Experimental study of scoring model for liver cirrhosis diseases

Abstract

Objective To investigate the effect of Toxoplasma gondii ME49 strain on the apoptosis of mouse placental trophoblastic cells in vitro.Methods Mouse placental trophoblastic cells (concentration of 5 × 106 /ml) were cultured in the different cell culture vessels.The cells were treated for 8 h with different concentrations of Toxoplasma gondii ME49 strain (the concentration of tachyzoites was 2 × 106 /ml,4 × 106 /ml,and 8 × 106 /ml,respectively).FCM was used to examine the apoptosis rates of the placental trophoblastic cells stained with the fluorescent dye of Annexin Ⅴ-FITC/PI;fluorescence microscopy was used to observe the changes of cellular morphology,and Western blotting analysis was used to detect the protein levels of Bax and Bcl-2.Results The trophoblastic cells infected with Toxoplasma gondii ME49 strain showed a higher apoptosis compared to the normal cells(P 0.05),and the apoptosis rates increased with the concentration of tachyzoites in the infected groups.The highest apoptosis rate was 28.37% which was found 8 h after culture with 8 ×106 /ml tachyzoites.Fluorescence microscope observed that the apoptosis of trophoblastic cells increased with the increase of Toxoplasma gondii.Western blotting analysis showed that the relative expression levels of Bax and Bcl-2 were 1.24 ±0.05,1.37 ± 0.03,1.78 ± 0.04,and 1.15 ± 0.03,1.09 ± 0.05,0.97 ± 0.01,respectively,which were significantly different from those of the control group (1.17 ±0.06,1.23 ± 0.02,P 0.05).Conclusion Infection with Toxoplasma gondii ME49 strain can promote the apoptosis of mouse placental trophoblastic cells in vitro through up-regulating Bax expression and down-regulating Bcl-2 expression.