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Anti-inflammatory effects of higenamine (Hig) on LPS-activated mouse microglia (BV2) through NF-κB and Nrf2/HO-1
Songwei Yang1, Shifeng Chu2, Qidi Ai3
1Hunan University of Chinese Medicine & Hunan Engineering Technology Center of Standardization and Function of Chinese Herbal Decoction Pieces, Changsha 410208, Hunan, China; State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia & Neuroscience Center, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.
Abstract:
Microglia are the most widely equipped protective cells in the brain and play a pivotal role in the development of neurological diseases. Inflammatory response and oxidative stress are critical risk factors in the activation of microglia which may cause various neurological diseases. Higenamine (Hig), a plant-based alkaloid and isolated from Aconite tuber, exhibits various properties and is mainly applied to treat heart failure. In addition, Hig expresses potential protective effects for neurodegenerative diseases. However, the effects and mechanisms of Hig on lipopolysaccharide (LPS) activated mouse microglia has not been fully explored. Therefore, we evaluated the anti-inflammatory effects of Hig on LPS-activated BV2 microglia and revealed the underlying mechanisms. Our data showed that Hig significantly inhibited the production of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), reactive oxygen species (ROS) as well as NO (mediated by iNOS) and PGE2 (mediated by COX2) in LPS-activated BV2 cells. Then we found that Hig suppressed NF-κB signaling pathway by inhibiting nuclear translocation of NF-κB/p65 subunit as well as degradation and phosphorylation of IκBα in cytoplasm, and the effect of Hig was intimately related to NF-κB inhibitor BAY-11-7082. Furthermore, we found that the anti-inflammatory effect of Hig were accompanied by the promotion of heme oxygenase-1 (HO-1) and nuclear factor erythroid 2-related factor-2 (Nrf2) expression, which was partly reversed by protoporphyrin (SnPP) and Nrf2 siRNA, respectively. Taken together, our results demonstrated that Hig expressed significant anti -inflammatory and -oxidative effects by inhibiting NF-κB and activating Nrf2/HO-1 signaling pathways.
Insights
Higenamine (Hig) reduces inflammation and oxidative stress in activated microglia by inhibiting the NF-κB pathway and activating the Nrf2/HO-1 pathway, offering potential neuroprotection.
Area of Science:
- Neuroscience
- Pharmacology
- Cell Biology
Background:
- Microglia are crucial brain immune cells implicated in neurological diseases.
- Microglial activation by inflammatory stimuli like lipopolysaccharide (LPS) drives neuroinflammation and oxidative stress.
- Higenamine (Hig), a plant alkaloid, shows potential neuroprotective effects, but its mechanisms in activated microglia are unclear.
Purpose of the Study:
- To investigate the anti-inflammatory and antioxidant effects of Higenamine (Hig) on lipopolysaccharide (LPS)-activated BV2 microglia.
- To elucidate the underlying molecular mechanisms, focusing on the NF-κB and Nrf2/HO-1 signaling pathways.
Main Methods:
- BV2 microglia were activated with LPS, and then treated with Higenamine (Hig).
- Key inflammatory mediators (TNF-α, IL-6, NO, PGE2) and reactive oxygen species (ROS) were quantified.
- NF-κB pathway activation (p65 translocation, IκBα degradation/phosphorylation) and Nrf2/HO-1 pathway activation were assessed.
- Specific inhibitors (BAY-11-7082, SnPP) and gene silencing (Nrf2 siRNA) were used to confirm mechanisms.
Main Results:
- Hig significantly reduced LPS-induced production of TNF-α, IL-6, NO, PGE2, and ROS in BV2 cells.
- Hig suppressed the NF-κB signaling pathway by inhibiting IκBα degradation and p65 nuclear translocation.
- Hig promoted the expression of heme oxygenase-1 (HO-1) and nuclear factor erythroid 2-related factor-2 (Nrf2).
- These effects were partly dependent on Nrf2 activation and mediated via NF-κB inhibition.
Conclusions:
- Higenamine (Hig) exerts significant anti-inflammatory and antioxidant effects on LPS-activated microglia.
- The mechanisms involve the inhibition of the NF-κB signaling pathway and the activation of the Nrf2/HO-1 pathway.
- Hig demonstrates therapeutic potential for neurological diseases associated with microglial activation.
