论文部分内容阅读
目的凋亡或程序性细胞死亡在多细胞生物体中已经被广泛研究,然而,关于单细胞寄生性原生动物细胞凋亡发生的分子机制却知之甚少。本研究旨在了解甲硝唑诱导阴道毛滴虫细胞凋亡的特征。方法培养阴道毛滴虫并用不同浓度的甲硝唑进行处理。在不同的时间间隔进行活细胞计数。提取甲硝唑处理过的阴道毛滴虫基因组进行DNA断裂片段检测。用DNA断端末端标记(TUNEL)法测定甲硝唑处理后阴道毛滴虫核酸内切酶活性。流式细胞检测分析脂酰丝氨酸暴露情况。结果甲硝唑可以诱导阴道毛滴虫出现凋亡样细胞死亡。这种凋亡样细胞死亡表现为细胞皱缩,磷脂酰丝氨酸暴露以及核染色体凝聚,但并未检测到寡核苷酸DNA梯带。结论阴道毛滴虫程序性细胞死亡的调节通路不同于多细胞生物体。确定导致原生动物细胞死亡的凋亡通路也许最终可用于鉴定新的治疗靶点。
Apoptosis or programmed cell death has been widely studied in multicellular organisms, however, little is known about the molecular mechanisms involved in the apoptosis of unicellular parasitic protozoa. This study aimed to understand the characteristics of metronidazole-induced apoptosis in Trichomonas vaginalis cells. Methods Trichomonas vaginalis was cultured and treated with different concentrations of metronidazole. Live cell counts are taken at different time intervals. The metronidazole-treated Trichomonas vaginalis genome was extracted for DNA fragmentation detection. The endotoxin activity of Trichomonas vaginalis after metronidazole treatment was determined by TUNEL method. Flow cytometry analysis of lipid acyl serine exposure. Results Metronidazole can induce apoptosis of Trichomonas vaginalis-like cells. This apoptosis-like cell death was characterized by cellular shrinkage, phosphatidylserine exposure, and nuclear chromosomal aggregation, but no detectable oligonucleotide DNA ladder. Conclusion Trichomonas vaginalis programmed cell death pathway is different from that of multicellular organisms. Determining the apoptotic pathway leading to protozoan cell death may eventually be used to identify new therapeutic targets.