Lv Yingjun, Bao Endong
College of Veterinary Medicine, Nanjing Agricultural University, Weigang 1, Nanjing 210095, China.
Toxicol In Vitro. 2009 Oct;23(7):1360-4. doi: 10.1016/j.tiv.2009.06.026. Epub 2009 Jun 27.
This study investigated apoptosis induced in chicken embryo fibroblast (CEF) cells by the toxic effects of polyinosinic:polycytidylic acid copolymer (Poly I:C). Along with cell viability and morphological changes, other indicators were measured in vitro after incubation of the cells with Poly I:C, including phosphatidylserine translocation, the formation of DNA fragments, activity of caspase-3, caspase-8 and caspase-9, and the expression levels of RIPK1 mRNA and TNRSF8 mRNA. An inhibition of proliferation was found with increasing levels of Poly I:C, showing that Poly I:C was toxic to the cells. The findings of phosphatidylserine translocation and formation of DNA fragmentation suggested that Poly I:C induced apoptosis. With increased incubation time, the activities of caspase-3 and caspase-8 increased, while there was no significant change in caspase-9 activity. Accordingly, it is concluded that the apoptosis induced by Poly I:C involves a cell death receptor-mediated pathway. The transcription level of RIPK1 mRNA decreased, while that of TNFRSF8 mRNA increased, indicating that Poly I:C-induced apoptosis was related to upregulation of TNFRSF8. These observations provide insight into the potential mechanism of Poly I:C-induced toxicity.
聚胞苷酸共聚物(Poly I:C)的毒性作用诱导鸡胚成纤维细胞(CEF)发生凋亡的情况。除细胞活力和形态变化外,在用Poly I:C孵育细胞后,还在体外测量了其他指标,包括磷脂酰丝氨酸易位、DNA片段形成、半胱天冬酶-3、半胱天冬酶-8和半胱天冬酶-9的活性,以及RIPK1 mRNA和TNRSF8 mRNA的表达水平。发现随着Poly I:C水平的升高,细胞增殖受到抑制,表明Poly I:C对细胞有毒性。磷脂酰丝氨酸易位和DNA片段化形成的结果表明Poly I:C诱导了凋亡。随着孵育时间的延长,半胱天冬酶-3和半胱天冬酶-8的活性增加,而半胱天冬酶-9的活性没有显著变化。因此,得出结论,Poly I:C诱导的凋亡涉及细胞死亡受体介导的途径。RIPK1 mRNA的转录水平降低,而TNFRSF8 mRNA的转录水平升高,表明Poly I:C诱导的凋亡与TNFRSF8的上调有关。这些观察结果为Poly I:C诱导毒性的潜在机制提供了见解。