds. were identified as; 3,5,7,3',4'-pentamethoxyflavone (KPl), 5,7- dimethoxyflavone (KP2), and 5,7,4'-trimethoxyflavone (KP3). In relation to their possible effectiveness against Alzheimher's disease;these-compounds were tested for their ability to inhibit acetylcholinesterase activity and neurite outgrowth in the PC12 cell line. Of the three compounds, KPl was the only one to inhibit significantly the acetylcholinesterase activity in a dose-dependent manner.

Protective activity of C-geranylflavonoid analogs from Paulownia tomentosa against DNA damage in 137Cs irradiated AHH-1 cells.

Radiotherapy is an important form of treatment for a wide range of cancers, but it can damage DNA and cause adverse effects. We investigated if the diplacone analogs of P. tomentosa were radio-protective in a human lymphoblastoid cell line (AHH-1). Four geranylated flavonoids, diplacone, 3'-O-methyl-5'-hydroxydiplacone, 3'-O-methyl-5'-O-methyldiplacone and 3'-O-methyldiplacol, were tested for their antioxidant and radio-protective effects. Diplacone analogs effectively scavenged free radicals and inhibited radiation-induced DNA strand breaks in vitro. They significantly decreased levels of reactive oxygen species and cellular DNA damage in 2 Gy-irradiated AHH-1 cells. Glutathione levels and superoxide dismutase activity in irradiated AHH-1 cells increased significantly after treatment with these analogs. The enhanced biological anti-oxidant activity and radioprotective activity of diplacone analogs maintained the survival of irradiated AHH-1 cells in a clonogenic assay. These data suggest that diplacone analogs may protect healthy tissue surrounding tumor cells during radiotherapy to ensure better control of radiotherapy and allow higher doses of radiotherapy to be employed.

Effect of osajin and pomiferin on antidiabetic effects from normal and streptozotocin-induced diabetic rats.

The present study evaluated the antidiabetic effect of osajin and pomiferin from the osajin orange in normal and streptozotocin-induced diabetic rats. Pomiferin in the streptozotocin-induced diabetic effects showed significant hypoglycemic activity for 14 days significantly decreased the serum glucose, triglyceride while it increased the serum insulin in diabetic rats but not in normal rats (p < 0.05; at doses of 100 and 300 mg/kg for 14 days). Pomiferin showed potential in anti-diabetic effects compared to osajin. It also has no effects on C-peptide (ECLIA). Further structure-activity relationships of aromatic position 3 on ring B from osajin and pomiferin will be reported in due course.

Oleifolioside B-mediated autophagy promotes apoptosis in A549 human non-small cell lung cancer cells.

The biochemical mechanisms of cell death by oleifolioside B (OB), a cycloartane-type triterpene glycoside isolated from Dendropanax morbifera Leveille, were investigated in A549 human lung carcinoma cells. Our data indicated that exposure to OB led to caspase activation and typical features of apoptosis; however, apoptotic cell death was not prevented by z-VAD-fmk, a pan-caspase inhibitor, demonstrating that OB-induced apoptosis was independent of caspase activation. Subsequently, we found that OB increased autophagy, as indicated by an increase in monodansylcadaverine fluorescent dye-labeled autophagosome formation and in the levels of the autophagic form of microtubule-associated protein 1 light chain 3 and Atg3, an autophagy-specific gene, which is associated with inhibiting phospho-nuclear factor erythroid 2-related factor 2 (Nrf2) expression. However, pretreatment with bafilomycin A1, an autophagy inhibitor, attenuated OB-induced apoptosis and dephosphorylation of Nrf2. The data suggest that OB-induced autophagy functions as a death mechanism in A549 cells and OB has potential as a novel anticancer agent capable of targeting apoptotic and autophagic cell death and the Nrf2 signaling pathway.

Triptolide-Mediated Apoptosis by Suppression of Focal Adhesion Kinase through Extrinsic and Intrinsic Pathways in Human Melanoma Cells.

Triptolide (TPL) has been shown to inhibit cell proliferation and induce apoptosis in various human cancer cells; however, the precise mechanism of apoptosis induced by TPL in human melanoma cells has not yet been elucidated. In this study, we investigated the precise mechanism underlying cytocidal effects of TPL on human melanoma cells. Treatment of human melanoma cells with TPL significantly inhibited cell growth and induced apoptosis, as evidenced by flow cytometry and annexin V-fluorescein isothiocyanate analyses. TPL increased the levels of Fas and Fas-associated death domain (FADD) and induced cleavage of Bid by activation of caspase-8 and cytochrome c release from mitochondria to the cytosol, which resulted in activation of caspase-9 and caspase-3. Moreover, TPL-induced apoptosis in SK-MEL-2 cells was mediated through dephosphorylation of focal adhesion kinase (FAK) and its cleavage by caspase-8-mediated caspase-3 activation via upregulation of Fas expression. We also found that TPL mediated the dissociation of receptor-interacting protein (RIP) from FAK and enhanced the formation of RIP/Fas complex formation initiating cell death. In conclusion, our data firstly demonstrated that TPL induces apoptosis by both extrinsic and intrinsic apoptosis pathways in human melanoma cells and identified that RIP shuttles between Fas and FAK to mediate apoptosis.

Triptolide induces apoptosis through extrinsic and intrinsic pathways in human osteosarcoma U2OS cells.

Triptolide, a diterpene derived from Tripterygium wilfordii Hook f., a Chinese medicinal herb, has been reported to inhibit cell proliferation and induce apoptosis in various human cancer cells, but its anticancer effects on human osteosarcoma cells have not yet been elucidated. In this study, we investigated whether triptolide induces apoptosis in human osteosarcoma cells and the underlying molecular mechanisms. We firstly demonstrated that triptolide inhibited cell growth and induced apoptosis in U2OS cells. Western blot analysis showed that the levels of procaspase-8, -9, Bcl-2, Bid and mitochondrial cytochrome c were downregulated in triptolide-treated U2OS cells, whereas the levels of Fas, FasL, Bax, cytosolic cytochrome c, cleaved caspase-3 and cleaved PARP were upregulated. These results suggest that triptolide induces apoptosis in U2OS cells by activating both death receptor and mitochondrial apoptotic pathways.

Neuroprotective effects of triterpene glycosides from glycine max against glutamate induced toxicity in primary cultured rat cortical cells.

To examine the neuroprotective effects of Glycine max, we tested its protection against the glutamate-induced toxicity in primary cortical cultured neurons. In order to clarify the neuroprotective mechanism(s) of this observed effect, isolation was performed to seek and identify active fractions and components. From such fractionation, two triterpene glycosides, 3-O-[α-l-rhamnopyranosyl(1-2)-ß-d-glucopyranosyl(1-2)-ß-d-glucuronopyranosyl]olean-12-en-3ß,22ß,24-triol (1) and 3-O-[ß-d-glucopyranosyl(1-2)-ß-d-galactopyranosyl(1-2)-ß-d-glucuronopyranosyl]olean-12-en-3ß,22ß,24-triol (2) were isolated with the methanol extracts with of air-dried Glycine max. Among these compounds, compound 2 exhibited significant neuroprotective activities against glutamate-induced toxicity, exhibiting cell viability of about 50% at concentrations ranging from 0.1 µM to 10 µM. Therefore, the neuroprotective effect of Glycine max might be due to the inhibition of glutamate-induced toxicity by triterpene glycosides.

Compounds with antiproliferative activity on five human cancer cell lines from South Korean Carpesium triste.

In the search for antiproliferative compounds against human cancer cells (A549, SK-OV-3, SK-MEL-2, XF498, HCT15), it was found that the chloroform extracts obtained from the whole plant of Carpesium rosulatum Miq. (Compositae) exhibited significant cytotoxic activity. Two sesquiterpene lactones, CT-1 (2alpha,5-epoxy-5,10-dihydroxy-6-angeloyloxy-9beta-isobutyloxy-germacran-8alpha, 12-olide), and CT-2 (2beta,5-epoxy-5,10-dihydroxy-6alpha,9beta-diangeloyloxy-germacran-8alpha,12-olide) were isolated from the whole plant. CT-2 showed the most potent cytotoxicity with an IC50 value of 7.88 microM against SK-MEL-2.

Oleifolioside A, a New Active Compound, Attenuates LPS-Stimulated iNOS and COX-2 Expression through the Downregulation of NF-κB and MAPK Activities in RAW 264.7 Macrophages.

Oleifolioside A, a new triterpenoid compound isolated from Dendropanax morbifera Leveille (D. morbifera), was shown in this study to have potent inhibitory effects on lipopolysaccharide (LPS-)stimulated nitric oxide (NO) and prostaglandin E(2) (PGE(2)) production in RAW 264.7 macrophages. Consistent with these findings, oleifolioside A was further shown to suppress the expression of LPS-stimulated inducible nitric oxide synthase (iNOS) and cyclooxigenase-2 (COX-2) in a dose-dependent manner at both the protein and mRNA levels and to significantly inhibit the DNA-binding activity and transcriptional activity of NF-κB in response to LPS. These results were found to be associated with the inhibition of the degradation and phosphorylation of IκB-α and subsequent translocation of the NF-κB p65 subunit to the nucleus. Inhibition of NF-κB activation by oleifolioside A was also shown to be mediated through the prevention of p38 MAPK and ERK1/2 phosphorylation. Taken together, our results suggest that oleifolioside A has the potential to be a novel anti-inflammatory agent capable of targeting both the NF-κB and MAPK signaling pathways.

Oleifolioside A mediates caspase-independent human cervical carcinoma HeLa cell apoptosis involving nuclear relocation of mitochondrial apoptogenic factors AIF and EndoG.

Apoptosis, the main type of programmed cell death, plays an essential role in a variety of biological events. Whereas "classical" apoptosis is dependent on caspase activation, caspase-independent death is increasingly recognized as an alternative pathway. To develop new anticancer agents, oleifolioside A was isolated from Dendropanax morbifera Leveille and the biochemical mechanisms of oleifolioside A-induced apoptosis in HeLa cells were investigated. Exposure to oleifolioside A resulted in caspase activation and typical features of apoptosis, although cell death was not prevented by caspase inhibition. Oleifolioside A treatment induced up-regulation of Bad, loss of mitochondrial membrane potential, nuclear relocation of mitochondrial factors, apoptosis-inducing factor (AIF), endonuclease G (EndoG), and apoptosis induction. This is the first report of anticancer activity of oleifolioside A, and nuclear translocation of AIF and EndoG in oleifolioside A-treated HeLa cells might represent an alternative death signaling pathway in the absence of caspase activity.

Inhibitory effect and mechanism on antiproliferation of isoatriplicolide tiglate (PCAC) from Paulownia Coreana.

Paulownia coreana has traditionally been used as the medicine and health food in the treatment of cancer and infectious diseases. In the present study, a new antiproliferation agent, isoatriplicolide tiglate (PCAC) was isolated from the chloroform soluble fraction of the leaves of Paulownia coreana. The antiproliferation activities of PCAC plant extract was examined in breast and cervical cancer cell lines in a time-and dose-dependent manners. Our in vitro experiments showed that PCAC suppresses the cell growth and proliferation of cancer cells at a relatively low concentration (< 10 µg/mL) and induces apoptosis at a high concentration (> 50 µg/mL). Western blot analysis showed that concentration higher than 50 µg/mL induces a time-dependent increase in the percentage of apoptotic cells. In this case, PCAC uses both extrinsic and intrinsic pathways for the apoptosis. PCAC treatment decreased the expression of pro-caspase 8, 9, and 3, the main regulators of apoptotic cell death, in MDA-MB-231 cells, accompanied by the activation of caspase 8, 9, and 3. More importantly, PCAC inhibited the in vitro proliferation of six other human breast and cervical cancer cell lines. In conclusion, our data strongly suggest that PCAC acts as an antiproliferation agents particularly against breast and cervical cancers by inducing cell cycle arrest in the S/G2 phase and caspase dependent apoptosis at relatively low (< 10 µg/mL) and high (> 50 µg/mL) concentrations, respectively.

JNP3, a new compound, suppresses PMA-induced tumor cell invasion via NF-κB down regulation in MCF-7 breast cancer cells.

The expression of matrix metalloproteinase (MMPs)-9 is critical for cell migration and can lead to invasion and metastasis of cancer cells. In the present study, we examined the inhibitory effects of JNP3, a new compound which was isolated from traditional Chinese medicine, on cell invasion and MMP-9 activation in phorbol myristate acetate (PMA)-induced MCF-7 cells. Treatment with JNP3 significantly and selectively inhibited PMA-induced MMP-9 secretion, mRNA expression and protein levels, and these results led to reduction of cell invasion and migration in PMA-induced MCF-7 cells. The results of MMP-9 promoter assay and EMSA showed that JNP3 specifically inhibited PMA-induced MMP-9 gene expression by blocking NF-κB-dependent transcriptional activity. In addition, PMA-induced phosphorylation of ERK1/2 and JNK were suppressed by JNP3 treatment, whereas the phosphorylation of p38 MAPK was not affected by JNP3. These results suggest that JNP3 can be potential anti-cancer agents through specific inhibition of NF-κB-dependent MMP-9 gene expression.

Anticomplement activity of various solvent extracts from Korea local Artemisia spp.

The study evaluated the anticomplement activity from various solvent extracts of eight Artemisia plants (Artemisia capillaris Thunb., Artemisia fukudo Makino., Artemisia japonica Thunb., Artemisia montana (Nakai) Pamp., Artemisia keiskeana Miq., Artemisia rubripes Nakai., Artemisia stolonifera (Maxim.) Kom., and Artemisia sylvatica Max.) from South Korea on the classical pathway (CP). We have evaluated various organic solvent extract from eight Artemisia plants with regard to its anticomplement activity on the CP. A. rubripes and A. montana chloroform extracts showed inhibitory activity against complement system with 50% inhibitory concentrations (IC50) values of 54.3 and 64.2 µg/mL. This is the first report of anticomplement activity from Artemisia plants.

Anticomplement activity of isolated compounds from Artemisia montana.

The study evaluated the anticomplement activity from isolated compounds from Artemisia montana (Nakai) Pamp. from South Korea on the classical pathway. In a previous work, A. montana (Nakai) Pamp. chloroform extracts showed inhibitory activity against complement system. The chromatographic separation of a chloroform chloride extract of A. montana (Nakai) Pamp. led to the isolation of four compounds. Their structures were characterized to be ezoartemin, yamayomoginin, ezomontanin and 11,13-dihydroezomontanin by spectroscopic data. This is the first report of anticomplement activity of isolated compounds from A. montana (Nakai) Pamp.

Anticomplement activity of organic solvent extracts from Korea local Amarantaceae spp.

The study evaluated the anticomplement activity from various solvent extracts of nine Amarantaceae plants (Achyranthes japonica (Miq.) Nakai, Amaranthus mangostanus L., Amaranthus retroflexus L., Amaranthus spinosus L., Celosia argentea var. spicata., Amaranthus lividus L., Celosia cristata L., Amaranthus viridis L., Gomphrena globosa L.) from South Korea on the classical pathway. We have evaluated various organic solvent extract from nine Amarantaceae plants with regard to its anticomplement activity on the classical pathway. Achyranthes japonica chloroform extracts showed inhibitory activity against complement system with 50% inhibitory concentrations (IC(50)) value of 73.1µg/ml. This is the first report of anticomplement activity from Amarantaceae plants.

Inhibitory effects of three oleanolic acid glycosides from Achyranthes japonica on the complement classical pathway.

The present study evaluated the anticomplement effects of isolated compounds from Achyranthes japonica in the classical pathway of the complement system. Using column chromatography, three compounds: achyranthoside C dimethyl ester, achyranthoside C butyl dimethyl ester, and achyranthoside E dimethyl ester were isolated and evaluated for in vitro anticomplement activity. Achyranthoside C dimethyl ester showed the most potent inhibitory activity against the complement system, with 50% inhibitory concentrations (IC(50)) values of 26.2 µg/mL. This is the first report of anticomplement activity of isolated compounds from A. japonica.

Inhibitory effects of organic solvent extracts from Korean local plants on the complement classical pathway.

The study evaluated the anticomplement effects from organic solvent extracts of five Compositae plants (Ligularia fischeri (Ledeb.) Turez, Ligularia taquetii (H.Lev. & Vaniot) Nakai, Ainsliaea acerifolia Sch.Bip, Aster scaber Thunb, Aster koraiensis Nakai, Synurus deltoides Aiton) from South Korea on the classical pathway complement system. We have evaluated organic solvent extracts from five Compositae with regard to its anticomplement activity. Chloroform extracts from L. taquetii showed inhibitory activity against complement system with 50% inhibitory concentrations (IC50) values of 73.2 µg/mL. This is the first report of anticomplement activity from L. taquetii.

Inhibition effects of isolated compounds from Artemisia rubripes Nakai of the classical pathway on the complement system.

The study evaluated the anticomplement activity from isolated compounds from Artemisia rubripes Nakai from South Korea on the classical pathway. In the previous works, Artemisia rubripes chloroform extracts showed inhibitory activity against complement system. The chromatographic separation of a chloroform chloride extract of Artemisia rubripes led to the isolation of three compounds. Their structures were characterized to be scopoletin (1), 11,(13)-triene-6,12-olide (2), and 1ß,6α-dihydroxy-4(15)-eudesmene (3) by spectroscopic data. This is the first report of anticomplement activity of isolated compounds from Artemisia rubripes.

Isolated compounds from Sorghum bicolor L. inhibit the classical pathway of the complement.

The present study evaluated the anticomplement effects from isolated compounds of Sorghum bicolor in classical pathway complement system. Using column chromatograph, three compounds; Sorgoleone-362 (1), Sorgoleone-360 (2) and Sorgoleone-386 (3) were isolated and evaluated for in vitro anticomplement activity. Sorgoleone-386 showed inhibitory activity against complement system with 50% inhibitory concentrations (IC(50)) values of 148.3µg/ml. This is the first report of anticomplement activity of isolated compounds from Sorghum bicolor.