Wj01, A Polyamine Analogue, Attenuate Ischemia Injury In PC12 Cells

Objective: To investigate the protective effect of polyamine analogues (WJ01) on the oxygen-glucose deprivation, glutamate (Glu) and putrescine (Put) induced cytotoxicity in rat pheochromocytoma cells (PC12).Methods: Different models were used to induce the cytotoxicity in PC12 cells, the morphology of PC12 cells was observed by microscope and the cell damage was measured by MTT assay. The reactive oxygen species (ROS) were monitored combined with DCFH-DA by High-Content Screening Reader. DNA-binding dye Acridine Orange (AO) and Ethidium bromide (EB) were used to determine the morphological characteristic of apoptotic cells. Mitochondrial membrane potential (MMP) in PC12 cells were monitored by fluorospectrophotometer combining with Rh123. Fura-2/AM, a cell permeable fluorescent probe, was employed to detect intracellular Ca²⁺ concentration ([Ca²⁺]i).Results: 1. After treatment with 2 mM Na₂S₂O₄ in PC12 cells for 30 min, the cell viability decreased markedly, while WJ01 at the different concentrations (1μM, 5μM, 10μM) could elevate the cell viability obviously. 2. The results suggested that 12.5μM Put decreased the cell viability and the mitochondrial membrane potential, in contrast, Put increased ROS and intracellular [Ca²⁺]i. WJ01 significantly decreased ROS and intracellular [Ca²⁺]i, increased MMP in PC12 cells. 3. After the treatment with 50 mM Glu, the mitochondrial membrane potential significantly decreased while ROS and intracellular [Ca²⁺]i increased, and microscopic observation showed that PC12 cells exhibited morphological alterations such as cell shrinkage and membrane blabbing, the typical characteristics of apoptotic cell death, which was reduced with co-treatment of WJ01. Conclusions: The study shows that WJ01 at the different concentration has a protective effects against Na₂S₂O₄, Put and Glu-induced cytotoxicity and apoptosis in PC12 cells, which may represent the cellular mechanisms including scavenging ROS, increasing MMP, inhibiting the cell apoptosis and overload of Ca²⁺ in PC12 cells. WJ01 may represent a potential treatment strategy for attenuating ischemia injury in PC12 cells.