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| 刚地弓形虫ME49株对体外培养的小鼠胎盘滋养细胞凋亡的影响 |
| 葛璞1,2,车艺1,2,林玲1,2,黄开顺3,叶彬2,4* |
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| (1.重庆医科大学病理生理学教研室,重庆 400016 ;2.重庆医科大学分子医学与肿瘤中心,重庆 400016 ;3.重庆医科大学生命科学院,重庆 400016;4.重庆医科大学病原生物学教研室,重庆 400016) |
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| 摘要: |
| 目的 研究刚地弓形虫ME49株对体外培养的孕期小鼠胎盘滋养层细胞凋亡的影响。方法 制备小鼠胎盘滋养层细胞并分别接种于不同细胞培养板,对照组加入DMEM高糖培养液孵育,实验组加入相同体积不同密度(2×106 /ml、4×106 /ml、8×106/ml )弓形虫速殖子培养8 h后,Annexin Ⅴ-FITC/PI染色细胞后上流式细胞仪检测各个时间点细胞凋亡率变化,以蛋白质印迹法分析凋亡相关蛋白Bax、Bcl-2的表达。结果 流式细胞仪检测弓形虫感染后的滋养层细胞凋亡率较对照组有升高趋势(P<0.05),呈量效依赖性,弓形虫密度8×106/ml组8 h凋亡率最高,为28.37%；荧光显微镜观察,随弓形虫感染密度增加,滋养层细胞凋亡数目增多。蛋白质印迹法检测刚地弓形虫ME49株作用于滋养层细胞8 h后实验组的Bax、Bcl-2蛋白表达(与β-actin比值)各为1.24±0.05、1.37±0.03、1.78±0.04与1.15±0.03、1.09±0.05、0.97±0.01,较对照组的比值(1.17±0.06和1.23±0.02)有明显的改变(P<0.05)。结论 刚地弓形虫ME49株可促进体外培养的孕期小鼠滋养细胞正常凋亡,其机制可能与滋养细胞Bax表达上调和Bcl-2表达下调有关。 |
| 关键词: 刚地弓形虫ME49株 寄生虫性妊娠并发症 滋养细胞 细胞凋亡 |
| DOI：10.3724/SP.J.1008.2010.0485 |
| 投稿时间：2010-01-15修订日期：2010-04-23 |
| 基金项目: |
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| Effect of Toxoplasma gondii ME49 strain on apoptosis of mouse placental trophoblastic cells |
| GE Pu1,2, CHE Yi1,2, LIN Ling1,2 , HUANG Kai-shun3, YE Bin2,4* |
| (1. Department of Pathophysiology, Chongqing Medical University, Chongqing 400016, China;2. Center for Molecular Medical and Tumor Research, Chongqing Medical University, Chongqing 400016, China;3. Department of Life Science, Chongqing Medical University, Chongqing 400016, China;4. Department of Pathobiology, Chongqing Medical University, Chongqing 400016, China) |
| 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. |
| Key words: Toxoplasma gondii ME49 strain parasitic pregnancy complications trophoblastic cells apoptosis |
