ABSTRACT
BACKGROUND: Bacterial pathogens have many strategies for infecting and persisting in host cells. Adhesion, invasion and intracellular life are important features in the biology of mollicutes. The intracellular location ofUreaplasma diversum may trigger disturbances in the host cell. This includes activation or inhibition of pro and anti-apoptotic factors, which facilitate the development of host damage. The aim of the present study was to associate U. diversum infection in HEp-2 cells and apoptosis induction. Cells were infected for 72hs with four U. diversum clinical isolates and an ATCC strain. The U. diversuminvasion was analyzed by Confocal Laser Scanning Microscopy and gentamicin invasion assay. The apoptosis was evaluated using pro-apoptotic and anti-apoptotic gene expression, and FITC Annexin V/Dead Cell Apoptosis Kit. RESULTS: The number of internalized ureaplasma in HEp-2 cells increased significantly throughout the infection. The flow cytometry analysis with fluorochromes to detect membrane depolarization and gene expression for caspase 2, 3 and 9 increased in infected cells after 24 hours. However, after 72 hours a considerable decrease of apoptotic cells was observed. CONCLUSIONS: The data suggests that apoptosis may be initially induced by some isolates in association with HEp-2 cells, but over time, there was no evidence of apoptosis in the presence of ureaplasma and HEp-2 cells. The initial increase and then decrease in apoptosis could be related to bacterial pathogen-associated molecular pattern (PAMPS). Moreover, the isolates of U. diversum presented differences in the studied parameters for apoptosis. It was also observed that the amount of microorganisms was not proportional to the induction of apoptosis in HEp-2 cells.
Subject(s)
Humans , Female , Ureaplasma/pathogenicity , Ureaplasma Infections/physiopathology , Apoptosis/physiology , Time Factors , Ureaplasma/drug effects , Bacterial Adhesion , Actin Cytoskeleton/ultrastructure , Gentamicins/pharmacology , HeLa Cells/microbiology , Gene Expression , Cell Survival , Tumor Necrosis Factor-alpha/metabolism , Statistics, Nonparametric , Microscopy, Confocal , Caspase 3/metabolism , Caspase 2/metabolism , Caspase 9/metabolism , Real-Time Polymerase Chain Reaction , Flow Cytometry , Pathogen-Associated Molecular Pattern Molecules/metabolismABSTRACT
30 infertile patients were included in this study according to certain criteria, which are harboring Mycoplasma or Ureaplasma and being free from any other systemic or local factor that might influence semen. Another 30 patients whose semen samples were free from these organisms were included as control. Complete semen analysis, cytology and culture were done and data obtained were analyzed statistically. The study showed that Ureaplasma urealyticum has a significant effect on the following semen parameters: pH and liquefaction [increased], sperm motility [depressed], sperm concentration and viability [decreased] with increased percentage of abnormal forms. On the other hand, mycoplasma showed a significant effect only on the increased production of abnormal forms. These results point to the necessity for culture of these organisms, especially in the long standing idiopathic asthenozoosperm and/or increased percentage of abnormal forms in any barren marriage