Construction of PLGA/JNK3-shRNA nanoparticles and their protective role in hippocampal neuron apoptosis induced by oxygen and glucose deprivation

C-Jun N-terminal kinase 3 (JNK3) activation plays an essential role in the pathophysiology of cerebral ischemia. However, to date, no specific interventions with good efficacy have been reported. Therefore, in this study, we constructed a PLGA/JNK3-shRNA nanoparticle and examined its effects on neuronal apoptosis in an in vitro model of cerebral ischemia (oxygen and glucose deprivation model, OGD model). Herein, three JNK3-specific siRNAs were designed and synthesized, and their effects on JNK mRNA transcription were investigated; the most efficacious JNK3-specific siRNA was selected for recombination of the GV107/JNK3-shRNA plasmid. The PLGA/JNK3-shRNA nanoparticle was constructed, and its surface characterizations were confirmed. The roles of PLGA/JNK3-shRNA in neuronal JNK3 mRNA transcription, protein expression and activation as well as cell apoptosis were examined in a rat hippocampal neuron OGD model and compared with those of Lipofectamine 2000-mediated JNK3-siRNA transfection. The recombinant plasmid GV107/JNK3-shRNA was successfully constructed using siRNA1928. The PLGA/JNK3-shRNA nanoparticles were prepared as a sphere with a complete shape and smooth surface. The particle was about 225.4 nm in diameter with an average drug loading of 36.9%. OGD can cause marked cell apoptosis, whereas PLGA/JNK3-shRNA exposure can partly inhibit apoptosis. Further analysis demonstrated that the levels of JNK3 mRNA and protein as well as their activation were suppressed by PLGA/JNK3-shRNA nanoparticles. Compared with JNK3-siRNA delivered by Lipofectamine-2000, PLGA/JNK3-shRNA nanoparticles induced more JNK3 mRNA and protein reduction and more anti-apoptotic effects. To conclude, the PLGA/JNK3-shRNA nanoparticles could achieve good effects on inhibiting JNK3 signaling and neuronal apoptosis, and their preparation was feasible.