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[Knockdown of PRDX6 in microglia reduces neuron viability after OGD/R injury]
Li Tan1, Yong Zhao1, Beibei Jiang1
1Department of Pathology, Molecular Medicine and Cancer Research Center, College of Basic Medical Sciences, Chongqing Medical University, Chongqing 400016, China.
Abstract:
Objective To observe the effects of peroxiredoxin 6 (PRDX6) knockdown in the microglia on neuron viability after oxygen-glucose deprivation and reoxygenation (OGD/R). Methods Microglia was treated with lentivirus PRDX6-siRNA and Ca(2+)-independent phospholipase A2 (iPLA2) inhibitor, 1-hexadecyl-3-(trifluoroethgl)-sn-glycerol-2 phosphomethanol (MJ33). Twenty-four hours later, it was co-cultured with primary neuron to establish the microglia-neuron co-culture OGD/R model. According to the different treatment of microglia, the cells were divided into normal group, OGD/R group, negative control-siRNA treated OGD/R group, PRDX6-siRNA treated OGD/R group and PRDX6-siRNA combined with MJ33 treated OGD/R group. Western blot analysis and real-time quantitative PCR were respectively performed to detect PRDX6 protein and mRNA levels after knockdown of PRDX6 in microglia. The iPLA2 activity was measured by ELISA. MTS and lactate dehydrogenase (LDH) assay were used to measure neuron viability and cell damage. The oxidative stress level of neuron was determined by measuring superoxide dismutase (SOD) and malonaldehyde (MDA) content. Results In PRDX6-siRNA group, neuron viability was inhibited and oxidative stress damage was aggravated compared with OGD/R group. In PRDX6-siRNA combined with MJ33 group, cell viability was promoted and oxidative stress damage was alleviated compared with PRDX6-siRNA group. Conclusion PRDX6 in microglia protects neuron against OGD/R-induced injury, and iPLA2 activity has an effect on PRDX6.
Insights
Peroxiredoxin 6 (PRDX6) in microglia protects neurons from oxygen-glucose deprivation and reoxygenation (OGD/R) injury. Inhibiting PRDX6 worsens neuronal damage, while blocking Ca(2+)-independent phospholipase A2 (iPLA2) activity with MJ33 offers protection.
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- Microglia play a crucial role in neuroinflammation and neuronal injury.
- Oxygen-glucose deprivation and reoxygenation (OGD/R) models are used to study ischemic brain injury.
- Peroxiredoxin 6 (PRDX6) is an enzyme with antioxidant properties.
Purpose of the Study:
- To investigate the role of peroxiredoxin 6 (PRDX6) in microglia during oxygen-glucose deprivation and reoxygenation (OGD/R) induced neuronal injury.
- To determine the effect of PRDX6 knockdown in microglia on neuron viability.
- To explore the involvement of Ca(2+)-independent phospholipase A2 (iPLA2) activity in PRDX6-mediated neuroprotection.
Main Methods:
- Primary microglia and neurons were co-cultured to establish a microglia-neuron co-culture OGD/R model.
- Lentivirus-mediated PRDX6-siRNA was used to knockdown PRDX6 in microglia.
- The iPLA2 inhibitor MJ33 was administered to assess its effect on PRDX6 knockdown.
- Neuron viability was assessed using MTS and lactate dehydrogenase (LDH) assays.
- Oxidative stress markers, superoxide dismutase (SOD) and malonaldehyde (MDA), were measured.
Main Results:
- Knockdown of PRDX6 in microglia significantly inhibited neuron viability and aggravated oxidative stress damage following OGD/R.
- Treatment with the iPLA2 inhibitor MJ33 in combination with PRDX6-siRNA promoted cell viability and alleviated oxidative stress damage compared to PRDX6-siRNA alone.
- PRDX6 knockdown led to increased levels of oxidative stress markers (MDA) and decreased levels of antioxidant enzymes (SOD) in neurons.
Conclusions:
- Microglial PRDX6 plays a protective role against OGD/R-induced neuronal injury.
- iPLA2 activity is implicated in the protective mechanism of PRDX6 in microglia.
- Targeting microglial PRDX6 and modulating iPLA2 activity may represent therapeutic strategies for ischemic brain injury.
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