Study on programmed cell death caused by severe fever with thrombocytopenia syndrome virus
10.3760/cma.j.cn112866-20220923-00199
- VernacularTitle:发热伴血小板减少综合征病毒致细胞程序性死亡的研究
- Author:
Zhu YANG
1
;
Chang LIU
;
Zeming QIN
;
Hongling WEN
;
Tao HUANG
;
Li ZHAO
Author Information
1. 山东大学公共卫生学院卫生微生物检验学系 山东省"十三五"高校重点实验室，济南 250012
- Keywords:
Severe fever with thrombocytopenia syndrome virus;
Programmed cell death;
Pyroptosis
- From:
Chinese Journal of Experimental and Clinical Virology
2023;37(1):1-6
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the form of programmed cell death (PCD) induced by severe fever with thrombocytopenia syndrome virus (SFTSV) infection in Vero cells and further explore the existence of pyroptosis, so as to provide new ideas for studying the pathogenic mechanism of SFTSV.Methods:Vero cells were infected with SFTSV at different multiplicity of infection (MOI), cytopathic effect (CPE) was observed daily, cell viability was detected by CCK-8 method, and cell membrane damage was detected by LDH release test to determine the optimal amount of virus infection and cell death time. Vero cells were pretreated with different PCD inhibitors and infected with SFTSV. CCK-8 kit was used to detect the cell viability and determine the death form of PCD caused by SFTSV. The expression of pyroptosis related proteins was detected by Western blotting to further explore the existence of pyroptosis.Results:At 48 h and 72 h after SFTSV infected Vero cells with MOI=10, the optimal infection amount and time of subsequent experiments were observed. At this time, the CPE of cells was obvious, the cell viability decreased to 51% and 41% of the control group ( P<0.001, P<0.001), and the LDH release amount reached 24% and 37% of the maximum enzyme activity release wells, were 3.8, 3.4 times of LDH release in the control group ( P<0.001, P<0.001). The inhibition of SFTSV-induced cell death by different PCD inhibitors showed that pan-caspase inhibitor and receptor-interacting serine/threonine-protein kinase 3 (RIP3) inhibitor had inhibitory effects at 48 h and 72 h. The cell viability was 2.1 and 1.6 times of the viral control group at 48 h, 2.3 and 1.7 times of the viral control group at 72 h, and the effect of pan-caspase inhibitor was significantly higher than that of the other inhibitor groups. Caspase-1 and caspase-3 inhibitors only had inhibitory effect at 48 h, and the cell viability was 1.5 and 1.3 times of the viral control group. After SFTSV infection of Vero cells, the expressions of caspase-1 and IL-1β increased gradually with the prolongation of time, and reached 3.4 and 9.5 times of the control group at 48 h, respectively ( P<0.001, P<0.001). Conclusions:SFTSV infection of Vero cells can lead to various forms of PCD, including apoptosis, pyroptosis and programmed necrosis, and pyroptosis related protein activation can be detected in the process of PCD, which further explored the existence of pyroptosis.
