中国动物传染病学报2024，Vol.32Issue(2)：27-34,8.
通过抑制caspase-8活性提高PRRSV体外培养滴度的初步研究
A Preliminary Research on Increasing the Titer of PRRSV in vitro Culture by Inhibiting the Activity of Caspase-8
摘要
Abstract
The purpose of the present study was to investigate the feasibility of promoting PRRSV replication by blocking apoptosis.Marc-145 cells were infected with a highly pathogenic porcine reproductive and respiratory syndrome virus(HP-PRRSV)strain and three apoptosis inhibitors(Z-VAD-FMK,Z-IETD-FMK and Z-LEHD-FMK)were used to respectively treat PRRSV-infected cells at different concentrations and times.At the same time,caspase-8 activity kit,quantificational real-time polymerase chain reaction(qRT-PCR)and flow cytometry were used to detect the expression of caspase-8 activity,apoptosis and pathway related factors under the best inhibitor.The results showed that z-IETD-FMK(caspases-8 specific inhibitor)was the optimal inhibitor at 10 μmol/L.Addition of 10 μmol/L Z-IETD-FMK at 48 h of PRRSV infection maximally inhibited the PRRSV titer.In addition,the caspase-8 activity of Marc-145 cells was significantly increased after PRRSV infection and,and extension of infection time caused both the apoptosis rate and proapoptotic factor Bax increased while the antiapoptotic factor Bcl-2 level decreased.After treatment with inhibitor z-IETD-FMK,the activity of caspases-8 went down and the apoptosis rate also decreased.On the other hand,the expression level of Bax was significantly lower than that in the viral infection group without inhibitor(P＜0.01).In contrast,the expression level of Bcl-2 increased,which was significantly higher than that in the viral infection group without inhibitor(P＜0.01).In conclusion,HP-PRRSV infection induced apoptosis of Marc-145 cells through caspase-8 pathway,preventing viral replication.Caspase-8 specific inhibitor effectively inhibited cell apoptosis thereby increasing the PRRSV titer.关键词
HP-PRRSV/caspase/细胞凋亡/活性检测Key words
Highly pathogenic porcine reproductive and respiratory syndrome virus/caspase/apoptosis/activity detection分类
畜牧业引用本文复制引用
钟华,白安斌,刘金凤,陈冰,覃绍敏,陈凤莲,马玲,林俊,毛燕,秦树英..通过抑制caspase-8活性提高PRRSV体外培养滴度的初步研究[J].中国动物传染病学报,2024,32(2):27-34,8.基金项目
广西生物技术兽医生物技术重点实验室开发项目(19-50-40-B-03) （19-50-40-B-03）
国家现代农业产业技术体系广西创新团队生猪产业功能专家岗位项目(nycytxgxcxtd--15-01) （nycytxgxcxtd--15-01）
