Hyper-IL-6重组腺病毒介导急性肝衰竭治疗的实验研究
|
摘要
目的:构建Hyper-IL-6重组腺病毒质粒及作为治疗对照的IL-6重组腺病毒、空腺病毒质粒,进行病毒的包装扩增,并探讨重组腺病毒介导的Hyper-IL-6对大鼠急性肝衰竭的治疗作用及机制。
方法:将腺病毒穿梭质粒pAdTrack-Hyper-IL-6以Pme-I酶切线性化后与腺病毒骨架质粒AdEasy-1在BJ5183菌内进行同源重组;经酶切及PCR鉴定构建成功后,以Pac-I酶切Hyper-IL-6重组腺病毒(AdHIL-6)质粒,将线性化的AdHIL-6质粒经脂质体转染293细胞进行腺病毒的包装扩增,以氯化铯密度梯度离心法纯化病毒,同法制备AdIL-6、Ad0重组腺病毒。以D-氨基半乳糖(D-Gal)诱导急性肝衰竭模型,诱导后6h,80只SD大鼠随机分为4组:未感染组、Ad0感染组、AdIL-6感染组、AdHIL-6感染组;每组大鼠又分为A、B两组:A组观察3d,采血进行肝功能指标检测,取肝组织H-E染色观察病理学变化,原位末端标记法(TUNEL法)检测肝细胞凋亡,免疫组织化学法检测增殖细胞核抗原(PCNA)、Caspase-3在肝组织中的表达;B组留观14d存活率。
结果:重组腺病毒质粒经Pac-I酶切后可见3kb和一条约30kb的条带,表明同源重组成功。将线性化的重组腺病毒质粒转染293细胞后于荧光显微镜下可观察到明亮的绿色荧光,收集的病毒液经PCR扩增出IL-6、Hyper-IL-6和AdEasy-1特异性片段。AdHIL-6感染组血清丙氨酸氨基转移酶(ALT)、总胆红素(TB)含量及凋亡指数明显低于未感染组、Ad0感染组及AdIL-6感染组(P<0.01),而肝组织PCNA表达明显高于另3组(P<0.01);未感染组及Ad0感染组肝组织坏死严重,Caspase-3极强阳性表达;AdIL-6感染组肝组织病变稍轻,Caspase-3强阳性表达;AdHIL-6感染组肝组织病变明显减轻,Caspase-3表达阳性。观察14d后,未感染组及Ad0感染组大鼠全部死亡,AdIL-6感染组1只存活,AdHIL-6感染组存活3只。
结论:成功地完成了3种重组腺病毒(AdHIL-6、AdIL-6、Ad0)的制备;Hyper-IL-6对急性肝衰竭大鼠有明显的促肝细胞增殖及抗肝细胞凋亡效应,能有效改善肝功能及肝组织学状况,提高肝衰竭大鼠的存活率。
Objective:To construct the adenoviral vector that expresses fusion gene encoding Hyper-IL-6,and the controls that express IL-6 or adenovirus not containing objective gene.To package and propogate the three recombinant adenoviruses respectively ,then to explore the treatment effects and mechanisms of Hyper-IL-6 mediated by recombinant adenovirus on rat with acute liver failure(ALF).
Methods:The adenoviral shuttle plasmid containing Hyper-IL-6 was digested with Pme-I.The linearized shuttle plasmids were co-transformed with adenoviral backbone vector to E.coli BJ5183 cells. The candidate clones were further tested by restriction nuclease digestion with PacⅠand PCR. Before transfection, the recombinant adenoviral plasmids were digested with PacI. Then the digested recombinant adenoviral plasmids were transfected to 293 cells with liposome for package and amplification of adenoviruses,then the viruses were purified through CsCl density-gradient centrifugalization .The control viruses were generated with the same method. The ALF rat model was induced by D-Gal ,and eighty SD rats were randomly divided into 4 groups(each 20) after 6h injected D-Gal:normal saline was injected for non-infection group ;Ad0,AdIL-6 and AdHIL-6 was respectively injected for group Ad0,group AdIL-6,group AdHIL-6.Sequently each group was again divided into group A(n=10) and group B(n=10).Group A was observed for 3 days,then the blood samples were collected to determine the index of hepatic function(ALT,TB),the liver tissues were taken for H-E staining,immunohistochemistry of PCNA and Caspase-3,and to detect the hepatocelluar apoptosis in situ by TUNEL assay.Group B was kept observing for 14 days in order to conclude the survival rate of rats.
Results:The object staps about 3kb and 30kb emerged after the recombinant adenoviral plasmids were digested with Pac-I,which demonstrated successfully homologous recombination.After transfection, the brighter green fluorescence was observed in 293 cells.The viruse liquid can be amplified object genes:IL-6,Hyper-IL-6,AdEasy-1.The contents of serum ALT and TBil,and apoptotic index in group AdHIL-6 were markedly decreased than that of the other groups(P<0.01),inversely the significant expression of PCNA(P<0.01) .The state of liver necrosis was terrible,and extremely positive expression of Caspase-3 was in non-infection group and group Ad0.The extent to liver injury was slightly reduced than that of the former 2 groups,and highly positive expression of Caspase-3 in group AdIL-6.The liver pathological change was the most mild,and positive expression of Caspase-3 in group AdHIL-6.At the end of 14 days after injecting D-Gal,all rats died in non-infection group and group Ad0,there was only one alive in group AdIL-6 ,however 3 rats stayed lively in group AdHIL-6.
Conclusion:The generating of the three recombinant adenoviruses was successful. It was significant that the potency of Hyper-IL-6 enhancing hepatocelluar proliferation and defending hepatocyte apoptosis on the rats with ALF. Hyper-IL-6 can effectively ameliorate liver function and histology,and improve the survival of rats with ALF.
方法:将腺病毒穿梭质粒pAdTrack-Hyper-IL-6以Pme-I酶切线性化后与腺病毒骨架质粒AdEasy-1在BJ5183菌内进行同源重组;经酶切及PCR鉴定构建成功后,以Pac-I酶切Hyper-IL-6重组腺病毒(AdHIL-6)质粒,将线性化的AdHIL-6质粒经脂质体转染293细胞进行腺病毒的包装扩增,以氯化铯密度梯度离心法纯化病毒,同法制备AdIL-6、Ad0重组腺病毒。以D-氨基半乳糖(D-Gal)诱导急性肝衰竭模型,诱导后6h,80只SD大鼠随机分为4组:未感染组、Ad0感染组、AdIL-6感染组、AdHIL-6感染组;每组大鼠又分为A、B两组:A组观察3d,采血进行肝功能指标检测,取肝组织H-E染色观察病理学变化,原位末端标记法(TUNEL法)检测肝细胞凋亡,免疫组织化学法检测增殖细胞核抗原(PCNA)、Caspase-3在肝组织中的表达;B组留观14d存活率。
结果:重组腺病毒质粒经Pac-I酶切后可见3kb和一条约30kb的条带,表明同源重组成功。将线性化的重组腺病毒质粒转染293细胞后于荧光显微镜下可观察到明亮的绿色荧光,收集的病毒液经PCR扩增出IL-6、Hyper-IL-6和AdEasy-1特异性片段。AdHIL-6感染组血清丙氨酸氨基转移酶(ALT)、总胆红素(TB)含量及凋亡指数明显低于未感染组、Ad0感染组及AdIL-6感染组(P<0.01),而肝组织PCNA表达明显高于另3组(P<0.01);未感染组及Ad0感染组肝组织坏死严重,Caspase-3极强阳性表达;AdIL-6感染组肝组织病变稍轻,Caspase-3强阳性表达;AdHIL-6感染组肝组织病变明显减轻,Caspase-3表达阳性。观察14d后,未感染组及Ad0感染组大鼠全部死亡,AdIL-6感染组1只存活,AdHIL-6感染组存活3只。
结论:成功地完成了3种重组腺病毒(AdHIL-6、AdIL-6、Ad0)的制备;Hyper-IL-6对急性肝衰竭大鼠有明显的促肝细胞增殖及抗肝细胞凋亡效应,能有效改善肝功能及肝组织学状况,提高肝衰竭大鼠的存活率。
Objective:To construct the adenoviral vector that expresses fusion gene encoding Hyper-IL-6,and the controls that express IL-6 or adenovirus not containing objective gene.To package and propogate the three recombinant adenoviruses respectively ,then to explore the treatment effects and mechanisms of Hyper-IL-6 mediated by recombinant adenovirus on rat with acute liver failure(ALF).
Methods:The adenoviral shuttle plasmid containing Hyper-IL-6 was digested with Pme-I.The linearized shuttle plasmids were co-transformed with adenoviral backbone vector to E.coli BJ5183 cells. The candidate clones were further tested by restriction nuclease digestion with PacⅠand PCR. Before transfection, the recombinant adenoviral plasmids were digested with PacI. Then the digested recombinant adenoviral plasmids were transfected to 293 cells with liposome for package and amplification of adenoviruses,then the viruses were purified through CsCl density-gradient centrifugalization .The control viruses were generated with the same method. The ALF rat model was induced by D-Gal ,and eighty SD rats were randomly divided into 4 groups(each 20) after 6h injected D-Gal:normal saline was injected for non-infection group ;Ad0,AdIL-6 and AdHIL-6 was respectively injected for group Ad0,group AdIL-6,group AdHIL-6.Sequently each group was again divided into group A(n=10) and group B(n=10).Group A was observed for 3 days,then the blood samples were collected to determine the index of hepatic function(ALT,TB),the liver tissues were taken for H-E staining,immunohistochemistry of PCNA and Caspase-3,and to detect the hepatocelluar apoptosis in situ by TUNEL assay.Group B was kept observing for 14 days in order to conclude the survival rate of rats.
Results:The object staps about 3kb and 30kb emerged after the recombinant adenoviral plasmids were digested with Pac-I,which demonstrated successfully homologous recombination.After transfection, the brighter green fluorescence was observed in 293 cells.The viruse liquid can be amplified object genes:IL-6,Hyper-IL-6,AdEasy-1.The contents of serum ALT and TBil,and apoptotic index in group AdHIL-6 were markedly decreased than that of the other groups(P<0.01),inversely the significant expression of PCNA(P<0.01) .The state of liver necrosis was terrible,and extremely positive expression of Caspase-3 was in non-infection group and group Ad0.The extent to liver injury was slightly reduced than that of the former 2 groups,and highly positive expression of Caspase-3 in group AdIL-6.The liver pathological change was the most mild,and positive expression of Caspase-3 in group AdHIL-6.At the end of 14 days after injecting D-Gal,all rats died in non-infection group and group Ad0,there was only one alive in group AdIL-6 ,however 3 rats stayed lively in group AdHIL-6.
Conclusion:The generating of the three recombinant adenoviruses was successful. It was significant that the potency of Hyper-IL-6 enhancing hepatocelluar proliferation and defending hepatocyte apoptosis on the rats with ALF. Hyper-IL-6 can effectively ameliorate liver function and histology,and improve the survival of rats with ALF.
引文
[1] Polson J, Lee WM, American Ascociation for the study of Liver Disease. AASLD position paper: the management of acute liver failure [J].Hepatology, 2005, 41(5): 1179- 1197.
[2] Fausto NJ.Liver regeneration[J].Hepatol,2000,32:19-31.
[3] Fischer M, Goldschmitt J, Peschel C,et al. A bioactive designer cytokine for human hemotopoietic progenitor cell expansion[J].Nature Biotechnol,1997,15:142-145.
[4] Galun E, Axelrod JH. The role of cytokines in liver failure and regeneration: potential new therapies. Biochim Biophys Acta, 2002,1592(3):354-358.
[5]郭国宁,周跃,张正丰.成骨细胞特异性转录因子 Cbfa1 重组腺病毒的构建、纯 化及表达[J].免疫学杂志.2005,21(2):150-154.
[6] D.L.斯配克特,R.D.戈德曼,L.A.莱因万德.细胞试验指南[M].黄培堂等译. 北京: 科学出版社,2001:830-831.
[7] 董世武,应大君,刘光久,等.小鼠 Osf2/Cbfal 基因的重组腺病毒构建及鉴定 [J ]. 第三军医大学学报. 2003, 25 (21) : 1947—1954.
[8] K.K.杰恩, 基因治疗学[M].任斌等译.西安:世界图书出版西安公司,2000: 603
[9] Kinoshita K,limuro Y,Fujimoto J,et a1.Targeted and regulable expression of transgenes in hepatic stellate cells and myofibroblasts in culture and in vivo using an adenoviral Cre /loxP system to antagonise hepatic fibrosis[J].Gut,2007,56(3):396 -404.
[10] Liu L,Pedersen E F. Partial abstracts of the 3rd annual meeting of the China RNA society & 1st international RNA workshop in China:SESSION1:mRNA processing biochemical identification of nuclear protein p30 specifically interacting with upstrea- m repressi[J]. 生物化学与生物物理学报(英文版),2004,36(2):147
[11] Kalyuzhniy O,Di Paolo NC,Si lvestry M,et al.Adenovirus serotype 5 hexon is critic- al for virus infection of hepatocytes in vivo[J]. Proc Natl Acad Sci U S A.2008,105 (14):5483-5488.
[12] Robert JJ,Gauffeny I,Maccario J,et al.Degenerated pIX-IVa2adenoviral vectorsequences lowers reacquisition of the E1genes duringvirus amplification in293cells[J].Gene Ther,2001,8(22):1713-1720.
[13]Crouzet J,Naudin L,Orsini c,et a1.Recombinational construction in escherichia coli of infectious adenoviral genomes[J].ProcAcad Sci USA.1997,94(4):1414—1419.
[14]Davis AR,Wivel NA,Palladino JL,et a1.Construction of adenoviral vectors [J].Mol Biotechnol,2001,18(1):63—70.
[15] He TC, Zhou S, da Costa LT, et al. A simplified system for generating recombinant adenoviruses[J]. Proc Natl Acad Sci U S A. 1998 ,95(5):2509-14
[16]CONG Tie-chuan,LU Zhe-ming,Li Yong,et al.High efficient generation of replication-defective adenoviruses containing thymidine kinase by homogeneous recombination in bacteria[J].Chin Med . 2007,120(18):1622-1625.
[17]吕慧,赵蔚明,于修平,等. E 型沙眼衣原体 MOMP 基因重组腺病毒的构建及免疫原性研究[J].山东大学学报(医学版),2006,44(12):1189-1193.
[18]李玉峰,姜平,蒋文明,等.猪繁殖与呼吸综合征病毒 GP5 蛋白重组腺病毒的构建与免疫原性测定[J].中国病毒学,2006,21(4):364-367.
[19]Kalyanasundaram S,Feinstein S,Nicholson JP, et al. Coacervate microspheres as carriers of recombinant adenoviruses[J].Cancer Gene Ther, 1999, 6(2): 107-112.
[20]中华医学会传染病与寄生虫病学分会、肝病学分会联合修订.病毒性肝炎防治方案[J].中华肝脏病杂志,2000,8(6):324-329.
[21] Zimmers TA,Pierce RH,Mckillop IH,et al. Resolving the role of IL-6 in liver regeneration [J].Hepatol,2003,38:1590-1591.
[22] Boulanger MJ,Chow D,Brevnova EE,et al.Hexameric structure and assembly of the interleukin-6/IL-6a-receptor/gp130 complex[J].Science,2003,300:2101-2104.
[23] Oberg HH,Wesch D,Grussel S,et al. Differential expression of CD126 and CD130 mediates different STAT-3 phosphorylation in CD4+CD25- and CD25high regulatory T cells[J].Int Immunol,2006,18(4):555-563.
[24] Igaz P,Horvath A,Horvath B,et al. Soluble interleukin-6 receptor (sIL-6R) makes IL-6R negative T cell line respond to IL-6;it inhibits TNF production[J].ImmunolLetter,2000,71:143-148.
[25]Jones SA,Richards PJ,Scheller J,et al.IL-6 transsignaling:the in vivo consequences[J].Interferon Cytokine Res 2005(25):241-253.
[26] Zvibel I, Brill S, Kevital R, et al. Chimeric molecule IL-6/soluble receptor is a potent mitogen for fetal hepatocytes[J] .Cell Physiol,2004,200:245-252.
[27] Peters M, Solem F, Schirmacher P, et al. IL-6 and soluble IL-6R induce stem cell factor (SCF) and Flt-3 ligand expression in vivo and in vitro[J]. Exp Hematol, 2001, 29:146–155.
[28] Peters M, Blinn G, Jostock T,et al. Combined interleukin 6 and soluble interleukin 6 receptor accelerates murine liver regeneration[J].Gastroenterol,2000,119(6):1163 –1171.
[29] Male Peters,Guido Blinn,Fian Solem,et al.In vivo and in vitro activities of the gp130-stimulating designer cytokine Hyper-IL-6[J]. Immunol,1998,161:3575-3581.
[30] Yeoh GC,Ernst M,Rose-J0hn S,Akhurst B,et al.Opposing roles of gp130-mediated STAT-3and ERK-1/2signaling in liver progenitor cell migration and proliferation[J].Hepatoology,2007,45(2):486-494.
[31] Richards PJ,Nowell MA,Horiuchi S,et al.Functional characterization of a soluble gp130 isform and its therapeutic capacity in an experimental model of inflammatory arthritis[J].Arthritis Rheum,2006,54(5):1662-1672.
[32] Hecht N,Pappo O,Shouval D,et al.Hyper-IL-6 gene therapy reverses fulminant hepatic failure[J].Mol Therap ,2001,3:683-687.
[33]Gotze KS, Keller U, Rose-John S, et al. Gp130-stimulating designer cytokine hyper-interleukin-6 synergizes with murine stroma for long-term survival of primitive human hematopietic progenitor cells[J]. Exp Hematol, 2001,29(7): 822-32
[34] Wang XP, Schunck M, Kallen KJ, et al. The interleukin-6 cytokine system regulates epidermal permeability barrier homeostasis[J]. J Invest Dermatol.2004 Jul;123(1):124-31
[35] Sun Y, Marz P, Otten U,et al. The effect of gp130-stimulation on glutamate-induced excitotoxicity in primary hippocampal neurons[J]. Biochim Biophys Res Commun, 2002,295(2):532-9
[36] Scheller J,Rose-John S.Interleukin-6 and its receptor:from bench to bedside[J].Med Microbiol Immunol,2006,195(4):173-183.
[37] Picanco-Castro V,Fontes AM,Heinz S,et al.The chimeric cytokine Hyper-IL-6 enhances the efficiency of lentiviral gene transfer in hepatocytes both in vitro and in vivo[J].Biotechnol Lett,2008,30(2):215-220.
[1] Ankoma-sey V.Hepatic regeneration-revisiting the myth of Prometheu- s.News Physiol Sci,1999,14:149-155.
[2] Taub R.Liver regeneration:from myth to mechanism.Nat Rev Mol Cell Biol,2004,5(10):836-847.
[3] Kitamura T,Watanabe S,Sato N.Liver regeneration,liver cancers and cyclins.J Gastroenterol Hepatol,1998,13(Supp1):96-99.
[4] Fischer M,Goldschmitt J,Peschel C,et al.A bioactive designer cytokine for human hepatopoietic progenitor cell expansion. Nat Biotechnol,1997,15(2):142-145.
[5] Zvibel I,Brill S,Kariv R,et al.Chimeric molecule IL-6/soluble recept- or is a potent mitogen for fetal hepatocytes.J Cell Physiol,2004, 200 (2):245-252
[6] Scheller J,Rose-John S.Interleukin-6 and its receptor :from bench to bedside.Med Microbiol Immunol,2006,(Epub ahead of print)
[7] Okaya A,Kitanaka J,Kanaka N,et al.Oncostatin M inhibits proliferation of oval cells,OC15-5,including differentiation into hepatocytes.Am J Pathol,2005,166(3):709-719.
[8] Nakamura K,Nonaka H,Saito H,et a1.Hepatoyte proliferation and tissue remodeling is impaired after liver injury in oncostatin M receptor knockout mice.Hepatology,2004,39(3):635-644.
[9] Rahmani M,Peron P,Weitzman J,et a1.Functional cooperation between JunD and NF-kappaB in rat hepatocytes.Oncogene,2001,20(37):5132-5142.
[10] Laurent S,Horsmans Y,Starkel P,et al.Disrupted NF-kappaB activation after partial hepatectomy dose not impair hepatocyte proliferation in rats .World J Gastroenterol,2005,11(46):7345-7350.
[11] Debidda M,Wang L,Zang H,et al.A role of STAT3 in Rho GTPase-regulated cell migration and proliferation.J Biol Chem 2005,280(17): 17275- 17285.
[12] Streetz KL,Luedde T,Manns MP,et a1.Interleukin 6 and liver regener- ation.Gut,2000,47(2):309-312.
[13] Galun E,Zeira E,Pappo O,et a1.Liver regeneration induced by a designer human IL-6/sIL-6R fusion protein reverse severe hepatocel- lular injury.FASEB J,2000,14(13):1979-1987.
[14]Taub R ,Greenbaum LE,Peng Y,Transcriptional regulatory signals define cytokine-dependent and -independent pathways in liver regeneration.Semin Liver Dis,1999,19(2):117-127.
[15]DeAngelis RA,Markiewski MM,Lambris JD.Liver regeneration:a link to inflammation through complement.Adv Exp Med Biol,2006,586:17-34.
[16]Strey CW,Markiewski M,Mastellos D,et al.The proinflammatory mediators C3a and C5a are essential for liver regeneration.J Exp Med,2003,198 (6):913-923
[17]Fausto N.Liver regeneration.J Hepatol,2000,32(1 Suppl):19-31.
[2] Fausto NJ.Liver regeneration[J].Hepatol,2000,32:19-31.
[3] Fischer M, Goldschmitt J, Peschel C,et al. A bioactive designer cytokine for human hemotopoietic progenitor cell expansion[J].Nature Biotechnol,1997,15:142-145.
[4] Galun E, Axelrod JH. The role of cytokines in liver failure and regeneration: potential new therapies. Biochim Biophys Acta, 2002,1592(3):354-358.
[5]郭国宁,周跃,张正丰.成骨细胞特异性转录因子 Cbfa1 重组腺病毒的构建、纯 化及表达[J].免疫学杂志.2005,21(2):150-154.
[6] D.L.斯配克特,R.D.戈德曼,L.A.莱因万德.细胞试验指南[M].黄培堂等译. 北京: 科学出版社,2001:830-831.
[7] 董世武,应大君,刘光久,等.小鼠 Osf2/Cbfal 基因的重组腺病毒构建及鉴定 [J ]. 第三军医大学学报. 2003, 25 (21) : 1947—1954.
[8] K.K.杰恩, 基因治疗学[M].任斌等译.西安:世界图书出版西安公司,2000: 603
[9] Kinoshita K,limuro Y,Fujimoto J,et a1.Targeted and regulable expression of transgenes in hepatic stellate cells and myofibroblasts in culture and in vivo using an adenoviral Cre /loxP system to antagonise hepatic fibrosis[J].Gut,2007,56(3):396 -404.
[10] Liu L,Pedersen E F. Partial abstracts of the 3rd annual meeting of the China RNA society & 1st international RNA workshop in China:SESSION1:mRNA processing biochemical identification of nuclear protein p30 specifically interacting with upstrea- m repressi[J]. 生物化学与生物物理学报(英文版),2004,36(2):147
[11] Kalyuzhniy O,Di Paolo NC,Si lvestry M,et al.Adenovirus serotype 5 hexon is critic- al for virus infection of hepatocytes in vivo[J]. Proc Natl Acad Sci U S A.2008,105 (14):5483-5488.
[12] Robert JJ,Gauffeny I,Maccario J,et al.Degenerated pIX-IVa2adenoviral vectorsequences lowers reacquisition of the E1genes duringvirus amplification in293cells[J].Gene Ther,2001,8(22):1713-1720.
[13]Crouzet J,Naudin L,Orsini c,et a1.Recombinational construction in escherichia coli of infectious adenoviral genomes[J].ProcAcad Sci USA.1997,94(4):1414—1419.
[14]Davis AR,Wivel NA,Palladino JL,et a1.Construction of adenoviral vectors [J].Mol Biotechnol,2001,18(1):63—70.
[15] He TC, Zhou S, da Costa LT, et al. A simplified system for generating recombinant adenoviruses[J]. Proc Natl Acad Sci U S A. 1998 ,95(5):2509-14
[16]CONG Tie-chuan,LU Zhe-ming,Li Yong,et al.High efficient generation of replication-defective adenoviruses containing thymidine kinase by homogeneous recombination in bacteria[J].Chin Med . 2007,120(18):1622-1625.
[17]吕慧,赵蔚明,于修平,等. E 型沙眼衣原体 MOMP 基因重组腺病毒的构建及免疫原性研究[J].山东大学学报(医学版),2006,44(12):1189-1193.
[18]李玉峰,姜平,蒋文明,等.猪繁殖与呼吸综合征病毒 GP5 蛋白重组腺病毒的构建与免疫原性测定[J].中国病毒学,2006,21(4):364-367.
[19]Kalyanasundaram S,Feinstein S,Nicholson JP, et al. Coacervate microspheres as carriers of recombinant adenoviruses[J].Cancer Gene Ther, 1999, 6(2): 107-112.
[20]中华医学会传染病与寄生虫病学分会、肝病学分会联合修订.病毒性肝炎防治方案[J].中华肝脏病杂志,2000,8(6):324-329.
[21] Zimmers TA,Pierce RH,Mckillop IH,et al. Resolving the role of IL-6 in liver regeneration [J].Hepatol,2003,38:1590-1591.
[22] Boulanger MJ,Chow D,Brevnova EE,et al.Hexameric structure and assembly of the interleukin-6/IL-6a-receptor/gp130 complex[J].Science,2003,300:2101-2104.
[23] Oberg HH,Wesch D,Grussel S,et al. Differential expression of CD126 and CD130 mediates different STAT-3 phosphorylation in CD4+CD25- and CD25high regulatory T cells[J].Int Immunol,2006,18(4):555-563.
[24] Igaz P,Horvath A,Horvath B,et al. Soluble interleukin-6 receptor (sIL-6R) makes IL-6R negative T cell line respond to IL-6;it inhibits TNF production[J].ImmunolLetter,2000,71:143-148.
[25]Jones SA,Richards PJ,Scheller J,et al.IL-6 transsignaling:the in vivo consequences[J].Interferon Cytokine Res 2005(25):241-253.
[26] Zvibel I, Brill S, Kevital R, et al. Chimeric molecule IL-6/soluble receptor is a potent mitogen for fetal hepatocytes[J] .Cell Physiol,2004,200:245-252.
[27] Peters M, Solem F, Schirmacher P, et al. IL-6 and soluble IL-6R induce stem cell factor (SCF) and Flt-3 ligand expression in vivo and in vitro[J]. Exp Hematol, 2001, 29:146–155.
[28] Peters M, Blinn G, Jostock T,et al. Combined interleukin 6 and soluble interleukin 6 receptor accelerates murine liver regeneration[J].Gastroenterol,2000,119(6):1163 –1171.
[29] Male Peters,Guido Blinn,Fian Solem,et al.In vivo and in vitro activities of the gp130-stimulating designer cytokine Hyper-IL-6[J]. Immunol,1998,161:3575-3581.
[30] Yeoh GC,Ernst M,Rose-J0hn S,Akhurst B,et al.Opposing roles of gp130-mediated STAT-3and ERK-1/2signaling in liver progenitor cell migration and proliferation[J].Hepatoology,2007,45(2):486-494.
[31] Richards PJ,Nowell MA,Horiuchi S,et al.Functional characterization of a soluble gp130 isform and its therapeutic capacity in an experimental model of inflammatory arthritis[J].Arthritis Rheum,2006,54(5):1662-1672.
[32] Hecht N,Pappo O,Shouval D,et al.Hyper-IL-6 gene therapy reverses fulminant hepatic failure[J].Mol Therap ,2001,3:683-687.
[33]Gotze KS, Keller U, Rose-John S, et al. Gp130-stimulating designer cytokine hyper-interleukin-6 synergizes with murine stroma for long-term survival of primitive human hematopietic progenitor cells[J]. Exp Hematol, 2001,29(7): 822-32
[34] Wang XP, Schunck M, Kallen KJ, et al. The interleukin-6 cytokine system regulates epidermal permeability barrier homeostasis[J]. J Invest Dermatol.2004 Jul;123(1):124-31
[35] Sun Y, Marz P, Otten U,et al. The effect of gp130-stimulation on glutamate-induced excitotoxicity in primary hippocampal neurons[J]. Biochim Biophys Res Commun, 2002,295(2):532-9
[36] Scheller J,Rose-John S.Interleukin-6 and its receptor:from bench to bedside[J].Med Microbiol Immunol,2006,195(4):173-183.
[37] Picanco-Castro V,Fontes AM,Heinz S,et al.The chimeric cytokine Hyper-IL-6 enhances the efficiency of lentiviral gene transfer in hepatocytes both in vitro and in vivo[J].Biotechnol Lett,2008,30(2):215-220.
[1] Ankoma-sey V.Hepatic regeneration-revisiting the myth of Prometheu- s.News Physiol Sci,1999,14:149-155.
[2] Taub R.Liver regeneration:from myth to mechanism.Nat Rev Mol Cell Biol,2004,5(10):836-847.
[3] Kitamura T,Watanabe S,Sato N.Liver regeneration,liver cancers and cyclins.J Gastroenterol Hepatol,1998,13(Supp1):96-99.
[4] Fischer M,Goldschmitt J,Peschel C,et al.A bioactive designer cytokine for human hepatopoietic progenitor cell expansion. Nat Biotechnol,1997,15(2):142-145.
[5] Zvibel I,Brill S,Kariv R,et al.Chimeric molecule IL-6/soluble recept- or is a potent mitogen for fetal hepatocytes.J Cell Physiol,2004, 200 (2):245-252
[6] Scheller J,Rose-John S.Interleukin-6 and its receptor :from bench to bedside.Med Microbiol Immunol,2006,(Epub ahead of print)
[7] Okaya A,Kitanaka J,Kanaka N,et al.Oncostatin M inhibits proliferation of oval cells,OC15-5,including differentiation into hepatocytes.Am J Pathol,2005,166(3):709-719.
[8] Nakamura K,Nonaka H,Saito H,et a1.Hepatoyte proliferation and tissue remodeling is impaired after liver injury in oncostatin M receptor knockout mice.Hepatology,2004,39(3):635-644.
[9] Rahmani M,Peron P,Weitzman J,et a1.Functional cooperation between JunD and NF-kappaB in rat hepatocytes.Oncogene,2001,20(37):5132-5142.
[10] Laurent S,Horsmans Y,Starkel P,et al.Disrupted NF-kappaB activation after partial hepatectomy dose not impair hepatocyte proliferation in rats .World J Gastroenterol,2005,11(46):7345-7350.
[11] Debidda M,Wang L,Zang H,et al.A role of STAT3 in Rho GTPase-regulated cell migration and proliferation.J Biol Chem 2005,280(17): 17275- 17285.
[12] Streetz KL,Luedde T,Manns MP,et a1.Interleukin 6 and liver regener- ation.Gut,2000,47(2):309-312.
[13] Galun E,Zeira E,Pappo O,et a1.Liver regeneration induced by a designer human IL-6/sIL-6R fusion protein reverse severe hepatocel- lular injury.FASEB J,2000,14(13):1979-1987.
[14]Taub R ,Greenbaum LE,Peng Y,Transcriptional regulatory signals define cytokine-dependent and -independent pathways in liver regeneration.Semin Liver Dis,1999,19(2):117-127.
[15]DeAngelis RA,Markiewski MM,Lambris JD.Liver regeneration:a link to inflammation through complement.Adv Exp Med Biol,2006,586:17-34.
[16]Strey CW,Markiewski M,Mastellos D,et al.The proinflammatory mediators C3a and C5a are essential for liver regeneration.J Exp Med,2003,198 (6):913-923
[17]Fausto N.Liver regeneration.J Hepatol,2000,32(1 Suppl):19-31.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |