Effects of Vibrio vulnificus cytolysin membrane pore-forming peptide on lysosome formation and ROS production in macrophages and its cytotoxicity
10.3760/cma.j.cn112309-20200303-00093
- VernacularTitle:创伤弧菌溶细胞素膜成孔肽对巨噬细胞溶酶体和ROS水平及细胞活性的影响
- Author:
Yi ZHENG
1
;
Xianhui HUANG
;
Danli XIE
;
Yongliang LOU
Author Information
1. 温州医科大学检验医学院，生命科学学院 325035
- From:
Chinese Journal of Microbiology and Immunology
2020;40(11):852-856
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the cytotoxicity of a recombinant Vibrio vulnificus cytolysin membrane pore-forming peptide (rMpf) to J774A.1 cells as well as its influences on reactive oxygen species (ROS) and lysosomes, and to analyze the cellular localization of the rMpf. Methods:The rMpf was expressed using an Escherichia coli expression system and then used to treat murine J774A.1 cells in vitro. The viability of rMpf- and PBS-treated J774A.1 cells and the levels of ROS and lysosomes in these cells were analyzed by flow cytometry. Confocal microscopy was used to observe the cellular localization of rMpf in the J774A.1 cells. Results:No significant differences in cell viability, ROS production or lysosome formation in J774A.1 cells were found between low-dose (4 μg/ml) rMpf-treated and untreated groups. However, the cell viability, ROS production and lysosome formation were significantly decreased after treating J774A.1 cells with higher doses of rMpf (20 and 60 μg/ml). Moreover, the rMpf was found to localize in both the cytoplasm and nuclei of J774A.1 cells in a dose-dependent manner.Conclusions:The rMpf could localize in both the cytoplasm and nuclei of J774A.1 cells. It would inhibit the cellular ROS production and lysosome formation to damage macrophage function. This study provided a novel sight for understanding the cytotoxic domain and pathogenesis of Vibrio vulnificus cytolysin and other membrane pore-forming cytolysins.
