ABSTRACT
p21-activated kinase 1 (Pak1)-a key node protein kinase regulating various cellular process including angiogenesis-has been recognised to be a therapeutic target for multitude of diseases, and hence, various small molecule inhibitors targeting its activity have been tested. However, the direct toxic and anti-angiogenic effects of these pharmacologic agents have not been examined. In this study, we evaluate the translational efficacy of Pak1 inhibitor IPA-3 using zebrafish toxicity model system to stratify its anti-angiogenic potential and off-target effects to streamline the compound for further therapeutic usage. The morphometric analysis has shown explicit delay in hatching, tail bending, pericardial sac oedema and abnormal angiogenesis. We provide novel evidence that Pak1 inhibitor could act as anti-angiogenic agents by impeding the development of sub-intestinal vessel (SIV) and intersegmental vessels (ISVs) by suppressing the expression of vascular endothelial growth factor (VEGF), VEGF receptor 2 (VEGFR2), neurophilin 1 (NRP1) and its downstream genes matrix metalloproteinase (MMP)-2 and MMP-9. Knockdown studies using 2-O-methylated oligoribonucleotides targeting Pak1 also revealed similar phenotypes with inhibition of angiogenesis accompanied with deregulation of major angiogenic factor and cardiac-specific genes. Taken together, our findings indicate that Pak1 signalling facilitates enhanced angiogenesis and also advocated the design and use of small molecule inhibitors of Pak1 as potent anti-angiogenic agents and suggest their utility in combinatorial therapeutic approaches targeting anomalous angiogenesis.
Subject(s)
Angiogenesis Inhibitors/pharmacology , Disulfides/toxicity , Embryo, Nonmammalian/drug effects , Naphthols/toxicity , Toxicity Tests , Zebrafish/embryology , p21-Activated Kinases/antagonists & inhibitors , Animals , Blood Vessels/drug effects , Blood Vessels/embryology , Disulfides/chemistry , Embryo, Nonmammalian/metabolism , Gene Knockdown Techniques , Humans , Models, Animal , Naphthols/chemistry , Neovascularization, Physiologic/drug effects , RNA, Messenger/genetics , RNA, Messenger/metabolism , Vascular Endothelial Growth Factor A/pharmacology , p21-Activated Kinases/metabolismABSTRACT
Silver nanoparticles (Ag NPs) are being increasingly used in many consumer products owing to their excellent antimicrobial properties. The continuous use of Ag NPs in consumer products will lead to environmental release. The present study evaluated the toxic effects and the possible underlying mechanism of Ag NPs on Pseudomonas putida. Ag NP exposure inhibited growth of the cells. Increased lipid peroxidation occurred coincident with suppression of the antioxidant defense system. Ag NP exposure caused reactive oxygen species (ROS) production, glutathione depletion and inactivation of the antioxidant enzyme superoxide dismutase, catalase and glutathione reductase. The addition of superoxide dismutase or pretreatment of P. putida with N-acetyl cysteine that quenches ROS reduced toxicity of the NPs.
