溃结安对大鼠IL-6基因mRNA水平的影响及pSilencer干扰载体的构建
|
摘要
目的:研究溃结安对溃疡性结肠炎(Ulcerative colitis,UC)大鼠白细胞介素-6(interleukin 6,IL-6)基因mRNA水平的影响及构建具有特异性阻断大鼠IL-6基因的小干扰RNA(small intefferening RNA,siRNA)表达体系,为确定溃疡性结肠炎发病基因及研究维药西帕依溃结安治疗溃疡性结肠炎作用机理提供新的方法。方法:Wistar大鼠分为正常组,溃疡性结肠炎模型组,生理盐水阴性对照组组,5-氨基水杨酸阳性对照组,溃结安干预组共5组,用2,4-二硝基氯苯(DNCB)复合乙酸方法造成溃疡性结肠炎大鼠模型,从结肠中提取总RNA经逆转录-聚合酶链反应(RT-PCR)得到目的基因的cDNA,经PCR得到目的基因的扩增片段。应用siRNA设计软件在大鼠IL-6的mRNA上寻找特异性的短核苷酸序列并设计合成两条互补的小发夹结构的DNA序列,经退火后形成双链DNA片段,采用基因克隆技术,将其克隆到pSilencer~(TM)1.0-U6的载体中,转化DH5α大肠杆菌,提取质粒,酶切方法和测序法对重组体进行鉴定。结果:溃结安降低了大鼠IL-6基因mRNA水平。酶切和序列测定表明,成功构建了大鼠IL-6基因的siRNA表达体系。结论:大鼠IL-6基因表达的变化可能与溃疡性结肠炎的发生发展相关联;溃结安治疗作用可能与其下调了IL-6基因表达有关;成功构建siRNA表达体系,为确定溃疡性结肠炎发病基因及研究维药溃结安治疗溃疡性结肠炎的靶点打下了基础。
Objective:To investigate the expression of intedeukin 6(IL-6)in intestinal tissue of rat with Ulcerative Colitis(UC)and further studies the pathogenesis of UC.To construct the specific small interfereing RNA(siRNA)expression vector that can block the rat IL-6 gene by RNA interference(RNAi)technique,which provides a new approach to determine the onset gene of Ulcerative Colitis and investigate the target of the Uygur medicine Kuijie'an(KJA)that can treat the UC.Methods:Wistar rats were derided into 5 groups:control group,UC groups,NS group,5-ASA group,UC rats induced by DNCB and acetic acid.Observe and assay the expression of IL-6 gene by RT-PCR.According to the encoding sequence of mRNA of rat IL-6 provided by Genbank,two pieces of specific hairpin structure short chain oligonucleotide sequences were designed and synthesized by using the siRNA software.The annealed oligonucleotide fragment were subcloned into pSilencer ~(TM)1.0-U6 expression vector by gene clone technique and transformed into DH5αE.coli.Finally the plasmid evaluated by using enzyme cutting and sequencing. Results:Expression of IL-6 gene remarkably increased in intastinal tissue of rats with UC as compared with control group;but it remarkably decreased after theraphy of KJA. Identification using enzyme cutting and sequencing showed that the IL-6 siRNA expression vector was constructed successfully.Conclusion:It demonstrated tentavtively that KJA had an effect on UC.Successful construction of rat IL 6 siRNA expression vector will facilitate further studies of determine the onset gene of Ulcerative Colitis and investigate the target of the Uygur medicine KJA that can treat.the Ulcerative Colitis.
Objective:To investigate the expression of intedeukin 6(IL-6)in intestinal tissue of rat with Ulcerative Colitis(UC)and further studies the pathogenesis of UC.To construct the specific small interfereing RNA(siRNA)expression vector that can block the rat IL-6 gene by RNA interference(RNAi)technique,which provides a new approach to determine the onset gene of Ulcerative Colitis and investigate the target of the Uygur medicine Kuijie'an(KJA)that can treat the UC.Methods:Wistar rats were derided into 5 groups:control group,UC groups,NS group,5-ASA group,UC rats induced by DNCB and acetic acid.Observe and assay the expression of IL-6 gene by RT-PCR.According to the encoding sequence of mRNA of rat IL-6 provided by Genbank,two pieces of specific hairpin structure short chain oligonucleotide sequences were designed and synthesized by using the siRNA software.The annealed oligonucleotide fragment were subcloned into pSilencer ~(TM)1.0-U6 expression vector by gene clone technique and transformed into DH5αE.coli.Finally the plasmid evaluated by using enzyme cutting and sequencing. Results:Expression of IL-6 gene remarkably increased in intastinal tissue of rats with UC as compared with control group;but it remarkably decreased after theraphy of KJA. Identification using enzyme cutting and sequencing showed that the IL-6 siRNA expression vector was constructed successfully.Conclusion:It demonstrated tentavtively that KJA had an effect on UC.Successful construction of rat IL 6 siRNA expression vector will facilitate further studies of determine the onset gene of Ulcerative Colitis and investigate the target of the Uygur medicine KJA that can treat.the Ulcerative Colitis.
引文
铩颷1]Jiang XL,Cui HF.An analysis of 10218 ulcerative colitis cases in China[J].World JGastroenterol,2002,8(1):158-161.
[2]Bartnik W.Inflammatory bowel disease Polish contribution[J].J Physiol Pharmacol,2003,54(3):205-210
[3]Kusugami K,Fukasu A,Tanimoto M,et al.Elevation ofinterleukin-6 in inflammatorybowel disease is macrophage and epithelial cell- dependent[J].Dig Dis Sci,1995,40(5):949.
[4]Toyoda H,Yang H,Rotter JI,et al.Ulcerative colitis:a genetically heterogeneousdisorder defined by genetic(HLA class II)and subclinical(antineutrophilcytoplasmic antibodies)markers[J].J Cli Invest,1993,92(2):1080-1084
[5]Jesus K,Yamamoto-Furusho,Luis F,et al.Clinical and Genetic Heterogeneity inMexican Patients With Ulcerative Colitis[J].Human Immunology,2003,64(1):119-123
[6]Hideki Masuda,Yasuo Takahashi,Satoshi Asai.Distinct Gene Expression ofOsteopontin in Patients With Ulcerative Colitis[J].Journal of Surgical Research,2003,111(1):85-90
[7]Hiroaki N,Nagamu I,Toshifumi H.Association between the interleukin-1 receptorantagonist polymorphism and Ulcerative Colitis with younger age at diagnosis[J].Immunology Letters,2003,90(1):53-57
[8]Juan Luis Fdez-Morera,Luis Rodrigo,Antonio Lopez-Vazquez.MHCCLASSIChain-Related Gene A Transmembrane Polymorphism Modulates theExtension of Ulerative Colitis[J].Human Immunology,2003,64(1):816-822
[9]MacDonald TT,Monteleone G;Pender SL.Recent developmentsin the immunologyof inflammatory bowel disease[J].Scand J Immunol,2000,51(1):229.
[10]Papadakis KA,Targan SR.Role of cytokines in the pathogenesis of inflammatorybowel disease[J].A nnu Rev Med,2000,51:2892298.
[11]Ardizzone S,Porro GB.Inflammatory bowel disease:new in2sights into pathogenesisand treatment[J].J Int Med,2002,252:4752496.
[12]王伟宁,张熙纯,刘丽.活动期溃疡性结肠炎发病机制的免疫学探讨.中国现代医学杂志2003;13:74-78
[13]Sartor RB.Cytokine in intestinal inflammation:Pathophysiological and clinicalconsiderations[J].Gastroenterology,1994,106(2):533-539
铩颷14]Hyams J S,Fitzgerald J E,Treem WR et al.Relationship of functional and antigenicinterleukin 6 to disease activity in inflammatory bowel disease[J].Gastroenterology,1993,104(5):1285-1292
[15]Holtkamp W,Stollberg T,Reis HE.Serum interleukin-6 is.relatedto disease activitybut not disease specificity in inflammatory bowel disease[J].J Clin Gastroenterol,1995,20(2):123
[16]Holub MC,Mako E,Devay T,et al.Increased interleukin-6 levels,interleukin-6receptor and gpl30 expression in peripheral lymphocytes of patients withinflammatory bowel disease[J].Scand J Gastroenterol,1998,228:47-50
[17]Grottrup-Wolfers E,Moeller J,Karbach U,et al.Elevated cell-associated levels ofinterleukin 1βand interleukin 6 in inflamed mucosa of inflammatory bowel disease[J].Eur J Cli Invest,1996,26(2):115-122
[18]Kazuo K,Atsushi F,Mitsune T,et al.Elevation of interleukin-6 in inflammatorybowel disease is macrophage and epithelial cell- dependent[J].Dig Dis Sci,1995,40(5):949
[19]崔淑兰,汤斌,梁坚,等.溃疡性结肠炎患者血清IL-6测定及临床意义探讨[J].医学综述,1998,4(5)248.
[20]李琪佳,宫恩聪,刘叔平,等.溃疡性结肠炎发病机制的免疫病理学及分子病理学研究[J].中华消化杂志,2000,20(5):324-326
[21]郭海建,邓长生,夏冰.溃疡性结肠炎患者白细胞介素-6活性研究[J].中华消化杂志,2001,21(4):223-225
[22]丁伟群,林庚金,徐三荣,等.溃疡性结肠炎发病中白介素水平的变化[J].复旦学报(医学版),2001,28(4)330-335
[23]卢艺涛,高静,姚桂琴.溃疡性结肠炎患者相关细胞因子的实验性研究[J].现代预防医学,2005,32(7):735-736
[24]Hammond SM,Bemstain E,Beach D,et al.An RNA-directed muciease mediate posttranscriptional gene silencing in Drosophila cells[J].Nature,2000,404(6775):293-296.
[25]Zamore PD,Thschl T,Sharp PA,et al.RNAi:Double-stranded RNA directs theATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals[J].cell,2000,101(1):25-33.
[26]Scherr M,Battmer K,Winkler T,et al.Specific inhibition of ber-abl gene expressionby small interfering RNA[J].Blood,2003,101(4):1566-1569.
[27]胡福泉主编.现代基因操作技术(第一版)[M].北京:人民军医出版社.
铩颷28]卢圣栋主编.现代分子生物学技术(第二版)[M].北京:协和医科大学出版社.
[29]陈昭,王洪艳,张新刚等.溃疡性结肠炎动物模型的实验研究.北华大学学报(自然科学版).2005,6(1):65-67
[30]李林,王竹立,柯剑婷,等.实验性溃疡性结肠炎动物模型选择.世界华人消化杂志,2001,9:5842585.
[31]张文明.脾虚型溃疡性结肠炎患者症状观察与形态学研究[J].中医药学刊,2005,23(8):1391-1393
[32]Atreya R,Neurath MF.Involvement of IL-6 in the pathogenesis of inflammatorybowel disease and colon cancer.Clin Rev Allergy Immuno12005;28:187-196
[33]Kitamura K,Nakamoto Y,Kaneko S,Mukaida N.Pivotal roles of interleukin-6 intransmural inflammation in murine T cell transfer colitis.J Leukoe Biol2004;76:1111-1117
[34]邓长生.溃疡性结肠炎患者白细胞介素-6活性研究.中华消化杂志2001;21:223-225
[35]Raddatz D,Bockemuhl M,Ramadod G.Quantitative measurement of eytokinemRNA in inflammatory bowel disease:relation to clinical and endoscopic activity andoutcome.Eur J Gastroenterol Hepatol 2005;17:547-5571
[36]Street ME,de'Angelis G,Camaeho-Hubner C,Criovannelli G,Ziveri MA,BaeehiniPL,Bemaseoni S,Sansebastiano G,Savage MO.Relationships between serum IGF-1,IGFBP-2,interleukin-lbeta and interleukin-6 in inflammatory bowel disease.HormRes 2004;61:159-164
[37]贾百灵,侯晓华,白细胞介素-6与溃疡性结肠炎的关系胃肠病学和肝病学杂志2004;13:220-221
铩颷1]Kombluth A,Sachar DB.Ulcerative colitis practice guidelines in adults.Americancollege of gastroenterology,practice parameters committee.Am J Gastroenterol1997;92:204-211
[2]Eaden J,Abrams K,Ekbom A,Jackson E,Mayberry J.Colorectal cancer preventionin ulcerative colitis:a case- control study.Aliment Pharmacol Ther 2000;14:145153
[3]喻德洪.《现代肛肠外科学》.第1版.北京:人民军医出版社,1997:287-292
[4]Fonager K,Sorensen HT,Olsen J.Change in incidence of Crohn's disease andulcerative colitis in Denmark.A study based on the national registry of patients,1981-1992.hat J Epidemiol 1997;26:1003-1008
[5]Russel MG.Changes in the incidence of inflammatory bowel disease:what does itmean? Eur J Intern Med 2000;11:191-196
[6]Jiang XL,Cui HF.An analysis of 10218 ulcerative.colitis cases in China.World JGastroentero12002;8:158-161
[7]陈灏珠,丁训节,廖履坦,杨秉辉,翁心华.《实用内科学》.第11版.北京:人民卫生出版社,2001:1785-1789
[8]Campieri M,Gionchetti E Bacteria as the cause of ulcerative colitis.Gut2001;48:132135
[9]Nakamura RM,Barry M.Serologic markers in inflammatory bowel disease(IBD).Med Lab Obs 2001;33:8-15
[10]Eggena M,Cohavy O,Parseghian MH,Hamkalo BA,Clemens D,Targan SR,Gordon LK,Braun J.Identification of histone HI as a cognate antigen of theulcerative colitis-associated marker antibody pANCA.J Autoimmun 2000;14:83-97
[11]Gordon LK,Eggena M,Targan SR,Braun J.Mast cell and neuroendocrinecytoplasmic autoantigen(s)detected by monoclonal pANCA antibodies.ClinImmunol 2000;94:42-50
[12]MacDonald NJ,Shivers WY,Narum DL,Plum SM,Wingard JN,Fuhrmann SR,Liang H,Holland-Linn J,Chen DH,Sim BK.Endostatin binds tropomyosin.Apotential modulator of the antitumor activity of endostatin.J Biol Chem2001;276:25190-25196
[13]Taniguchi M,Geng X,Glazier KD,Dasgupta A,Lin JJ,Das KM.Cellular immuneresponse against tropomyosin isoform 5 in ulcerative colitis.Clin Immunol2001;101:289-295
铩颷14]Schreibcr S.Experimental immunomodulatory therapy of inflammatory boweldisease.Netherl J Mcd 1998;53:24-31
[15]李琪佳,徐敏,宫恩聪.细胞因子与溃疡性结肠炎.中国煤炭工业医学杂志2001;4:411-413
[16]Stevens C,Walz G,Singaram C,Lipman ML,Zanker B,Muggia A,Antonioli D,Pe-ppercorn MA,.Strom TB.Tumor necrosis factor-α interleukin-1β,and interleukin-6expression in inflammatory bowel disease.Dig Dis Sei,1992,37:818-826
[17]Breese EJ,Michie CA,Mieholls W,Nieholls SW,Murch SH,Williams CB,DomizioP,Walker-Smith JA,Macdonald TT.Tumor necrosis factor alpha-producing cells inthe intestinal mucosa of children with inflammatory bowel disease.Gastroenterology,1994;106:1455-1466
[18]丁伟群,林庚金,徐三荣,钱立平.溃疡性结肠炎发病中白介素水平的变化.复旦学报2001;7:330-335
[19]Almawi WY,Lipman ML,Stevens AC,Zanker B,Hadro ET,Strom TB.Abrogationof glucoeortieoid-mediated inhibition of T cell proliferation by the synergistic of IL-1,IL-6 and TNF-γ.J Immunol 1991;146:3523-3527
[20]Willems J,Joniau M,Cinque S,van Damme J.Human granuloeyte chemotacticpeptide(IL-8)as a specific neutrophil degranulator:comparison with other monokines.Immunology 1989;67:540-542
[21]Mazzucchelli L,Hauser C,Zgraggen K,Wagner H,Hess M,Laissae JA,Mueller C.Expression of interleukin-8 gene in inflammatory bowel disease is related to thehistological grade of active inflammation.Am J Pathol 1994;144:997-1007
[21]Nielsen OH,Koppen T,Rudiger N,Horn Y,Eriksen J,Kirman L.Involvement ofinterleukin-4 and 10 in inflammatory bowel disease.Dig Dis Sci 1996;41:1786-1793
[22]Tagore A,Gonsalkorale WM,Praviea V,Hajeer AH,McMahon R,Whorwell P J,Sinnott PJ,Hutchinson IV.Interleukin-10(IL-10)genotypes in inflammatory boweldisease.Tissue Antigen 1999;54:386-390
[23]Jijon HB,Churchill T,Malfair D,Wessler A,Jewell LD,Parsons HG,Madsen KL.Inhibition of poly(ADP-ribose)polymerase attenuates inflammation in a model ofchronic colitis.Am J Physiol Gastrointest Liver Physio12000;279:641-651
[24]Sartor RB.Pathogenesis and immune mechanisms of chronic inflammatory boweldiseases.Am J Gastroenterol 1997;92(12 Suppl):5-11
[25]Neurath MF,Finotto S,Fuss I,Boirivant M,Galle PR,Strober W.Regulation ofT-cell apoptosis in inflammatory bowel disease:to die or not to die,that is the mucosal question.Trends Immunol 2001;22:21-26
铩颷26]Targan SR,Deem R,Liu M,Wang S,Nel A.Definition of a lamina propria T-cellresponsive state:Enhanced cytokine responsiveness of T-cells stimulated through theCD2 pathway.J Immtmol 1995;154:664-675
[27]Wills- Karp M,Santeliz J,Karp C1.The germless theory of allergicdisease:revisitingthe hypothesis.Nature Rev,2001,1:69-75.
[28]Hugot JP,Chamaillard M,Zouali H,et al.Association of NOD21eucine- rich repeatvariants with susceptibility to Crohn's disease.Nature,2001,411:599-603.
[29]Duerr RB,Barmada MU,Zhang L,et al.High density genome scanin Crohn diseaseshows confirmed linkage to chromosone 14q11-2=12.Am J Hum Genet,2000,66:1257-1262.
[30]Stokkers PCF,Reitsma PH,Tytgat GNJ,et al.HLA- DR and - DQphenotypes ininflammatory bowel disease:a meta - analysis.Gut,1999,45:395-401.
[31]Orchard TR,Thiyagaraja S,Welsh KI,et al.Clinical phenotype isrelated to HLAgenotype in the peripheral arthropathies of inflammatorybowel disease.Gastroenterology,2000,118:274-278.
[32]朱元民,王勤河,刘玉兰,等.溃疡性结肠炎环境因素致病作用的研究[J].临床内科杂志,2002,19(5):350-351.
[33]CAMPIERI M,GIONCHETTIP.Bacteria as the cause of ulcerative colites[J].Gut,2001,48:132-135.
[34]徐三荣.吸烟与溃疡性结肠炎[J].国外医学·生理、病理科学与临床分册,1998,18(4):379-380.
[35]刘凤芹,楚更五,李子华,等.心理因素与溃疡性结肠炎[J].健康心理学杂志,2001,9(4):307.
[36]王英,陈言东,堂彤,等.溃疡性结肠炎危险因素的病例对照研究[J].中华消化杂志,2004,24(5):305.
[37]郑家驹.炎症性肠病[M].上海:上海科学技术文献出版社,1998.16-21.
[38]Miller M J,Angeles FM,Reuter BK,Bobrowski P,Sandoval M.Dietary antioxidantsprotect gut epithelial cells from oxidant-induced apoptosis.BMC Complement AltemMed2001;1:11
[39]Yoshida Y,Iwai A,Itoh K,Tanaka M,Kato S,Hokari R,Miyahara T,Koyama H,Miura S,Kobayashi M.Role of inducible nitric oxide synthase in dextran sulphatesodium-induced colitis.Aliment Pharmacol Ther 2000;14(Suppl 1):26-32
[40]Grisham MB.Oxidants and free radicals in inflammatory bowel disease.Lancet 1994;344:859-861
铩颷41]Cortield AP,Myerscough N,Longman R,Sylvester P,Arul S,Pignatelli M.Mucinsand mucosal protection in the gastrointestinal tract:new prospects for mucins in thepathology of gastrointestinal disease.Gut 2000;47:589-594
[42]Campbell B J,Rowe GE,Leiper K,Rhodes JM.Increasing the intra-Golgi pH ofcultured LS 174T goblet-differentiated cells mimics the decreased muein sulfation andincreased Thomsen-Fdedenreich antigen(Gal betal-3GalNac alpha-)expression seenin colon cancer.Glycobiology 2001;11:385-393
[43]Einerhand AW,Renes IB,Makkink MK,Van Der Sluis M,Buller HA,Dekker J.Roleof mucins in inflammatory bowel disease:important lessons from experimentalmodels.Eur J Gastroenterol Hepatol
[44]徐萍,周小江,吕农华等.溃疡性结肠炎组织中环氧化酶-2于一氧化氮合酶的表达及意义.中华消化内镜杂志,2004,1.
[2]Bartnik W.Inflammatory bowel disease Polish contribution[J].J Physiol Pharmacol,2003,54(3):205-210
[3]Kusugami K,Fukasu A,Tanimoto M,et al.Elevation ofinterleukin-6 in inflammatorybowel disease is macrophage and epithelial cell- dependent[J].Dig Dis Sci,1995,40(5):949.
[4]Toyoda H,Yang H,Rotter JI,et al.Ulcerative colitis:a genetically heterogeneousdisorder defined by genetic(HLA class II)and subclinical(antineutrophilcytoplasmic antibodies)markers[J].J Cli Invest,1993,92(2):1080-1084
[5]Jesus K,Yamamoto-Furusho,Luis F,et al.Clinical and Genetic Heterogeneity inMexican Patients With Ulcerative Colitis[J].Human Immunology,2003,64(1):119-123
[6]Hideki Masuda,Yasuo Takahashi,Satoshi Asai.Distinct Gene Expression ofOsteopontin in Patients With Ulcerative Colitis[J].Journal of Surgical Research,2003,111(1):85-90
[7]Hiroaki N,Nagamu I,Toshifumi H.Association between the interleukin-1 receptorantagonist polymorphism and Ulcerative Colitis with younger age at diagnosis[J].Immunology Letters,2003,90(1):53-57
[8]Juan Luis Fdez-Morera,Luis Rodrigo,Antonio Lopez-Vazquez.MHCCLASSIChain-Related Gene A Transmembrane Polymorphism Modulates theExtension of Ulerative Colitis[J].Human Immunology,2003,64(1):816-822
[9]MacDonald TT,Monteleone G;Pender SL.Recent developmentsin the immunologyof inflammatory bowel disease[J].Scand J Immunol,2000,51(1):229.
[10]Papadakis KA,Targan SR.Role of cytokines in the pathogenesis of inflammatorybowel disease[J].A nnu Rev Med,2000,51:2892298.
[11]Ardizzone S,Porro GB.Inflammatory bowel disease:new in2sights into pathogenesisand treatment[J].J Int Med,2002,252:4752496.
[12]王伟宁,张熙纯,刘丽.活动期溃疡性结肠炎发病机制的免疫学探讨.中国现代医学杂志2003;13:74-78
[13]Sartor RB.Cytokine in intestinal inflammation:Pathophysiological and clinicalconsiderations[J].Gastroenterology,1994,106(2):533-539
铩颷14]Hyams J S,Fitzgerald J E,Treem WR et al.Relationship of functional and antigenicinterleukin 6 to disease activity in inflammatory bowel disease[J].Gastroenterology,1993,104(5):1285-1292
[15]Holtkamp W,Stollberg T,Reis HE.Serum interleukin-6 is.relatedto disease activitybut not disease specificity in inflammatory bowel disease[J].J Clin Gastroenterol,1995,20(2):123
[16]Holub MC,Mako E,Devay T,et al.Increased interleukin-6 levels,interleukin-6receptor and gpl30 expression in peripheral lymphocytes of patients withinflammatory bowel disease[J].Scand J Gastroenterol,1998,228:47-50
[17]Grottrup-Wolfers E,Moeller J,Karbach U,et al.Elevated cell-associated levels ofinterleukin 1βand interleukin 6 in inflamed mucosa of inflammatory bowel disease[J].Eur J Cli Invest,1996,26(2):115-122
[18]Kazuo K,Atsushi F,Mitsune T,et al.Elevation of interleukin-6 in inflammatorybowel disease is macrophage and epithelial cell- dependent[J].Dig Dis Sci,1995,40(5):949
[19]崔淑兰,汤斌,梁坚,等.溃疡性结肠炎患者血清IL-6测定及临床意义探讨[J].医学综述,1998,4(5)248.
[20]李琪佳,宫恩聪,刘叔平,等.溃疡性结肠炎发病机制的免疫病理学及分子病理学研究[J].中华消化杂志,2000,20(5):324-326
[21]郭海建,邓长生,夏冰.溃疡性结肠炎患者白细胞介素-6活性研究[J].中华消化杂志,2001,21(4):223-225
[22]丁伟群,林庚金,徐三荣,等.溃疡性结肠炎发病中白介素水平的变化[J].复旦学报(医学版),2001,28(4)330-335
[23]卢艺涛,高静,姚桂琴.溃疡性结肠炎患者相关细胞因子的实验性研究[J].现代预防医学,2005,32(7):735-736
[24]Hammond SM,Bemstain E,Beach D,et al.An RNA-directed muciease mediate posttranscriptional gene silencing in Drosophila cells[J].Nature,2000,404(6775):293-296.
[25]Zamore PD,Thschl T,Sharp PA,et al.RNAi:Double-stranded RNA directs theATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals[J].cell,2000,101(1):25-33.
[26]Scherr M,Battmer K,Winkler T,et al.Specific inhibition of ber-abl gene expressionby small interfering RNA[J].Blood,2003,101(4):1566-1569.
[27]胡福泉主编.现代基因操作技术(第一版)[M].北京:人民军医出版社.
铩颷28]卢圣栋主编.现代分子生物学技术(第二版)[M].北京:协和医科大学出版社.
[29]陈昭,王洪艳,张新刚等.溃疡性结肠炎动物模型的实验研究.北华大学学报(自然科学版).2005,6(1):65-67
[30]李林,王竹立,柯剑婷,等.实验性溃疡性结肠炎动物模型选择.世界华人消化杂志,2001,9:5842585.
[31]张文明.脾虚型溃疡性结肠炎患者症状观察与形态学研究[J].中医药学刊,2005,23(8):1391-1393
[32]Atreya R,Neurath MF.Involvement of IL-6 in the pathogenesis of inflammatorybowel disease and colon cancer.Clin Rev Allergy Immuno12005;28:187-196
[33]Kitamura K,Nakamoto Y,Kaneko S,Mukaida N.Pivotal roles of interleukin-6 intransmural inflammation in murine T cell transfer colitis.J Leukoe Biol2004;76:1111-1117
[34]邓长生.溃疡性结肠炎患者白细胞介素-6活性研究.中华消化杂志2001;21:223-225
[35]Raddatz D,Bockemuhl M,Ramadod G.Quantitative measurement of eytokinemRNA in inflammatory bowel disease:relation to clinical and endoscopic activity andoutcome.Eur J Gastroenterol Hepatol 2005;17:547-5571
[36]Street ME,de'Angelis G,Camaeho-Hubner C,Criovannelli G,Ziveri MA,BaeehiniPL,Bemaseoni S,Sansebastiano G,Savage MO.Relationships between serum IGF-1,IGFBP-2,interleukin-lbeta and interleukin-6 in inflammatory bowel disease.HormRes 2004;61:159-164
[37]贾百灵,侯晓华,白细胞介素-6与溃疡性结肠炎的关系胃肠病学和肝病学杂志2004;13:220-221
铩颷1]Kombluth A,Sachar DB.Ulcerative colitis practice guidelines in adults.Americancollege of gastroenterology,practice parameters committee.Am J Gastroenterol1997;92:204-211
[2]Eaden J,Abrams K,Ekbom A,Jackson E,Mayberry J.Colorectal cancer preventionin ulcerative colitis:a case- control study.Aliment Pharmacol Ther 2000;14:145153
[3]喻德洪.《现代肛肠外科学》.第1版.北京:人民军医出版社,1997:287-292
[4]Fonager K,Sorensen HT,Olsen J.Change in incidence of Crohn's disease andulcerative colitis in Denmark.A study based on the national registry of patients,1981-1992.hat J Epidemiol 1997;26:1003-1008
[5]Russel MG.Changes in the incidence of inflammatory bowel disease:what does itmean? Eur J Intern Med 2000;11:191-196
[6]Jiang XL,Cui HF.An analysis of 10218 ulcerative.colitis cases in China.World JGastroentero12002;8:158-161
[7]陈灏珠,丁训节,廖履坦,杨秉辉,翁心华.《实用内科学》.第11版.北京:人民卫生出版社,2001:1785-1789
[8]Campieri M,Gionchetti E Bacteria as the cause of ulcerative colitis.Gut2001;48:132135
[9]Nakamura RM,Barry M.Serologic markers in inflammatory bowel disease(IBD).Med Lab Obs 2001;33:8-15
[10]Eggena M,Cohavy O,Parseghian MH,Hamkalo BA,Clemens D,Targan SR,Gordon LK,Braun J.Identification of histone HI as a cognate antigen of theulcerative colitis-associated marker antibody pANCA.J Autoimmun 2000;14:83-97
[11]Gordon LK,Eggena M,Targan SR,Braun J.Mast cell and neuroendocrinecytoplasmic autoantigen(s)detected by monoclonal pANCA antibodies.ClinImmunol 2000;94:42-50
[12]MacDonald NJ,Shivers WY,Narum DL,Plum SM,Wingard JN,Fuhrmann SR,Liang H,Holland-Linn J,Chen DH,Sim BK.Endostatin binds tropomyosin.Apotential modulator of the antitumor activity of endostatin.J Biol Chem2001;276:25190-25196
[13]Taniguchi M,Geng X,Glazier KD,Dasgupta A,Lin JJ,Das KM.Cellular immuneresponse against tropomyosin isoform 5 in ulcerative colitis.Clin Immunol2001;101:289-295
铩颷14]Schreibcr S.Experimental immunomodulatory therapy of inflammatory boweldisease.Netherl J Mcd 1998;53:24-31
[15]李琪佳,徐敏,宫恩聪.细胞因子与溃疡性结肠炎.中国煤炭工业医学杂志2001;4:411-413
[16]Stevens C,Walz G,Singaram C,Lipman ML,Zanker B,Muggia A,Antonioli D,Pe-ppercorn MA,.Strom TB.Tumor necrosis factor-α interleukin-1β,and interleukin-6expression in inflammatory bowel disease.Dig Dis Sei,1992,37:818-826
[17]Breese EJ,Michie CA,Mieholls W,Nieholls SW,Murch SH,Williams CB,DomizioP,Walker-Smith JA,Macdonald TT.Tumor necrosis factor alpha-producing cells inthe intestinal mucosa of children with inflammatory bowel disease.Gastroenterology,1994;106:1455-1466
[18]丁伟群,林庚金,徐三荣,钱立平.溃疡性结肠炎发病中白介素水平的变化.复旦学报2001;7:330-335
[19]Almawi WY,Lipman ML,Stevens AC,Zanker B,Hadro ET,Strom TB.Abrogationof glucoeortieoid-mediated inhibition of T cell proliferation by the synergistic of IL-1,IL-6 and TNF-γ.J Immunol 1991;146:3523-3527
[20]Willems J,Joniau M,Cinque S,van Damme J.Human granuloeyte chemotacticpeptide(IL-8)as a specific neutrophil degranulator:comparison with other monokines.Immunology 1989;67:540-542
[21]Mazzucchelli L,Hauser C,Zgraggen K,Wagner H,Hess M,Laissae JA,Mueller C.Expression of interleukin-8 gene in inflammatory bowel disease is related to thehistological grade of active inflammation.Am J Pathol 1994;144:997-1007
[21]Nielsen OH,Koppen T,Rudiger N,Horn Y,Eriksen J,Kirman L.Involvement ofinterleukin-4 and 10 in inflammatory bowel disease.Dig Dis Sci 1996;41:1786-1793
[22]Tagore A,Gonsalkorale WM,Praviea V,Hajeer AH,McMahon R,Whorwell P J,Sinnott PJ,Hutchinson IV.Interleukin-10(IL-10)genotypes in inflammatory boweldisease.Tissue Antigen 1999;54:386-390
[23]Jijon HB,Churchill T,Malfair D,Wessler A,Jewell LD,Parsons HG,Madsen KL.Inhibition of poly(ADP-ribose)polymerase attenuates inflammation in a model ofchronic colitis.Am J Physiol Gastrointest Liver Physio12000;279:641-651
[24]Sartor RB.Pathogenesis and immune mechanisms of chronic inflammatory boweldiseases.Am J Gastroenterol 1997;92(12 Suppl):5-11
[25]Neurath MF,Finotto S,Fuss I,Boirivant M,Galle PR,Strober W.Regulation ofT-cell apoptosis in inflammatory bowel disease:to die or not to die,that is the mucosal question.Trends Immunol 2001;22:21-26
铩颷26]Targan SR,Deem R,Liu M,Wang S,Nel A.Definition of a lamina propria T-cellresponsive state:Enhanced cytokine responsiveness of T-cells stimulated through theCD2 pathway.J Immtmol 1995;154:664-675
[27]Wills- Karp M,Santeliz J,Karp C1.The germless theory of allergicdisease:revisitingthe hypothesis.Nature Rev,2001,1:69-75.
[28]Hugot JP,Chamaillard M,Zouali H,et al.Association of NOD21eucine- rich repeatvariants with susceptibility to Crohn's disease.Nature,2001,411:599-603.
[29]Duerr RB,Barmada MU,Zhang L,et al.High density genome scanin Crohn diseaseshows confirmed linkage to chromosone 14q11-2=12.Am J Hum Genet,2000,66:1257-1262.
[30]Stokkers PCF,Reitsma PH,Tytgat GNJ,et al.HLA- DR and - DQphenotypes ininflammatory bowel disease:a meta - analysis.Gut,1999,45:395-401.
[31]Orchard TR,Thiyagaraja S,Welsh KI,et al.Clinical phenotype isrelated to HLAgenotype in the peripheral arthropathies of inflammatorybowel disease.Gastroenterology,2000,118:274-278.
[32]朱元民,王勤河,刘玉兰,等.溃疡性结肠炎环境因素致病作用的研究[J].临床内科杂志,2002,19(5):350-351.
[33]CAMPIERI M,GIONCHETTIP.Bacteria as the cause of ulcerative colites[J].Gut,2001,48:132-135.
[34]徐三荣.吸烟与溃疡性结肠炎[J].国外医学·生理、病理科学与临床分册,1998,18(4):379-380.
[35]刘凤芹,楚更五,李子华,等.心理因素与溃疡性结肠炎[J].健康心理学杂志,2001,9(4):307.
[36]王英,陈言东,堂彤,等.溃疡性结肠炎危险因素的病例对照研究[J].中华消化杂志,2004,24(5):305.
[37]郑家驹.炎症性肠病[M].上海:上海科学技术文献出版社,1998.16-21.
[38]Miller M J,Angeles FM,Reuter BK,Bobrowski P,Sandoval M.Dietary antioxidantsprotect gut epithelial cells from oxidant-induced apoptosis.BMC Complement AltemMed2001;1:11
[39]Yoshida Y,Iwai A,Itoh K,Tanaka M,Kato S,Hokari R,Miyahara T,Koyama H,Miura S,Kobayashi M.Role of inducible nitric oxide synthase in dextran sulphatesodium-induced colitis.Aliment Pharmacol Ther 2000;14(Suppl 1):26-32
[40]Grisham MB.Oxidants and free radicals in inflammatory bowel disease.Lancet 1994;344:859-861
铩颷41]Cortield AP,Myerscough N,Longman R,Sylvester P,Arul S,Pignatelli M.Mucinsand mucosal protection in the gastrointestinal tract:new prospects for mucins in thepathology of gastrointestinal disease.Gut 2000;47:589-594
[42]Campbell B J,Rowe GE,Leiper K,Rhodes JM.Increasing the intra-Golgi pH ofcultured LS 174T goblet-differentiated cells mimics the decreased muein sulfation andincreased Thomsen-Fdedenreich antigen(Gal betal-3GalNac alpha-)expression seenin colon cancer.Glycobiology 2001;11:385-393
[43]Einerhand AW,Renes IB,Makkink MK,Van Der Sluis M,Buller HA,Dekker J.Roleof mucins in inflammatory bowel disease:important lessons from experimentalmodels.Eur J Gastroenterol Hepatol
[44]徐萍,周小江,吕农华等.溃疡性结肠炎组织中环氧化酶-2于一氧化氮合酶的表达及意义.中华消化内镜杂志,2004,1.