水疱性口炎病毒经AKT/Bax信号通路诱导神经细胞凋亡
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摘要
VSV神经嗜性是该病毒作为活病毒疫苗载体广泛应用的最大障碍,因此研究VSV诱导神经细胞的损伤对深入探讨VSV嗜神经性的分子机制尤为重要。本研究通过直接免疫荧光检测,可见VSV感染N2a细胞呈细胞核碎裂、染色质致密浓染等细胞凋亡的形态学变化;Annexin-V FITC/PI流式细胞术检测表明,接种VSV 8 h后N2a细胞凋亡率接近50%;Western blot结果显示在VSV感染过程中,磷酸化AKT表达量降低,Bax表达量升高。上述结果表明,VSV可通过AKT/Bax信号通路诱导神经细胞凋亡。
The neurotropism of VSV is the biggest obstace to its wide application as a live virus vaccine vector.Therefore,Exploring the mechanism of VSV-induced apoptosis of neuroblastoma cells was particularly important for the future study of neurotropism of VSV.The morphological changes including karyolysis and dense chromatin of apoptosis were observed by direct immunofluorescence assay.Flow cytometric detection with Annexin V-FITC/PI showed that the apoptosis rate of N2 a cells was nearly 50% after 8 h of VSV infection.Western blot results showed that AKT expression decreased and Bax expression increased during VSV infection.The results showed that VSV induced apoptosis of neuroblastoma cells through AKT/Bax signaling pathway,and the apoptosis induced by VSV was conducive to the replication and proliferation of VSV.
The neurotropism of VSV is the biggest obstace to its wide application as a live virus vaccine vector.Therefore,Exploring the mechanism of VSV-induced apoptosis of neuroblastoma cells was particularly important for the future study of neurotropism of VSV.The morphological changes including karyolysis and dense chromatin of apoptosis were observed by direct immunofluorescence assay.Flow cytometric detection with Annexin V-FITC/PI showed that the apoptosis rate of N2 a cells was nearly 50% after 8 h of VSV infection.Western blot results showed that AKT expression decreased and Bax expression increased during VSV infection.The results showed that VSV induced apoptosis of neuroblastoma cells through AKT/Bax signaling pathway,and the apoptosis induced by VSV was conducive to the replication and proliferation of VSV.
引文
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[2] VELAZQUEZSALINAS L,NAIK S,PAUSZEK S.Oncolytic recombinant vesicular stomatitis virus (VSV) is nonpathogenic and non-transmissible in pigs,a natural host of VSV[J].Hum Gene Ther,2017,28(2):85-90.
[3] 董雅洁,高维娟.bcl-2、bax、caspase-3在细胞凋亡中的作用及其关系[J].中国老年学杂志,2012,32(21):4828-4830.
[4] 信涛,赵妍,张磊,等.磁流体对肝癌细胞的凋亡诱导研究及联合紫杉醇对大鼠肝癌的抑制作用[J].中国肿瘤临床,2011,38(22):1363-1366.
[5] MICHAEL M,ARTURO L M,JENNIFER A.Oncolytic vesicular stomatitis virus as a viro-immunotherapy:defeating cancer with a “hammer” and “anvil”[J].Biomedicines,2017,5(4):8-11.
[6] SHINDE P V,XU H C,MANEY S K,et al.TNF-mediated survival of CD169+ cells promotes immune activation during vesicular stomatitis virus infection[J].J Virol,2018,92(3):JVI.01637-17.
[7] GADDY D F,LYLES D S.Vesicular stomatitis viruses expressing wild-type or mutant M proteins activate apoptosis through distinct pathways[J].J Virol,2005,79(7):4170.
[8] PAN W,SONG D,HE W,et al.EIF3i affects vesicular stomatitis virus growth by interacting with matrix protein[J].Vet Microbiol,2017,212:59-66.
[9] DUNN E F,CONNOR J H.Dominant inhibition of Akt/protein kinase B signaling by the matrix protein of a negative-strand RNA virus[J].Vet Microbiol,,2011,85(1):422-31.
[10] SOH T K,WHELAN S P J.Tracking the fate of genetically distinct vesicular stomatitis virus matrix proteins highlights the role for late domains in assembly[J].J Virol,2015:JVI.01371-15.
[11] 张强,崔媛旭,贾红豆,等.催乳素调控肝脏细胞脂代谢的信号机制[J].中国兽医学报,2018,38(3):570-575.
[12] DIEHL N,SCHAAL H.Make yourself at home:viral hijacking of the PI3K/Akt signaling pathway[J].Viruses,2013,5(12):3192.