ABSTRACT
We studied the effect of triterpenoid saponins on the development of free-living stages of Heligmosomoides bakeri, a parasitic nematode of the mouse intestine. We evaluated the effectiveness of oleane-type glucuronides (GlcUAOA) isolated from Calendula officinalis and Beta vulgaris. The rhodamine 123 retention assay was used to detect dysfunctions of the major membrane transporter for xenobiotics, P-glycoprotein (Pgp). Both C. officinals and B. vulgaris GlcUAOA affect the development of the free living stages and function of Pgp in H. bakeri. The GlcUAOA inhibits egg hatching and moulting of larvae and also changes their morphology. These saponin fractions reversed the toxic effect of thiabendazole on the nematode; the function of Pgp was also inhibited.
Subject(s)
ATP Binding Cassette Transporter, Subfamily B, Member 1/pharmacology , Glucuronides/pharmacology , Heligmosomatoidea/drug effects , Saponins/pharmacology , Triterpenes/pharmacology , ATP Binding Cassette Transporter, Subfamily B, Member 1/chemistry , Animals , Beta vulgaris/chemistry , Calendula/chemistry , Glucuronides/chemistry , Heligmosomatoidea/physiology , Larva/drug effects , Saponins/chemistry , Triterpenes/chemistry , Verapamil/pharmacologyABSTRACT
BACKGROUND AND PURPOSE: Growing evidence implicates NF-kappaB as an important contributor to metastasis and increased chemoresistance of melanoma. Here, we report the effects of parthenolide on either untreated, cisplatin- or TNFalpha-treated melanoma cell lines A375, 1205Lu and WM793, exhibiting different levels of constitutive NF-kappaB activity. EXPERIMENTAL APPROACH: Electrophoretic mobility shift assay was used to assess changes in NF-kappaB activity, and real-time PCR to evaluate expression of NF-kappaB-regulated genes. Cell cycle arrest and apoptosis were assessed by flow cytometry. Cell death was also visualized by fluorescence microscopy. Migration was determined by scratch assay and invasiveness by Matrigel assay. KEY RESULTS: Parthenolide suppressed both constitutive and induced NF-kappaB activity in melanoma cells. This was accompanied by down-regulation of cancer-related genes, with NF-kappaB-binding sites in their promoters, including: Bcl-X(L), survivin, cyclin D1, interleukin 8 and matrix metalloproteinase 9. When the various effects of 6 microM parthenolide were compared, apoptosis associated with loss of mitochondrial membrane potential was most efficiently induced in 1205Lu cells, cell cycle arrest in G(0)/G(1) phase was observed in WM793 cells, and high metastatic potential was markedly reduced in A375 cells. These findings not only reflected differences between melanoma cell lines in basal expression of NF-kappaB-regulated genes, but also suggested other parthenolide targets involved in cell cycle progression, migration, invasiveness and survival. CONCLUSIONS: Inhibition of constitutive and therapeutically induced NF-kappaB pathway by parthenolide might be useful in the treatment of melanoma, although the diversity of changes induced in melanoma cells with different genetic backgrounds indicate context-dependent poly-pharmacological properties of this compound.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Melanoma/metabolism , Sesquiterpenes/pharmacology , Apoptosis/drug effects , Cell Cycle/drug effects , Cell Line, Tumor , Cell Movement/drug effects , Cell Proliferation/drug effects , Dose-Response Relationship, Drug , Down-Regulation , Drug Screening Assays, Antitumor/methods , Humans , Melanoma/drug therapy , NF-kappa B/metabolism , Neoplasm Metastasis/drug therapy , Neoplasm Proteins/metabolism , Sesquiterpenes/administration & dosageABSTRACT
The mucosal immune system seems to be an important defence mechanism for fish but the binding of IgM in mucosal organs is poorly described in fish. In this study the gene encoding the polymeric Immunoglobulin Receptor (pIgR) in carp has been isolated and sequenced from a liver cDNA-library and aligned with other species. The pIgR of carp consists of 2 Ig domains, a transmembrane and an intracellular region, together 327 amino acids. In situ hybridisations with sense and anti-sense DIG-labelled pIgR RNA probes were performed on liver, gut and skin of common carp (Cyprinus carpio L.) and in these organs only anti-sense probes were found to hybridise. In liver the majority of hepatocytes was stained around the nucleus. In gut and skin, staining could be detected around the nucleus of the epithelial cells, but in gut also a subpopulation of lymphoid cells was stained in epithelium and lamina propria. The specific in situ hybridisation of the epithelia and hepatocytes coincides with the in situ binding of FITC-labelled carp IgM to the same cells. RT-PCR results indicate the expression of the pIgR gene in all lymphoid organs of carp, but not in muscle. Macrophages/neutrophils enriched by adherence or sorted B cells (MACS) did not show expression of the pIgR gene and are excluded as the pIgR expressing lymphoid cells in the intestine. The relevance of pIgR staining and gene expression in mucosal organs is discussed.
Subject(s)
Carps/genetics , Carps/immunology , Gene Expression Regulation/immunology , Mucous Membrane/immunology , Receptors, Polymeric Immunoglobulin/metabolism , Amino Acid Sequence , Animals , Fluorescein-5-isothiocyanate , Immunoglobulin M/metabolism , In Situ Hybridization, Fluorescence , Molecular Sequence Data , Mucous Membrane/metabolism , Receptors, Polymeric Immunoglobulin/chemistry , Receptors, Polymeric Immunoglobulin/genetics , Reverse Transcriptase Polymerase Chain Reaction , Sequence Alignment , Sequence Analysis, ProteinABSTRACT
A case of a 10-year old child with isolated sixth nerve palsy due to giant intracavernous carotid aneurysm is presented. The aneurysm was confirmed by CT brain scan and carotid angiography. It was closed after local embolization of internal carotid artery with the use of platinum inlays. It turns out that an intracranial aneurysm can be, although very rarely, the cause of isolated sixth nerve palsy in childhood.
