ABSTRACT
Autosomal dominant polycystic kidney disease (ADPKD) is characterized by progressive enlargement of kidney cysts, leading to chronic kidney disease. Since the available treatment for ADPKD is limited, there is emerging interest for natural compounds as potential therapeutic candidates. The aim of our study was to investigate whether an olive leaf extract may be able to counteract the cyst growth in an in vitro model of ADPKD. We treated WT9-12 cells with an olive leaf extract (OLE). In monolayer culture we evaluated cell viability by the MTT assay, protein expression by western-blot analysis and apoptosis by DNA laddering and TUNEL assays. For functional studies we used transient transfection and ChIP assays. Intracellular calcium measurement was performed with a spectrofluorimeter using a fluorescent probe. 3D-cell-culture was used for cyst growth studies. OLE reduced the WT9-12 cell growth rate and affected intracellular signaling due to high c-AMP levels, as OLE reduced PKA levels, enhanced p-AKT, restored B-Raf-inactivation and down-regulated p-ERK. We elucidated the molecular mechanism by which OLE, via Sp1, transactivates the p21WAF1/Cip1 promoter, whose levels are down-regulated by mutated PKD1. We demonstrated that p-AKT up-regulation also played a crucial role in the OLE-induced anti-apoptotic effect and that OLE ameliorated intracellular calcium levels, the primary cause of ADPKD. Finally, using a 3D-cell-culture model we observed that OLE reduced the cyst size. Therefore, multifaceted OLE may be considered a new therapeutic approach for ADPKD treatment.
Subject(s)
Cell Proliferation/drug effects , Cysts/prevention & control , Olea/chemistry , Plant Extracts/pharmacology , Plant Leaves/chemistry , Polycystic Kidney, Autosomal Dominant/drug therapy , Apoptosis/drug effects , Cell Line, Tumor , Chromatin Immunoprecipitation , Humans , In Situ Nick-End Labeling , Inhibitory Concentration 50 , Iridoid Glucosides , Iridoids/pharmacology , Mitogen-Activated Protein Kinases/genetics , Mitogen-Activated Protein Kinases/metabolism , Promoter Regions, GeneticABSTRACT
Melanomas are neoplasms not frequent in cattle. They constitute less than 2 per cent of all the bovine tumors. This report describes a 11-month-old female Holstein heifer bearing a subcutaneous mass of tissue approximately 18x8cm surrounding the right eye, extending to the top of the neck. This mass appeared when the animal was 4-mounth-old. A biopsy sample taken to histopathology revealed neoplastic tissue composed of melanin-loaded cells and not disorganized connective tissue fibers. Partial pigment removal, utilizing potassium permanganate technique, revealed large nucleus in the melanocitic cells. In some of these cells, a nucleolus was evident. Considering that previous reports on fibrous dermic melanocytoma are rare in the literature, these findings are documented
