The P2X7 Receptor Involved in gp120-Induced Cell Injury in BV2 Microglia
Qiang Chen1, Hui Wu2, Shanshan Qin1
1Department of Physiology, Basic Medical College, Nanchang University, Nanchang, 330006, People's Republic of China.
Abstract:
This study was aimed at exploring the effects of P2X7 receptor on BV2 microglia cell injury induced by glycoprotein gp120 (gp120) and its underlying mechanisms. We used the MTS method to study the influence of different gp120 concentrations on BV2 microglia cells, and to test the degree of cell injury in each gp120 treatment group; quantitative real-time PCR (qPCR) and Western blot were used to detect the P2X7 mRNA and receptor protein expressions. Immunocytochemistry and Western blot were used to detect the P2X7 receptor expression and P65 NF-κB, respectively. We also measured the content of TNFα, IL-1β, nitric oxide (NO) and reactive oxygen species (ROS). We found that the cell survival rate generally decreased as gp120 concentration increased, and the cell survival rate of the gp120 + Brilliant Blue G (BBG) group was higher than that of the gp120 group. Western blot and qPCR results showed that the expressions of P2X7 receptor protein and mRNA were positively dose-dependent with gp120 concentration; the results of immunocytochemistry and Western blot showed that the expressions of P2X7 receptor and P65 NF-κB in the gp120 group increased significantly compared to those of the control (Ctrl) group, but those in the gp120+BBG group decreased. Taken together, these results confirmed that the P2X7 receptor is involved in gp120-induced BV2 microglial cell injury and that the underlying mechanism may be associated with the over-activation of microglia caused by P2X7 receptor up-regulation, which leads to abundant release of inflammatory factors which exert toxic effects on the cells.
Insights
The P2X7 receptor contributes to BV2 microglia cell injury caused by glycoprotein gp120. Blocking this receptor with Brilliant Blue G (BBG) reduces inflammation and cell damage, suggesting a therapeutic target.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Glycoprotein gp120 is implicated in neuroinflammation.
- Microglia play a crucial role in the central nervous system's immune response.
- The P2X7 receptor is a key mediator in inflammatory processes.
Purpose of the Study:
- To investigate the role of the P2X7 receptor in glycoprotein gp120-induced BV2 microglia cell injury.
- To elucidate the underlying mechanisms of P2X7 receptor involvement in this process.
Main Methods:
- BV2 microglia cell culture and treatment with varying concentrations of glycoprotein gp120 and Brilliant Blue G (BBG).
- MTS assay for cell viability assessment.
- Quantitative real-time PCR (qPCR) and Western blot for gene and protein expression analysis (P2X7 receptor, P65 NF-κB).
- Immunocytochemistry for P2X7 receptor localization.
- Measurement of inflammatory mediators: TNFα, IL-1β, nitric oxide (NO), and reactive oxygen species (ROS).
Main Results:
- Glycoprotein gp120 exposure led to a dose-dependent decrease in BV2 microglia cell survival.
- P2X7 receptor and P65 NF-κB expression levels were significantly upregulated by glycoprotein gp120.
- Treatment with BBG, a P2X7 receptor antagonist, attenuated glycoprotein gp120-induced cell injury and inflammatory mediator release.
- Increased P2X7 receptor expression correlated with elevated levels of TNFα, IL-1β, NO, and ROS.
Conclusions:
- The P2X7 receptor is critically involved in glycoprotein gp120-induced BV2 microglia cell injury.
- Upregulation of the P2X7 receptor leads to microglial over-activation and subsequent release of inflammatory factors, contributing to cell damage.
- Targeting the P2X7 receptor may represent a potential therapeutic strategy for neuroinflammatory conditions associated with glycoprotein gp120.
