College of Coastal Agricultural Science of Guangdong Ocean University, Zhanjiang 524088, China.
College of Food Science and Technology of Guangdong Ocean University, Zhanjiang 524088, China.
J Vet Sci. 2024 Mar;25(2):e21. doi: 10.4142/jvs.23236.
Peste des petits ruminants (PPR) is a contagious and fatal disease of sheep and goats. PPR virus (PPRV) infection induces endoplasmic reticulum (ER) stress-mediated unfolded protein response (UPR). The activation of UPR signaling pathways and their impact on apoptosis and virus replication remains controversial.
To investigate the role of PPRV-induced ER stress and the IRE1-XBP1 and IRE1-JNK pathways and their impact on apoptosis and virus replication.
The cell viability and virus replication were assessed by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay, immunofluorescence assay, and Western blot. The expression of ER stress biomarker GRP78, IRE1, and its downstream molecules, PPRV-N protein, and apoptosis-related proteins was detected by Western blot and quantitative reverse transcription-polymerase chain reaction, respectively. 4-Phenylbutyric acid (4-PBA) and STF-083010 were respectively used to inhibit ER stress and IRE1 signaling pathway.
The expression of GRP78, IRE1α, p-IRE1α, XBP1s, JNK, p-JNK, caspase-3, caspase-9, Bax and PPRV-N were significantly up-regulated in PPRV-infected cells, the expression of Bcl-2 was significantly down-regulated. Due to 4-PBA treatment, the expression of GRP78, p-IRE1α, XBP1s, p-JNK, caspase-3, caspase-9, Bax, and PPRV-N were significantly down-regulated, the expression of Bcl-2 was significantly up-regulated. Moreover, in PPRV-infected cells, the expression of p-IRE1α, p-JNK, Bax, and PPRV-N was significantly decreased, and the expression of Bcl-2 was increased in the presence of STF-083010.
PPRV infection induces ER stress and IRE1 activation, resulting in apoptosis and enhancement of virus replication through IRE1-XBP1s and IRE1-JNK pathways.
小反刍兽疫（PPR）是一种传染性和致命性的绵羊和山羊疾病。PPRV 感染诱导内质网（ER）应激介导的未折叠蛋白反应（UPR）。UPR 信号通路的激活及其对细胞凋亡和病毒复制的影响仍存在争议。
研究 PPRV 诱导的 ER 应激以及 IRE1-XBP1 和 IRE1-JNK 通路的作用及其对细胞凋亡和病毒复制的影响。
通过 3-（4,5-二甲基-2-噻唑基）-2,5-二苯基-2H-四唑溴盐测定法、免疫荧光法和 Western blot 检测细胞活力和病毒复制。Western blot 和定量逆转录聚合酶链反应分别检测 ER 应激生物标志物 GRP78、IRE1 及其下游分子 PPRV-N 蛋白和凋亡相关蛋白的表达。分别使用 4-苯基丁酸（4-PBA）和 STF-083010 抑制 ER 应激和 IRE1 信号通路。
PPRV 感染细胞中 GRP78、IRE1α、p-IRE1α、XBP1s、JNK、p-JNK、caspase-3、caspase-9、Bax 和 PPRV-N 的表达明显上调，Bcl-2 的表达明显下调。由于 4-PBA 处理，GRP78、p-IRE1α、XBP1s、p-JNK、caspase-3、caspase-9、Bax 和 PPRV-N 的表达明显下调，Bcl-2 的表达明显上调。此外，在 PPRV 感染细胞中，p-IRE1α、p-JNK、Bax 和 PPRV-N 的表达降低，而在 STF-083010 存在的情况下，Bcl-2 的表达增加。
PPRV 感染诱导 ER 应激和 IRE1 激活，通过 IRE1-XBP1s 和 IRE1-JNK 通路诱导细胞凋亡并增强病毒复制。
