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Abstract：
BACKGROUND: 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.
OBJECTIVE: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: PPRV infection induces ER stress and IRE1 activation, resulting in apoptosis and enhancement of virus replication through IRE1-XBP1s and IRE1-JNK pathways.
OBJECTIVE: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: PPRV infection induces ER stress and IRE1 activation, resulting in apoptosis and enhancement of virus replication through IRE1-XBP1s and IRE1-JNK pathways.
摘要：
背景：小反刍动物（PPR）是绵羊和山羊的传染性和致命性疾病。PPR病毒(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。
目的：研究PPRV诱导的ER应激和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。
方法：通过3-（4,5-二甲基-2-噻唑基）-2,5-二苯基-2H-四唑溴化物测定法评估细胞活力和病毒复制，免疫荧光测定，和Westernblot。ER应激生物标志物GRP78、IRE1及其下游分子的表达,PPRV-N蛋白,通过Westernblot和定量逆转录聚合酶链反应检测凋亡相关蛋白，分别。4-苯丁酸(4-PBA)和STF-083010分别用于抑制ER应激和IRE1信号通路。
结果：GRP78、IRE1α、p-IRE1α,XBP1,JNK,p-JNK,caspase-3、caspase-9、Bax和PPRV-N在PPRV感染细胞中显著上调,Bcl-2的表达显著下调。由于4-PBA治疗，GRP78，p-IRE1α的表达，XBP1,p-JNK,caspase-3,caspase-9,Bax,PPRV-N显著下调，Bcl-2的表达显著上调。此外，在PPRV感染的细胞中，p-IRE1α的表达，p-JNK,Bax,PPRV-N显著下降，在STF-083010存在下Bcl-2的表达增加。
结论：PPRV感染诱导ER应激和IRE1激活，通过IRE1-XBP1s和IRE1-JNK途径导致细胞凋亡和病毒复制增强。
目的：研究PPRV诱导的ER应激和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。
方法：通过3-（4,5-二甲基-2-噻唑基）-2,5-二苯基-2H-四唑溴化物测定法评估细胞活力和病毒复制，免疫荧光测定，和Westernblot。ER应激生物标志物GRP78、IRE1及其下游分子的表达,PPRV-N蛋白,通过Westernblot和定量逆转录聚合酶链反应检测凋亡相关蛋白，分别。4-苯丁酸(4-PBA)和STF-083010分别用于抑制ER应激和IRE1信号通路。
结果：GRP78、IRE1α、p-IRE1α,XBP1,JNK,p-JNK,caspase-3、caspase-9、Bax和PPRV-N在PPRV感染细胞中显著上调,Bcl-2的表达显著下调。由于4-PBA治疗，GRP78，p-IRE1α的表达，XBP1,p-JNK,caspase-3,caspase-9,Bax,PPRV-N显著下调，Bcl-2的表达显著上调。此外，在PPRV感染的细胞中，p-IRE1α的表达，p-JNK,Bax,PPRV-N显著下降，在STF-083010存在下Bcl-2的表达增加。
结论：PPRV感染诱导ER应激和IRE1激活，通过IRE1-XBP1s和IRE1-JNK途径导致细胞凋亡和病毒复制增强。
