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The attenuating effects of plumbagin on pro-inflammatory cytokine expression in LPS-activated BV-2 microglial cells
Samia S Messeha1, Najla O Zarmouh1, Patricia Mendonca1
1College of Pharmacy and Pharmaceutical Science, Florida A & M University, Tallahassee, FL 32307, USA.
Abstract:
Activated microglial cells produce the pro-inflammatory mediators such as nitric oxide (NO) and cytokines. The excessive release of these mediators can lead to neurodegenerative diseases, such as Alzheimer's disease (AD) and Parkinson's disease (PD). Inhibition of the release of these pro-inflammatory molecules may prevent or halt the progression of these diseases. Plumbagin (PL), a naphthoquinone compound in the roots of the traditional medicinal plant Plumbago zeylanica L., showed anti-inflammatory effects on macrophages. However, PL effects on activated microglia remain unknown. In the present study, PL has been examined for its anti-inflammatory effect on LPS - activated microglial BV-2 cells. In this study, NO and iNOS expression were investigated in BV-2 microglial cells in the presence of PL or the selective iNOS inhibitor L-N6-(1-iminoethyl) lysine (L-NIL). The results obtained indicate that PL was >30-fold potent than L-NIL in inhibiting NO production with an IC50 of 0.39μM. Our immunofluorescence study confirmed the ability of PL to significantly inhibit iNOS expression in the activated microglia. Furthermore, the extracellular microglial pro-inflammatory cytokine expression in the presence of 2μM of PL was detected, quantified, and validated using cytokine antibody protein arrays and quantitative ELISA. The results obtained showed that PL significantly downregulated the expression of many cytokines including IL-1α, G-CSF, IL-12 p40/p70, MCP-5, MCP-1, and IL-6. In conclusion, PL potency in attenuating multiple pro-inflammatory agents indicates its potential to be used for neurodegenerative diseases.
Insights
Plumbagin (PL), a natural compound, significantly reduces pro-inflammatory mediators in activated microglial cells. This suggests PL
Area of Science:
- Neuroscience
- Immunology
- Pharmacology
Background:
- Activated microglia release pro-inflammatory mediators like nitric oxide (NO) and cytokines.
- Excessive release of these mediators is implicated in neurodegenerative diseases such as Alzheimer's disease (AD) and Parkinson's disease (PD).
- Inhibiting these mediators may offer therapeutic potential for neurodegenerative conditions.
Purpose of the Study:
- To investigate the anti-inflammatory effects of Plumbagin (PL) on lipopolysaccharide (LPS)-activated microglial BV-2 cells.
- To evaluate PL's potency in inhibiting nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression.
- To assess PL's impact on the expression of extracellular pro-inflammatory cytokines in activated microglia.
Main Methods:
- Treatment of LPS-activated BV-2 microglial cells with varying concentrations of PL.
- Measurement of NO production and iNOS expression using immunofluorescence and IC50 determination.
- Quantification of extracellular pro-inflammatory cytokines using cytokine antibody protein arrays and quantitative ELISA.
Main Results:
- Plumbagin (PL) demonstrated over 30-fold greater potency than L-NIL in inhibiting NO production, with an IC50 of 0.39μM.
- PL significantly inhibited iNOS expression in activated microglia.
- PL treatment downregulated the expression of key pro-inflammatory cytokines, including IL-1α, G-CSF, IL-12 p40/p70, MCP-5, MCP-1, and IL-6.
Conclusions:
- Plumbagin (PL) exhibits significant anti-inflammatory properties by attenuating multiple pro-inflammatory mediators in activated microglia.
- The potent inhibition of NO, iNOS, and various cytokines by PL highlights its therapeutic potential for neurodegenerative diseases.
- Further research into PL as a treatment for conditions like Alzheimer's and Parkinson's disease is warranted.
