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OBJECTIVE To investigate the effect of Trichomonas vaginalis macrophage migration inhibitory factor (TvMIF) on THP-1 macrophages. METHODS Recombinant TvMIF protein was prokaryotic expressed and purified, and endotoxin was removed after identification. Following exposure to TvMIF at concentrations of 0, 1, 5, 10, 50 and 100 ng/mL, the cytotoxicity of the recombinant TvMIF protein to THP-1 macrophages was tested using cell counting kit (CCK)-8 assay, and the apoptosis of THP-1 macrophages and reactive oxygen species (ROS) were detected using flow cytometry. The relative expression of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), caspase-1, interleukin-1β (IL-1β) and IL-18 genes was quantified using real-time fluorescent quantitative PCR (qPCR) assay, and the expression of caspase-1, NLRP3, gasdermin D (GSDMD), gasdermin D N-terminal (GSDMD-NT) and pro-IL-1β proteins were determined using Western blotting assay. RESULTS Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) displayed successful expression and purification of the recombinant TvMIF protein with a molecular weight of 15.5 kDa, and the endotoxin activity assay showed the successful removal of endotoxin in the recombinant TvMIF protein (endotoxin concentration < 0.1 EU/mL), which was feasible for the subsequent studies on protein functions. Flow cytometry revealed that the recombinant TvMIF protein at a concentration of 10 ng/mL and less promoted the apoptosis of THP-1 macrophages, and the highest apoptotic rate of THP-1 macrophages was seen following exposure to the recombinant TvMIF protein at a concentration of 5 ng/mL, while the recombinant TvMIF protein at concentrations of 50 and100 ng/mL inhibited the apoptosis of THP-1 macrophages. Exposure to the recombinant TvMIF protein at a concentration 1 ng/mL resulted in increased ROS levels in THP-1 macrophages. qPCR assay quantified significantly elevated caspase-1, NLRP3, IL-18 and IL-1β expression in THP-1 macrophages 8 hours post-treatment with the recombinant TvMIF protein at a concentration 1 ng/mL, and Western blotting determined increased caspase-1, NLRP3, pro-IL-1β, GSDMD and GSDMD-NT protein expression in THP-1 macrophages following exposure to the recombinant TvMIF protein at a concentration 1 ng/mL. Pretreatment with MCC950 significantly reduced GSDMD and GSDMD-NT protein expression. CONCLUSIONS High-concentration recombinant TvMIF protein inhibits macrophage apoptosis, while low-concentration recombinant TvMIF protein activates NLRP3 inflammasome and promotes macrophage pyroptosis.
L Lin, and
S Huang, and
F Zhang, and
J Li, and
X Jiang, and
S Chen
June 2018,
mBio,
L Lin, and
S Huang, and
F Zhang, and
J Li, and
X Jiang, and
S Chen
June 1974,
Zhonghua Minguo wei sheng wu xue za zhi = Chinese journal of microbiology,
L Lin, and
S Huang, and
F Zhang, and
J Li, and
X Jiang, and
S Chen
June 2014,
Proceedings of the National Academy of Sciences of the United States of America,
L Lin, and
S Huang, and
F Zhang, and
J Li, and
X Jiang, and
S Chen
February 1980,
Indian journal of experimental biology,
L Lin, and
S Huang, and
F Zhang, and
J Li, and
X Jiang, and
S Chen
June 2022,
Andrologia,
L Lin, and
S Huang, and
F Zhang, and
J Li, and
X Jiang, and
S Chen
January 1999,
BioFactors (Oxford, England),
L Lin, and
S Huang, and
F Zhang, and
J Li, and
X Jiang, and
S Chen
January 2019,
Parasitology research,
L Lin, and
S Huang, and
F Zhang, and
J Li, and
X Jiang, and
S Chen
January 1958,
Comptes rendus des seances de la Societe de biologie et de ses filiales,
L Lin, and
S Huang, and
F Zhang, and
J Li, and
X Jiang, and
S Chen
June 1971,
Japanese journal of medical science & biology,
L Lin, and
S Huang, and
F Zhang, and
J Li, and
X Jiang, and
S Chen
January 2002,
Critical care medicine,
