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Role of Militarine in PM2.5-Induced BV-2 Cell Damage
Shu-Xin Tian1, Wen Cheng1, Jing-Jing Lu1
1College of Medical Technology, Zhejiang Chinese Medical University, Hangzhou, 310053, China.
Abstract:
A growing number of studies have shown that air fine particulate matter (PM2.5) pollution is closely associated with neuroinflammation in humans. Militarine, a glucosyloxybenzyl 2-isobutylmalate compound isolated from Bletilla striata, has been found to exert significant neuroprotective effects. However, the anti-inflammatory, antioxidant and antiapoptotic effects of militarine on PM2.5-stimulated BV-2 microglial cells have not been reported. This study aimed to investigate the protective effects of militarine against PM2.5-induced cytotoxicity and its mechanism in BV-2 microglial cells. Our results revealed that pretreatment with 0.31-1.25 μg/mL militarine reversed the morphological changes caused by PM2.5 and decreased proinflammatory cytokine generation and gene expression in PM2.5-treated BV-2 cells. In particular, tumor necrosis factor-α and interleukin-6 expression was inhibited in a dose-dependent manner. Notably, militarine markedly inhibited the upregulation of Toll-like receptor 4, Toll-like receptor 2, and cyclo-oxygenase-2 expression at both the mRNA and protein levels and reduced NF-κB pathway-associated protein expression. Immunofluorescence analysis showed that militarine suppressed NF-κB activity through inhibiting p65 nuclear translocation. Our data suggested that militarine alleviated neuroinflammation in BV-2 microglial cells, possibly by inhibiting the expression of neuroinflammatory cytokines through the TLR/NF-κB signaling pathway. Additionally, militarine significantly reduced PM2.5-mediated reactive oxygen species (ROS) generation and cell apoptosis and restored the mitochondrial membrane potential (MMP; ΔΨm). Collectively, these findings demonstrate that militarine played a protective role against PM2.5-induced damage in BV-2 cells by exerting anti-inflammatory, antioxidant, and antiapoptotic effects.
Insights
Militarine protects against air pollution-induced neuroinflammation by reducing inflammatory markers and oxidative stress in microglial cells. This compound demonstrates antioxidant and antiapoptotic effects, mitigating PM2.5 damage.
Area of Science:
- Neuroscience
- Environmental Health
- Pharmacology
Background:
- Air fine particulate matter (PM2.5) is linked to human neuroinflammation.
- Militarine, from Bletilla striata, shows neuroprotective potential.
- The effects of militarine on PM2.5-induced microglial cell damage are unknown.
Purpose of the Study:
- To investigate the protective effects of militarine against PM2.5-induced cytotoxicity in BV-2 microglial cells.
- To elucidate the underlying mechanisms of militarine's action, including its anti-inflammatory, antioxidant, and antiapoptotic properties.
Main Methods:
- BV-2 microglial cells were pretreated with varying concentrations of militarine before PM2.5 exposure.
- Assessed cell morphology, proinflammatory cytokine levels (TNF-α, IL-6), and gene/protein expression (TLR4, TLR2, COX-2, NF-κB pathway).
- Analyzed reactive oxygen species (ROS) generation, apoptosis, and mitochondrial membrane potential (MMP).
Main Results:
- Militarine reversed PM2.5-induced morphological changes and reduced proinflammatory cytokine expression.
- Militarine inhibited TLR/NF-κB signaling pathway activation, including p65 nuclear translocation.
- Militarine decreased ROS generation, reduced apoptosis, and restored MMP in PM2.5-treated cells.
Conclusions:
- Militarine exerts anti-inflammatory, antioxidant, and antiapoptotic effects against PM2.5-induced neurotoxicity in BV-2 cells.
- The protective mechanism involves the inhibition of the TLR/NF-κB signaling pathway.
- Militarine shows promise as a therapeutic agent against air pollution-related neuroinflammation.
