Cystatin C as a potential therapeutic mediator against Parkinson's disease via VEGF-induced angiogenesis and enhanced neuronal autophagy in neurovascular units
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Abstract
Cystatin C (CYS C, Cst3) is an endogenous cysteine protease inhibitor that plays neuroprotective roles in neurodegenerative diseases. We aimed to explore the association of CYS C with Parkinson's disease (PD) models and investigate its involvement in the role of neurovascular units (NVUs) in PD neuro-pathogenesis. We used A53T alpha-synuclein (SNCA) transgenic mice and 6-hydroxydopamine-lesioned DAergic PC12 cells as experimental PD models to investigate the mechanisms behind this association. The injections of CYS C were administered to the right substantia nigra (SN) of A53T SNCA transgenic mice to measure the effects of CYS C in transgenic A53T SNCA mice. To explore the angiogenesis in vivo and in vitro, we used the chick embryo chorioallantoic membrane (CAM) assay and tube formation (TF) assay. We found that CYS C has a neuroprotective effect in this in vivo PD model. We observed increased VEGF, NURR1 and autophagy markers LC3B and decreased SNCA and apoptosis marker cleaved CASP3 in different brain regions of CYS C-treated A53T SNCA transgenic mice. In vitro, we observed that CYS C-induced VEGF, a secreted protein, attenuated 6-OHDA-lesioned DAergic PC12 cell degeneration by regulating p-PKC-alpha/p-ERK1/2-Nurr1 signaling and inducing autophagy. VEGF-mediated angiogenesis was markedly enhanced in the conditioned media of 6-OHDA-lesioned PC12 cells with CYS C-overexpression, whereas blockage of autophagy in CYS C-overexpressing PC12 cells significantly downregulated VEGF expression and the associated angiogenesis. Our data indicate that CYS C displays dual neuronal-vascular functions, promoting PC12 cell survival and angiogenesis via regulating the level of secreted VEGF in NVUs. Our study provides evidence that may aid in the development of an alternative approach for the treatment of PD through modulation of CYS C-mediated neuronal-vascular pathways.
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Autophagy / drug effects
Caspase 3 / genetics
Caspase 3 / metabolism
Chick Embryo
Chorioallantoic Membrane / blood supply
Chorioallantoic Membrane / drug effects
Chorioallantoic Membrane / metabolism
Cystatin C / pharmacology
Disease Models, Animal
Gene Expression Regulation
Humans
Injections, Intraventricular
Mice
Mice, Transgenic
Microtubule-Associated Proteins / genetics
Microtubule-Associated Proteins / metabolism
Neovascularization, Physiologic / drug effects
Neurons / drug effects
Neurons / metabolism
Neurons / pathology
Neuroprotective Agents / pharmacology
Nuclear Receptor Subfamily 4, Group A, Member 2 / genetics
Nuclear Receptor Subfamily 4, Group A, Member 2 / metabolism
Oxidopamine / antagonists & inhibitors
Oxidopamine / pharmacology
PC12 Cells
Parkinson Disease / genetics
Parkinson Disease / metabolism
Parkinson Disease / pathology
Rats
Signal Transduction
Substantia Nigra / drug effects
Substantia Nigra / metabolism
Substantia Nigra / pathology
Vascular Endothelial Growth Factor A / genetics
Vascular Endothelial Growth Factor A / metabolism
alpha-Synuclein / genetics
alpha-Synuclein / metabolism