Author(s):
1. Mehreen Tariq:
Department of Biochemistry, Quaid-i-Azam University,Islamabad,Pakistan
2. Sehrish Gohar Ali:
Department of Biochemistry, Quaid-i-Azam University,Islamabad,Pakistan
3. Muhammad Rizwan Alam:
Department of Biochemistry, Quaid-i-Azam University,Islamabad,Pakistan
Abstract:
Neutrophil Extracellular Traps (NETs) are web-like filaments containing decondensed chromatin decorated with antimicrobial proteins that are released in response to inflammatory stimuli. Ionomycin and thapsigargin are potent inducers of NETosis (formation of NETs) which work via increase in intracellular calcium and ROS formation. The current study explored the role of mitochondrial calcium efflux in ionomycin and thapsigargininduced neutrophil degeneration that may be linked to cellular NETosis. Initially, Polymorphonuclear neutrophils (PMNs) in both whole blood (WB)and a platelet-lacking whole blood preparation (WBP) were pre-treated with CGP37157, a pharmacological blocker of NCLX, followed by stimulation with ionomycin and thapsigargin. Smears were Field's-stained and brightfield microscopy was performed for quantifying the morphological changes in neutrophils. Also, isolated PMNs treated with ionomycin and thapsigargin were used to check the release of MPO after stimulation. Moreover, mitochondrial membrane potential (MMP) was also assessed in ionomycin and thapsigargin treated PMNs. Our results showed increased neutrophil degeneration upon ionomycin and thapsigargin stimulation. The presence of platelets in whole blood negatively affected the ionomycin stimulation of neutrophils. Interestingly, CGP-37157 increased the damage in PMNs stimulated with ionomycin or thapsigargin. Inhibition of NCLX depolarized MMP in ionomycin stimulated neutrophils while thapsigargin stimulated neutrophils also induced depolarization of MMP. Our data also showed increased MPO activity in ionomycin and thapsigargin treated PMNs.
Page(s):
0-0
DOI:
DOI not available
Published:
Journal: First International Conference on Revamped Scientific Outlook of 21st Century (Abstract Book), Volume: 0, Issue: 0, Year: 2022
Keywords:
Mitochondrial Membrane potential
,
NETs
,
Thapsigargin
,
MPO Activity
,
Ionomycin
,
Neutrophil Degeneration
,
NCLX
References:
References are not available for this document.
Citations
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