抑制p38 MAPK信号通路对Bpiv3复制的影响
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摘要
由牛副流感病毒3型(Bovine parainfluenza virus type 3,Bpiv3)感染引起的牛副流感病已成为各国牛场最重要的传染病之一,每年都会给世界养牛业造成巨大的经济损失,但关于该病致病的分子机制研究较少。本研究通过观察Bpiv3感染对MDBK细胞中丝裂原活化蛋白激酶(MKK3)及其下游分子p38丝裂酶原活化的蛋白激酶(p38MAPK)的表达的影响,探讨相关的信号转导机制,对p38 MAPK通路在Bpiv3感染过程中的作用进行了初步研究。Bpiv3感染细胞后,采用Western Blot检测MKK3,p38 MAPK在蛋白水平的表达变化,并采用ELISA法检测细胞上清中IL-6,IL-8,IL-13和TNF-α的水平变化,采用SPSS 12软件进行统计学分析。结果表明,Bpiv3在感染后能够诱导MKK3的激活以及p38的磷酸化,激活了p38 MAPK信号通路。而且p38 MAPK信号通路参与了Bpiv3的复制过程。ELISA检测Bpiv3感染后以及使用抑制剂SB202190处理后的细胞上清中IL-6、IL-8、IL-13和TNF-α的水平发现,p38 MAPK信号通路参与了Bpiv3诱导的炎症反应。研究证实Bpiv3感染能够激活p38 MAPK通路,显著上调MKK3的表达并诱导p38发生磷酸化,进一步激活下游分子发挥生物学活性,促进Bpiv3的复制及诱导促炎细胞因子的产生。p38 MAPK信号通路的激活可能是Bpiv3感染诱发炎症反应的机制之一。
The bovine parainfluenza 3 virus( Bpiv3) can cause economic losses in the cattle industry. However,research on the molecular mechanism of the Bpiv3 infection are rarely reported. We measured expression of MKK3 and p38 mitogen-activated protein kinase( MAPK) after Bpiv3 infection. The signal-transduction mechanism of Bpiv3 infection was investigated, and the effect of the p38 MAPK pathway upon Bpiv3 infection was studied in a preliminary fashion. Expression of MKK3 and p38 MAPK was measured by Western Blot. The level of interleukin(IL)-6, IL-8, IL-13 and tumor necrosis factor(TNF)-α were tested by ELISA. Data were analyzed with SPSS 12. Results showed that Bpiv3 could induce activation of MKK3 and phosphorylation of p38 after infection. Bpiv3 infection could activate the p38 MAPK signaling pathway, and the latter, was involved in Bpiv3 replication. Levels of IL-6, IL-8, IL-13 and TNF-α, suggested that the p38 MAPK signaling pathway participated in the response to the inflammation caused by Bpiv3 infection. Activation of the p38 MAPK signaling pathway may be one of the mechanisms of inflammation induced by Bpiv3 infection.
The bovine parainfluenza 3 virus( Bpiv3) can cause economic losses in the cattle industry. However,research on the molecular mechanism of the Bpiv3 infection are rarely reported. We measured expression of MKK3 and p38 mitogen-activated protein kinase( MAPK) after Bpiv3 infection. The signal-transduction mechanism of Bpiv3 infection was investigated, and the effect of the p38 MAPK pathway upon Bpiv3 infection was studied in a preliminary fashion. Expression of MKK3 and p38 MAPK was measured by Western Blot. The level of interleukin(IL)-6, IL-8, IL-13 and tumor necrosis factor(TNF)-α were tested by ELISA. Data were analyzed with SPSS 12. Results showed that Bpiv3 could induce activation of MKK3 and phosphorylation of p38 after infection. Bpiv3 infection could activate the p38 MAPK signaling pathway, and the latter, was involved in Bpiv3 replication. Levels of IL-6, IL-8, IL-13 and TNF-α, suggested that the p38 MAPK signaling pathway participated in the response to the inflammation caused by Bpiv3 infection. Activation of the p38 MAPK signaling pathway may be one of the mechanisms of inflammation induced by Bpiv3 infection.
引文
[1] Grissett G P,White B J, Larson R L.Structured literature review of responses of cattle to viral and bacterial pathogens causing bovine respiratory disease complex[J]. J Vet Intern Med, 2015, 29(3):770-780.
[2] Solis-Calderon J J,Segura-Correa J C,Aguilar Romero F,Segura-Correa V M.Detection of antibodies and risk factors for infection with bovine respiratory syncytial vi-rus and parainfluenza virus-3 in beef cattle of Yucatan,Mexico[J]. Prev Vet Med, 2007, 82(1-2):102-110.
[3] Horwood P F, Mahony T J. Multiplex real-time RTPCR detection of three viruses associated with the bovine respiratory disease complex[J]. J Virol Methods,2011,171(2):360-363.
[4] Kirchhoff J,Uhlenbruck S,Goris K,Keil G M,Herrler G. Three viruses of the bovine respiratory disease complex apply different strategies to initiate infection[J]. Vet Res, 2014, 45(1):20-32.
[5]张频捷,朱立新,耿小平.p38MAPK信号传导通路及其抑制剂的研究现状[J].安徽医药,2010, 14(5):596-598.
[6]孙晓林,杜方原,刘淑英.外源性绵羊肺腺瘤病毒囊膜蛋白激活MAPK信号通路的分析[J].畜牧兽医学报,2016, 47(8):1658-1666.
[7]丁利,黄勇,李兆才,戴美玲,于高水,杜谦,童德文.猪传染性胃肠炎病毒感染诱发ROS的积累、p38和p53信号的活化[C].中国畜牧兽医学会兽医病理学分会暨中国病理生理学会动物病理生理专业委员会学术研讨会,2012.
[8]蒲长宇.乙型肝炎病毒X蛋白对肝癌细胞MKK3,p38MAPK表达的影响[J].现代肿瘤医学,2011,19(4):640-644.
[9] Choi M S, Heo J, Yi C M, Ban J, Lee N J, Lee N R,Kim S W,Kim N J,Inn KSet al.A novel p38 mitogen activated protein kinase(MAPK)specific inhibitor suppresses respiratory syncytial virus and influenza A virus replication by inhibiting virus-induced p38 MAPK activation[J]. Biochem Biophys Res Commun, 2016,477(3):311-316.
[10] Lisnock J,Tebben A,Frantz B,O'Neill EA,Croft G,O'Keefe SJ,Li B,Hacker C,de Laszlo S, Smith A,Libby B, Liverton N, Hermes J, LoGrasso P.Molecular basis for p38 protein kinase inhibitor specificity[J]. Biochem,1998, 37(47):16573-16581.
[11] Ellis J A. Bovine parainfluenza-3 virus[J]. Vet Clin North Am Food Anim Pract,2010,26(3):575-593.
[12] Weifeng Shi, Xueling Houa,Xiang Li,Hongjun Peng,Mei Shia, Qingbo Jianga, Xiping Liu, Yun Ji, Yuhua Yao, Caizhen He, Xiangdong Lei. Differential gene expressions of the MAPK signaling pathway in enterovirus71-infected rhabdomyosarcoma cells[J]. Braz J Infect Dis, 2013,17(4):410-417.
[13] Si X,Luo H,Morgan A,Zhang J,Wong J,Yuan J,Esfandiarei M, Gao G, Cheung C, McManus M.Stressactivated protein kinases are involved in Coxsackievirus B3 viral progeny release[J]. J Virol,2005,79(22):13875-13881.
[14] Suarez C J, Parker N J, Finn P W.Innate immune mechanism in allergic asthma[J]. Curr Allergy Asthma Rep,2008, 8(5):451-459.
[15] Lee C,Kim Y,Ji H J.JNK and p38 mitogen-activated protein kinase pathways contribute to porcine epidemic diarrhea virus infection[J]. Virus Res, 2016, 222:1-12.
[16] Donovan C,Bourke J E,Vlahos R.Targeting the IL-33/IL-13 axis for respiratory viral infections[J]. Trens Pharmacol Sci, 2016, 37(4):252-261.
[17] Peifer C,Wagner G,Laufer S.New approaches to the treatment of inflammatory disorders small molecule inhibitors of p38 MAP kinase[J]. Curr Top Med Chem,2006, 6(2):113-149.
[2] Solis-Calderon J J,Segura-Correa J C,Aguilar Romero F,Segura-Correa V M.Detection of antibodies and risk factors for infection with bovine respiratory syncytial vi-rus and parainfluenza virus-3 in beef cattle of Yucatan,Mexico[J]. Prev Vet Med, 2007, 82(1-2):102-110.
[3] Horwood P F, Mahony T J. Multiplex real-time RTPCR detection of three viruses associated with the bovine respiratory disease complex[J]. J Virol Methods,2011,171(2):360-363.
[4] Kirchhoff J,Uhlenbruck S,Goris K,Keil G M,Herrler G. Three viruses of the bovine respiratory disease complex apply different strategies to initiate infection[J]. Vet Res, 2014, 45(1):20-32.
[5]张频捷,朱立新,耿小平.p38MAPK信号传导通路及其抑制剂的研究现状[J].安徽医药,2010, 14(5):596-598.
[6]孙晓林,杜方原,刘淑英.外源性绵羊肺腺瘤病毒囊膜蛋白激活MAPK信号通路的分析[J].畜牧兽医学报,2016, 47(8):1658-1666.
[7]丁利,黄勇,李兆才,戴美玲,于高水,杜谦,童德文.猪传染性胃肠炎病毒感染诱发ROS的积累、p38和p53信号的活化[C].中国畜牧兽医学会兽医病理学分会暨中国病理生理学会动物病理生理专业委员会学术研讨会,2012.
[8]蒲长宇.乙型肝炎病毒X蛋白对肝癌细胞MKK3,p38MAPK表达的影响[J].现代肿瘤医学,2011,19(4):640-644.
[9] Choi M S, Heo J, Yi C M, Ban J, Lee N J, Lee N R,Kim S W,Kim N J,Inn KSet al.A novel p38 mitogen activated protein kinase(MAPK)specific inhibitor suppresses respiratory syncytial virus and influenza A virus replication by inhibiting virus-induced p38 MAPK activation[J]. Biochem Biophys Res Commun, 2016,477(3):311-316.
[10] Lisnock J,Tebben A,Frantz B,O'Neill EA,Croft G,O'Keefe SJ,Li B,Hacker C,de Laszlo S, Smith A,Libby B, Liverton N, Hermes J, LoGrasso P.Molecular basis for p38 protein kinase inhibitor specificity[J]. Biochem,1998, 37(47):16573-16581.
[11] Ellis J A. Bovine parainfluenza-3 virus[J]. Vet Clin North Am Food Anim Pract,2010,26(3):575-593.
[12] Weifeng Shi, Xueling Houa,Xiang Li,Hongjun Peng,Mei Shia, Qingbo Jianga, Xiping Liu, Yun Ji, Yuhua Yao, Caizhen He, Xiangdong Lei. Differential gene expressions of the MAPK signaling pathway in enterovirus71-infected rhabdomyosarcoma cells[J]. Braz J Infect Dis, 2013,17(4):410-417.
[13] Si X,Luo H,Morgan A,Zhang J,Wong J,Yuan J,Esfandiarei M, Gao G, Cheung C, McManus M.Stressactivated protein kinases are involved in Coxsackievirus B3 viral progeny release[J]. J Virol,2005,79(22):13875-13881.
[14] Suarez C J, Parker N J, Finn P W.Innate immune mechanism in allergic asthma[J]. Curr Allergy Asthma Rep,2008, 8(5):451-459.
[15] Lee C,Kim Y,Ji H J.JNK and p38 mitogen-activated protein kinase pathways contribute to porcine epidemic diarrhea virus infection[J]. Virus Res, 2016, 222:1-12.
[16] Donovan C,Bourke J E,Vlahos R.Targeting the IL-33/IL-13 axis for respiratory viral infections[J]. Trens Pharmacol Sci, 2016, 37(4):252-261.
[17] Peifer C,Wagner G,Laufer S.New approaches to the treatment of inflammatory disorders small molecule inhibitors of p38 MAP kinase[J]. Curr Top Med Chem,2006, 6(2):113-149.