摘要
摘要：目的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增殖及凋亡的影响。方法收集对数生长期的THP-1细胞以5X10^7／ml细胞浓度接种于不同培养瓶中，对照组加入含10％胎牛血清的RPMll640，实验组加入相同体积不同数量（2×10^7／ml、4X10^7／ml、8×10^7／m1）弓形虫速殖子培养上清，采用四甲基氮噻唑蓝（MTY）法检测吸光度（A490值）并计算THP-1细胞增殖抑制率；倒置显微镜下观察细胞形态变化；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Western印迹方法分析凋亡相关蛋白Bax、Bcl-2的表达或活性。结果MTY法检测结果弓形虫培养上清呈时间剂量依赖性抑制THP-1细胞株增殖，倒置显微镜下观察处理组细胞有发泡现象和凋亡小体出现。流式细胞仪检测弓形虫感染后的THP-1细胞凋亡率较对照组有升高趋势（P〈0．05），呈量效依赖性，Westernblot检测刚地弓形虫培养上清作用于THP-l细胞48h后实验组的Bax、Bcl-2蛋白表达较对照组的比值分别有明显的升高与降低（P〈0．05）。结论刚地弓形虫速殖子培养上清对体外培养THP-l细胞增殖有明显的抑制作用，并可诱导THP-1细胞凋亡。
Objective The study on the inhibitive effects of culture supernatants of Toxoplasma gondii on the proliferation of Human acute monocytic leukemia cell line THP-1 in vitro. Methods THP-1 cells were treated with different concentrations of culture supernatants of Toxo- plasma gondii （ the concentration of tachyzoites was 2 ×107/ml, 4×107/ml, 8×107/ml respectively）. MTY method was tested the growth inhibition rates ; Invert microscopy was used to observe the changes of cellular morphology ; FCM was used to test the rates of ap0ptosis of the THP-1 cells, Western blot was used to detect the protein expressions of Bax, Bcl-2 of cells. Results The culture supernatant of Toxoplasma gondii inhibited the proliferation of THP-I ceils. The classic cellular morphology of apoptosis in the treated THP-1 cells was observed under the invert microscope. The protein expressions of Bcl-2/Bax were observed by Western blot. Conclusion the culture supernatant of Toxoplasma zondii revealed an inhibition to the proliferation of THP-1 cells and induced their apoptosis in vitro.
出处
《世界科技研究与发展》
CSCD
2011年第6期1066-1069,共4页
World Sci-Tech R&D
基金
国家自然科学基金(X9100-0200040502)资助项目
关键词
刚地弓形虫培养上清
人急性单核细胞白血病细胞THP-1
增殖
凋亡
：culture supernatants of Toxoplasma gondii
Human acute monocytic leukemia cell line TI-IP-1
proliferation
apoptosis
