刚地弓形虫培养上清对THP-1增殖的影响及可能机制

 利用弓形虫速殖子细胞内寄生的特性和对宿主细胞生物学行为的影响，提取弓形虫体外细胞共培养上清，并研究上清对人急性单核细胞白血病细胞THP-1增殖及细胞周期的影响。方法 取对数生长期的THP-1细胞（浓度为5×105）分别接种于不同细胞培养瓶，对照组加入含10%胎牛血清RPMI-1640，试验组加入相同体积不同数量(2×107mL-1、 4×107mL-1、 8×107 mL-1 )弓形虫速殖子培养上清孵育不同时间后， MTT法检测THP-1细胞增殖抑制率；流式细胞仪检测细胞周期改变；以Western印迹方法分析上清作用48h后细胞核转录因子NF-κB/p65与周期蛋白cyclinD1表达或活性的变化。结果 MTT法检测结果显示弓形虫培养上清抑制THP-1细胞株增殖，且呈时间剂量依赖性。流式细胞仪检测显示处理组细胞周期在G0/G1期产生阻滞； Western印迹方法分析THP-1细胞株的NF-κB/p65、cyclin D1 蛋白表达量下降,结论 刚地弓形虫培养上清能够抑制人急性单核细胞白血病细胞THP-1细胞株增殖并可通过细胞株的NF-κB信号途径来下调cyclin D1蛋白表达引起人急性单核细胞白血病细胞THP-1细胞株G0/G1期阻滞。

Effect of Culture Supernatant of Toxoplasma gondii on Human acute monocytic leukemia cell line THP-1 and probable mechanisms

The study on how to get culture supernatants of Toxoplasma gondii and investigate the inhibitive effects of culture supernatants of Toxoplasma gondii on the proliferation and on the cell cycle of Human acute monocytic leukemia cell line THP-1 in vitro. Methods THP-1 cells (concentration of 5×104mL-1) were harvested in the different cell culture plates. The cells were treated for different hours with different concentrations of Toxoplasma gondii culture supernatant. Growth inhibition rates were tested with the MTT method；Cell cycle were checked by Flow Cytometer. Western blot was used to detect the leves of NF-κB/p65 and cyclin D1 of cells. Results the culture supernatants of Toxoplasma gondii could inhibit the proliferation of THP-1 cells in a time-dose dependent manner. Cell cycle was significantly stopped at G0/G1phase by the culture supernatants with FCM technology. The culture supernatant of Toxoplasma gondii reduced the expressions of gene NF-κB 、cyclin D1 of THP-1 cells. Conclusion The culture supernatant of Toxoplasma gondii may inhibit THP-1 cell and arrest the cell cycle of THP-1 cells at G0/G1phase mainly by regulating the expression of gene NF-κB 、cyclin D1 .