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通过光镜、电镜、DNA Ladder法、流式细胞术、荧光染色对鸭呼肠孤病毒(DRV)诱导鸭胚原代成纤维细胞(DEF)凋亡情况进行检测。结果显示,光镜可见细胞形态学上出现细胞皱缩,染色质浓染边移;电镜观察到细胞胞浆浓缩,细胞核染色质凝聚、部分形成凋亡小体;荧光染色结果显示,在感染后24h有激发绿色荧光的凋亡细胞出现,随着时间的推移,激发红色荧光的死亡细胞数量增多;DNA Ladder检测到感染后24~144h的DNA样品呈梯形条带;流式细胞术于感染后24h检测到凋亡细胞,其数量在72~96h达到高峰,144h开始下降。研究结果表明,DRV在DEF增殖的过程中具有诱导宿主细胞凋亡的作用。
Abstract：Cell apoptosis induced by duck reovirus (DRV)in duck embryo fibroblasts (DEF) was ascertained by light microscope and electron microscopy, DNA Ladder, flow cytometry and fluorescent microscopy. Typical morphological apoptotic features including cell shrinkage and condensation, margination of nuclear chromatin were observed under light microscope and the formation of apoptotic bodies by electron microscopy. DNA ladder was shown by DNA fragment analysis at 24-144h post infection. Flow cytometry showed that the cell apoptosis appeared at 24h and reached it's crest-time at 72-96h, decreased at 144h. Fluorescent microscopy showed that the apoptotic cells which showed green fluores-cence appeared at 24h, the number of dead cells which showed red fluores-cence increased with the time went by. The results above confirmed that the apoptosis of DEF was successfully induced by DRV.
[1]程安春,汪铭书,陈孝跃,等.一种新发现的鸭病毒性肿头出血症的研究[J].中国兽医科技,2003,33(10):15-21.
[2]Peter M,Heufelder H,Hengartner M.Advances in ap-optosis research[J].Proc Natl Acad Sci USA,1997,94(1):12736-12737.
[3]Obrien V.Viruses and apoptosis[J].J Gen Virol,1998,79(3):33-1845.
[4]张娜,程安春,汪铭书,等.鸭肿头出血症病毒在鸭胚原代成纤维细胞中增值特性的研究[J].中国兽医科学,2007,37(9):746-750.
[5]殷震,刘景华.动物病毒学[M].2版.北京:科学出版社,1997:544-548.
[6]Taeg S K,David A,Benfield B,et al.Porcine reproduc-tive and respiratory syndrome virus-induced cell death ex-hibits features consistent with a nontypical formof apop-tosis[J].Virus Res,2002,85(5):133-140.
[7]Micoud F,Mandrand B,Malcus V C.Comparison of several techniques for the detection of apoptotic astro-cytesin vitro[J].Cell Prolif,2001,34(2):99-133.
[8]Wang H,Wang Z,Chen J,et al.Apoptosis induced by no via phosphorylation of p38MAPKthat sti mulates NF-κB,p53and caspase-3activationin rabbit articular chon-drocytes[J].Cell Biol Int,2007,31(9):1027-1035.
[9]Schobesberger M,Summerfield A,Doherr M G,et al.Canine distemper virus-induced depletion of uninfectedlymphocytes is associated with apoptosis[J].Vet I mmu-nol I mmunopathol,2005,104(1-2):33-44.
[10]Labrada L,Bodelon G,Vinuela J,et al.Avian reovi-ruses cause apoptosis in cultured cells:viral uncoating,but not viral gene expression,is required for apoptosis induction[J].J Virol,2002,76(16):7932-7941.
[11]Salsman J,Top D,Boutilier J,et al.Extensive syncy-tiumformation mediated by the reovirus FASTproteins triggers apoptosis-induced membrane instability[J].J Virol,2005,79(13):8090-8100.
[12]Coffey C M,Sheh A,Ki mI S,et al.Reovirus outer capsid proteinμ1induces apoptosis and associates with lipid droplets,endoplasmic reticulum,and mitochon-dria[J].J Virol,2006,80(17):8422-8438.
[13]Clarke P,Tyler K L.Reovirus-induced apoptosis:a mini-review[J].Apoptosis,2003,8(2):141-150.
[14]DeBiasi R L,Robinson B A,Sherry B,et a1.Caspase inhibition protects against reovirus-induced myocardial injuryin vitroandin vivo[J].J Virol,2004,78(20):11040-11050.
[15]DeBiasi R L,Edelstein C L,Sherry B,et a1.Calpaln inhibition protects against virus-induced apoptotic myo-cardial injury[J].J Virol,2001,75(11):351-361.
[16]Fahey J E,Crawley J F.Studies on chronic respiratory disease of chickens II.Isolation of a virus[J].Can J Comp Med,1954,18:13-21.
[17]Hieronymus D R K,Villegas P,Kleven S H.Identifi-cation and serological differentiation of several reovirus strains isolated from chickens with suspected malab-sorption syndrome[J].Avian Dis,1983,27(20):246-254.
[18]Kibenege F S B,Wilcox G E.Tenosynovitis in chick-ens[J].Vet Bull,1983,53(7):431-443.
[19]Roessler D E,Rosenberger J K.In vitroandin vivo characterization of avian reoviruses III.Host factors affecting virulence and persistence[J].Avian Dis,1989,33,(6):555-565.
[20]Shih WL,Hsu H W,Liao M H,et al.Avian reovirusσC proteininduces apoptosis in cultured cells[J].Vi-rology,2004,321(1):65-74.
[21]Wyllie A H,Kerr J F,Currie A R.Cell death:the significance of apoptosis[J].Int Rev Cytol,1980,8(5):251-306.
[22]Rosl F A.Si mple and rapid method for detection of ap-optosis in human cells[J].Nucl Acids Res,1992,20(19):5243.
[23]Barbouti A,Doulias P T,Nousis L,et al.DNAdam-age and apoptosis in hydrogen peroxide exposed Jurkat cells:bolus addition versus continuous generation of H2O2[J].Free Radic Biol Med,2002,33(11):691-702.
[24]Yasuhara S,Kanakubo E,Perez M E,et al.Burn in-jury induces skeletal muscle apoptosis and the activa-tion of caspase pathways in rats[J].J Burn Care Re-hab,1999,20(9):462-470.
[25]Fadok V A,Voelker D R,Campbell P A,et al.Expo-sure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages[J].J I mmunol,1992,148(7):2207-2216.
[26]Andree H A M,Reutelingsperger C P M,Haupt man R,et al.Binding of vascular anticoagulant a(VACa)to planar phospholipid bilayers[J].J Biol Chem,1990,265(5):4923.
[27]Thomson B J.Viruses and apoptosis[J].Int J Exp Pathol,2001,82(2):65-76.
基本信息:
DOI：10.13242/j.cnki.bingduxuebao.001908
中图分类号:S858.32
引用信息:
[1]张娜,程安春,汪铭书,等.鸭呼肠孤病毒强毒株致鸭胚原代成纤维细胞凋亡的研究[J].病毒学报,2008(03):213-219.DOI:10.13242/j.cnki.bingduxuebao.001908 .
基金信息:
国家“十一”科技支撑计划(2007Z06-017); 教育部“新世纪优秀人才支持计划”项目(NCET-04-0906/NCET-06-0818); 四川省杰出青年基金(07ZQ026-132); 四川省重大基础研究项目(04JY029-006-1); 四川省重点建设学科项目(SZD0418)
2008-05-15
2008-05-15