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Methods to Study Lipid Alterations in Neutrophils and the Subsequent Formation of Neutrophil Extracellular Traps
Published on: March 29, 2017
Mitraphylline inhibits lipopolysaccharide-mediated activation of primary human neutrophils
Sergio Montserrat-de la Paz1, Angeles Fernandez-Arche2, Rocío de la Puerta2
1Department of Pharmacology, School of Pharmacy, University of Seville, 41012, Seville, Spain ; Laboratory of Cellular and Molecular Nutrition, Instituto de la Grasa, CSIC, Seville, Spain.
Background:
Mitraphylline (MTP) is the major pentacyclic oxindolic alkaloid presented in Uncaria tomentosa. It has traditionally been used to treat disorders including arthritis, heart disease, cancer, and other inflammatory diseases. However, the specific role of MTP is still not clear, with more comprehensivestudies, our understanding of this ancient herbal medicine will continue growing.
Hypothesis/Purpose:
Some studies provided its ability to inhibit proinflamatory cytokines, such as TNF-α, through NF-κB-dependent mechanism. TNF-α primes neutrophils and modulates phagocytic and oxidative burst activities in inflammatory processes. Since, neutrophils represent the most abundant pool of leukocytes in human blood and play a crucial role in inflammation, we aimed to determine the ability of MTP to modulate neutrophil activation and differentially regulate inflammatory-related cytokines.
Methods:
To determine the mechanism of action of MTP, we investigated the effects on LPS-activated human primary neutrophils responses including activation surface markers by FACS and the expression of inflammatory cytokines, measured by real time PCR and ELISA.
Results:
Treatment with MTP reduced the LPS-dependent activation effects. Activated neutrophils (CD16(+)CD62L(-)) diminished after MTP administration. Moreover, proinflamatory cytokines (TNF-α, IL-6 or IL-8) expression and secretion were concomitantly reduced, similar to basal control conditions.
Conclusion:
Taken together, our results demonstrate that MTP is able to elicit an anti-inflammatory response that modulates neutrophil activation contributing to the attenuation of inflammatory episodes. Further studies are need to characterize the mechanism by which MTP can affect this pathway that could provide a means to develop MTP as new candidate for inflammatory disease therapies.
Insights
Mitraphylline (MTP) reduces neutrophil activation and inflammatory cytokine release, showing potential as a novel therapy for inflammatory diseases. This study investigated MTP's anti-inflammatory effects on neutrophils.
Area of Science:
- Pharmacology
- Immunology
- Natural Products
Background:
- Mitraphylline (MTP) is a key alkaloid in Uncaria tomentosa, traditionally used for inflammatory conditions.
- Its precise anti-inflammatory mechanisms, particularly on neutrophils, require further elucidation.
Purpose of the Study:
- To investigate MTP's ability to modulate human neutrophil activation.
- To determine MTP's effect on inflammatory cytokine expression and secretion.
Main Methods:
- Human primary neutrophils were activated with LPS.
- Effects of MTP on neutrophil activation markers (FACS) and cytokine profiles (real-time PCR, ELISA) were analyzed.
Main Results:
- MTP treatment significantly reduced LPS-induced neutrophil activation (CD16(+)CD62L(-)).
- Expression and secretion of pro-inflammatory cytokines (TNF-α, IL-6, IL-8) were decreased by MTP.
Conclusions:
- MTP exhibits anti-inflammatory properties by modulating neutrophil activation and cytokine production.
- MTP represents a potential therapeutic candidate for inflammatory diseases.
