紫癜圣愈散对CITP模型小鼠外周血共刺激分子CD86影响的实验研究
详细信息    本馆镜像全文|  推荐本文 |  |   获取CNKI官网全文
摘要
目的:通过观察应用紫癜圣愈散后CITP模型小鼠外周血中血小板计数及共刺激分子CD86水平的变化,初步探讨共刺激分子CD86与CITP发病机制的关系及紫癜圣愈散对CITP的作用机理。
     方法:采用注射豚鼠抗小鼠血小板血清(APS)的免疫造模方法,建立BALB/c小鼠CITP模型;将BALB/c小鼠60只随机分为6组:正常组、模型组、激素对照组、紫癜圣愈散低、中、高剂量组;应用紫癜圣愈散对模型小鼠进行实验研究,进行实验观察:(1)全自动血细胞计数仪检测CITP模型小鼠血小板计数;(2)酶联免疫吸附法(enzyme-linkedimmunosorbent assay,ELISA)测定CITP模型小鼠外周血CD86的水平;(3)观察各组小鼠造模及用药前后毛色、活动、大便、反应、出血点和瘀斑等一般情况的变化。
     结果:①造模后小鼠的血小板减少,模型对照组与正常对照组相比,有显著差异(P<0.05),模型对照组与正常对照组相比CD86水平明显升高,有显著差异(P<0.05),初步证实了模型复制成功;②紫癜圣愈散低、中、高剂量组、激素组均能提高模型小鼠PLT计数,激素组、紫癜圣愈散中、高剂量组两两比较(P>0.05)无显著性差异;紫癜圣愈散低剂量组与激素组比较(P<0.05)有显著性差异;③紫癜圣愈散低、中、高剂量组、激素组均能降低模型小鼠CD86水平,激素组、紫癜圣愈散中、高剂量组两两比较(P>0.05)无显著性差异;紫癜圣愈散低剂量组与激素组比较(P<0.05),有显著性差异;④各治疗组的小鼠一般状态均较模型对照组好,进食量、活动量增加,出血现象减轻。
     结论:紫癜圣愈散能升高模型小鼠外周血小板数,降低CITP模型小鼠外周血中CD86水平,对BALB/c小鼠CITP动物模型具有较好的治疗效果。
Objective:To investigate the mechanism by which Zidian Shengyu Pulvis(ZSP)affects the development of CITP through observing changes of PLT number and the peripheral blood expression of CD86 in CITP model mice.
     Methods:1.The mice were divided into six groups:normal,model,hormone control,low dose of ZSP,middle dose of ZSP,and high dose of ZSP.2.The chronic ITP model was established by intraperitoneally injection of anti-BALB/c platelet serum(APS).3.The platelet number in mice was calculated by an automatic blood cell counter.The peripheral blood expression of CD86 in mice was measured by ELISA.The mice in treatment groups were gavaged with ZSP for 14 consecutive days starting 8 hours agter the 1st abdominal cavity injection of APS.
     Results:The PLT number in mice of the model group was lower than that in normal group,and the expression of CD86 in mice of the model group was statistically higher than that in the normal group(P<0.05).Compared with the model group,mice in all the treatment groups showed statistically higher PLT number and lower CD86 expression levels(P<0.05).No significant difference was observed among the middle dose of ZSP,high dose of ZSP and the hormone group.
     Conclusion:CD86 was expressed in peripheral blood at higher level in the ITP model mice.ZSP has functions of lowering peripheral blood expression of CD86 and increasing PLT number.It plays a role in the treatment of ITP by regulating immunological functions to reduce PLT destruction.
引文
[1]侯丕华.原发性血小板减少性紫癜的中医论治体会[J].中日友好医院学报2002,(1):59.
    [2]王延丰.生血消斑饮治疗特发性血小板减少性紫癜临床观察[J].天津中医,2001,(4):3.
    [3]何文清.加减归脾汤治疗特发性血小板减少性紫癜疗效观察[J].辽宁中医杂志,2006,33(9):1146.
    [4]周永明,程军.原发性血小板减少性紫癜的病机特点和治疗对策[J].辽宁中医杂志,2001,(12):714.
    [5]李海燕.紫癜冲剂治疗免疫性血小板减少性紫癜的临床研究[J].辽宁中医杂志,2002,(1):33.
    [6]唐旭东,麻柔.特发性血小板减少性紫癜的中医病机和治疗[J]内科急危重症杂志,2006,12(6):262-263.
    [7]叶明.辨证治疗原发性血小板减少性紫癜32例疗效观察[J].北京中医,2002,21(3):141-142.
    [8]李文梅,赵保丽.活血化瘀治疗原发性血小板减少性紫癜[J].黑龙江中医药,2000,(1):10-11.
    [9]潘铭.辨证治疗血小板减少性紫癜123例分析[J].甘肃中医,2001.(6):31.
    [10]赵海东.益气生血汤治疗特发性血小板减少性紫癜45例[J].山西中医,2005,21(4):17-18.
    [11]周清华,魏自敏.消斑饮口服液治疗慢性原发性血小板减少性紫癜100例[J].实用中医内科杂志,2004,(8):2.
    [12]周全良.陈学达运用柴平生血汤治疗特发性血小板减少性紫癜经验[J].中医药临床杂志,2006,18(3):219-220.
    [13]刘安平,赵萍,陈志雄,张惠臣.紫癜灵治疗慢性特发性血小板减少性紫癜的临床观察[J],湖北中医杂志 2005,27(9):13-14.
    [14]庄万传,李秀梅,韩秀华,朱贵华.雷公藤多苷联合长春地辛治疗难治性ITP 的临床研究[J].江苏医药,2006,32(2):184-185.
    [15]赵仕伟,莫激勤,陈元标等.紫癜汤治疗原发性血小板减少性紫癜疗效观察[J].实用中医内科杂志,2005,19(4):358-359.
    [16]吴顺杰,梁冰,李达.梁冰老中医治疗慢性特发性血小板减少性紫癜的经验研究[J].时珍国医国药,2007,18(9):2270-2271.
    [17]徐瑞荣,李丙,刘朝霞,曹芳.中西医结合治疗难治性特发性血小板减少性紫癜疗效观察[J].中国中医急症,2004,13(1):19-20.
    [18]封青海.贞芪扶正胶囊治疗慢性血小板减少性紫32例[J].陕西中医,2005,26(12):1312-1313.
    [19]Yang R,Han ZC.Pathogenesis and management of chronic idiopathic thrombocytopenic purpura:an update.Int J Hetnatol,2000,71:18.
    [20]Cines DB,Blanchette VS.Tmmune thrombocytopenic purpura.NEngl J Med,2002,346:995.
    [21]Zhou B,Zhao H,Yang RC,et al.Multi-dysfunctional pathophysiology in ITP.Crit Rev in Oncol/Hematol,2005,54:107.
    [22]Zandman-Goddard G,Shoenfel Y.HIV and autoimmunity.Autoimmunity Reviews,2002,1:329.
    [23]乔丽津.病毒感染与特发性血小板减少性紫癜.中国小儿血液,2001,(6):6.
    [24]Belshe RB Text book of Human Virology Littleton:Massachusetts PSG publishing Company,1984,912.
    [25]Crapnell K In vitro infection of megakaryocytes and their precursors by human cytomegalorious Blood,2000,95:485.
    [26]李兰娜,王玉琼.儿童慢性特发性血小板减少性紫癜与巨细胞病毒感染的关系[J].中国全科医学,2003,(6):10.
    [27]林雯,肖燕等.人巨细胞病毒感染致血小板减少性紫癜病因研究[J].中华微生物学和免疫学杂志,2002,(22):5.
    [28]Gasbarrini A,Franceschi F,Tartaglione R,et al.Regression of autoimmune thrombocytopenia after eradication of Helicobacter pylori.Lancet,1998,352:878.
    [29]Stasi R,Rossi Z,Stipa E,et al.Helicobacter pylori eradication in the management of patients with idiopathic thromboeytopenic purpura,Am J Med,2005,118:414.
    [30]Yoshikawa T,Morooka M,Suga S,et al Five cases of thrombocy topenia induced by primary human herpesvims 6 infection[J]Acta pacdiatr Jpn,1998,40:278-281.
    [31]马晓彤,吕冬梅等.人类第6型疱疹病毒与特发性血小板减少性紫癜[J].中华血液学杂志,2000,21(3):135-137.
    [32]董学君,刘忠民等.特发性血小板减少性紫癫与人类疱疹病毒6型感染关系初探[J].肿瘤瘤防治杂志,2003,(10):1.
    [33]严媚,古丽娜·沙丁,顾立达.人类微小病毒B19与小儿血液病的关系[[J].新疆医科大学学报,2003,26(5).
    [34]李爱敏,张继红等.人类细小病毒B19感染与小儿急性特发性血小板减少性紫癜[J].小儿急救医学,2003,(10):5.
    [35]曾佑群,刘瑜,张振琪.人细小病毒B19感染与儿童血液疾病的关系[J]医技杂志,2003(10):6.
    [36]Heegaard ED,Rostho JS,Petersen BL,et al.Role of parvovirus B19 infection in Childhood idiopathic thrombocytopenic purpura[J].ActaPaediatr,1999,88(6):614-617.
    [37]Kooter AJ,Van derlinden PW,De Klerk C.et al.Acute idiopathic thrombocytopenic purpura in adults following viral infection:report of two cases[J].Neth J Med,2002,60(4):174.
    [38]孙凤兰.疫苗接种后血.小板减少性紫[J].中国小儿血液,2002,(7):4.
    [39]Stanworth SJ,Turner DM,Brown J,et al.Major histocompatibility complex susceptibility genes and immune thrombocytopenic purpura in Caucasian adults.Hematology,2002,7:119.
    [40]Nomura S,Matsuzaki T,Ozaki Y,et al.Clinical significance of HLA-DRB1·0410in Japanese patients with idiopathic thrombocytvpenic purpura.Blood,1998,91:3616.
    [41]Foster CB,Zhu S,Erichsen HC,et al.Polymorphisms in inflammatory cytokines and Fcgamma receptors in childhood chronic immune thrombocytopenic purpura;a pilot study.Br J Haematol,2001,113:596.
    [42]Satoh T,Pandey JP,Okazaki Y,et al.Single nucleotide polymorphisms of the inflammatory cytokine genes in adults with chronic immune thrombocytopenic purpura. Br J Haematol, 2004,124:796.
    
    [43] Sanchez Margalet V, Martin-Romerv C, Santos-Alvarez J, et al. Role of leptin as an immune-omodulator of blood mononuclear cells: mechanisms of action. Clin Exp Immunol, 2003,133:11.
    
    [44] Zhan M, Zhao H, Yang R,et al. Serum leptin levels in patients with idiopathic thrombocytopenic purpura. Eur J Haematol, 2004,72:348.
    
    [45] Ren H, Zhao H, Wang T, et al. Leptin enhances in vitro secretion of IgG antiplatelet antibodies by splenocytes and peripheral blood mononuclear cells from patients with chronic idiopathic thrombocytopenic purpura. Clin Immunol, 2006,120:205.
    
    [46] Sanna V, Di Giacomo A, La Cava A, et al. Leptin surge precedes onset of autoimmune encephalomyelitis and correlates with development of pathogenic T cell responses. J Clin Invest, 2003, 111;241.
    
    [47] Martin-Romero C, Santos-Alvarez J, Gobema R, et al. human leptin enhances activation and proliferation of human circulating T lymphocytes. Cell Immunol, 2000,199:15.
    
    [48] Zhou B, Zhao H, Yang RC, et al. Multi-dysfunctional pathophysiology in ITP. Crit Rev in Oncol/Haematol,2005, 54:107.
    
    [49] Kuwana M, Kaburaki J, Kitasato H,et al. Immunodominant epitopes on glycoprotein IIb-IIIa recognized by autoreactive T cells in patients with immune thrombocytopenic purpura. Blood, 2001,98:130.
    
    [50] Wardemann H, Yurasov S, Schaefer A, et al. Predominant autoantibody production by early human B cell precursors. Sciettce, 2003,301:1374.
    
    [51] Takahashi R, Sekine N, Nakatake T. Influence of monoclonal antiplatelet glycoprotein antibodies on in vitro human megakaryocyte colony formation and proplatelet formation. Blood, 1999, 93 :1951.
    
    [52] Chang M, Nakagawa PA, Williams SA, et al. Immune thrombocytopenic purpura plasma and purified ITP monoclonal autoantibodies inhibit megakaryocytopoiesis in vitro. Blood, 2003, 102:887.
    
    [53] McMillan R, Wang L, Tourer A, et al. Suppression of in vitro megakaryocyte production by anti-platelet autoantibodies from adult patients with chronic ITP.Blood, 2004,103:1364.
    
    [54] Houwerzijl EJ, Blom NR, van der Want JJ, et al. Ultrastructural study shows morphologic features of apoptosis and para-apoptosis in megakaryocytes from patients with idiopathic thrombocytopenic purpura. Blood, 2004,103:500.
    
    [55] Binstadt BA, Geha RS, Bonilla FA. IgG Fc receptor polymorphisms in human disease; implications for intravenous immunoglobulin therapy. J Allergy Clin Immunol,2003,111:697.
    
    [56] Carcao MD, Blanchette VS, Wakefield CD, et al. Fcgamma receptor IIa and IIIa polymorphisms in childhood immune thrombocytopenic purpura. Br J Haematol,2003,120:135.
    
    [57] Webber NP, Mascarenhas JO, Crow MK, et al. Funtional properties of lymphocytes in idiopathic thromlpcytopenic purpura [J]. Hum Immunol, 2001,62:1346-1355.
    
    [58] Olsson B, Andersson PO, Jernas M, et al. T-cell-mediated cytotoxicity toward platelets in chronic idiopathic thrombocytopenic purpura. Nat Med,2003,9:1123-1124.
    
    [59] Wang T, Zhao H, Ren H, et al. Type 1 and type 2 T-cell profiles in idiopathic thrombocytopenic purpura. Haematologica, 2005,90:914.
    
    [60] Freeman GJ,Gribben JG,Boussiotis VA,et al. Cloning of B7-2: a CTLA-4 counter-receptor that costimulates human T cell proliferation. Science. 1993 ,262 :909-11.
    
    [61] Cross AH,Girard TJ,Giacoletto KS,et al. Long-term inhibition of murine experimental autoimmune encephalomyelitis using CTLA-4-Fc supports a key role for CD28 costimulation. J Clin Invest. 1995 Jun;95(6):2783-9.
    
    [62] Djukanovic R.The role of co-stimulation in airway inflammation.Clin Exp Allergy. 2000 Jun;30 Suppl 1:46-50.
    
    [63] Hernandez HJ,Sharpe AH,Stadecker MJ,et al. Experimental murine schistosomiasis in the absence of B7 costimulatory molecules: reversal of elicited T cell cytokine profile and partial inhibition of egg granuloma formation.J Immunol. 1999 Mar 1;162(5):2884-9.
    
    [64] Linsley RS, Clark EA, Ledbetter JA. T-cell antigen CD28 mediates adhesion with B cells by interacting with activation antigen B7/BB-1.Proc Natl Acad Sci USA.1990 Jul;87(13):5031-5.
    [65]Lazetie S,Leong SR,Chang JC,et al.Chimeric co-stimulatory molecules that selectively act through CD28 or CTLA-4 on human T cells.J Biol Chem.2002 Oct 11;277(41):38660-8.
    [66]Jeannin P,Magistrelli G,Aubry JP,et al.Soluble CD86 is a costimulatory molecule for human T lymphocytes.Immunity.2000 Sep;13(3):303-12.
    [67]Henn V,Slupsky,JR,Grafe M,et al.CD40 ligand on activated platelets triggers an inflammatory reaction of endothelial cells Nature,1998;391(6667):591-594.
    [68]Nagahama M,Nomura S,Kanazawa S,et al.Signigieanee of chemokines and solubleCD40 ligand patients with autoimmune thromboeytopenic purpura.Eur J Haematol,2002,69:303-308.
    [69]赵艳霞,高清平,陈友华等.特发性血小板减少性紫癜患者外周血淋巴细胞协同刺激分子的表达.中华血液学杂志,2003,24:474-476.
    [70]Hou M,Peng J,Shi Y,et al.Mycophenolatemofetil(MMF)for the treatment of steroid-resistant idiopathic thrombocytopenic purpura Eur J Haematol,2003,70:353-357.
    [71]Peng J,Liu C,Liu D,et al.Effeets of B7-bloeking agent and/or CsA on induction of platelet-speeific T-cell anergy in chronic autoimmune thrombocytopenic purpura Blood,2003,101:2721-2726.
    [72]YOSHMURA C,NOMURA S,NAGAHAMA M,et al.Plasma-soluble Fas(APO-1,CD95)and soluble Fas ligand in immune thromboeytopenic purpura[J].Eur J Haematol,2000,64(4):219-224.
    [73]侯明.成人特发性血小板减少性紫癜的规范化诊断与治疗[J].临床血液学杂志,2008,21(1):3-4.
    [74]赵艳红,周晋.特发性血小板减少性紫癜患者糖皮质激素受体的表达[J].基础医学与临床,2005,25(4):362-365.
    [75]王学文.成人慢性免疫性血小板减少性紫癜的治疗进展[J].医学研究生学报,2003,16(1):51-53.
    [76]British Committee for Standards in Haematology General Haematology Task Force Guidelines for the investigation and management of idiopathic thrombocytopenic purpura in adults,children and in pregnancy.Br J Haetnatol,2003,120:574.
    [77]李艳.成人特发性血小板减少性紫癜现代诊断和治疗[J].中华实用内科杂志,2005,6(25):568-571.
    [78]段婷,常瑛,刘锦丽.环孢素A治疗难治性特发性血小板减少性紫癜的近期疗效观察[J].中国内科杂志,2000,39(9):630.
    [79]魏立,孔佩艳,严巨明.环孢素A联合肾上腺皮质激素治疗难治性ITP的疗效观察[J].西南国防医药,2004,2(14):158-160.
    [80]黄国辉,林世光,张瑞官.干扰素治疗小儿慢性特发性血小板减少性紫癜15例[J].中国小儿血液,2005,4(10):182.
    [81]Stasi R,Pagano A,Stipa E,et al.Rituximab chimeric antiCD20 monoclonal antibody traetment for adults with chronic idiopathic thrombocytopenic purpura.Blood,2001,98:952.
    [82]Stasi R,Stipa E,Forte V,et al.Variable patterns of response to rituximab treatment in adults with chronic idiopathic thrombocytopenic purpura.Blood,2002,99:3872.
    [83]张永利,候芸华.来氟米特治疗慢性特发性血小板减少性紫癜的疗效研究[J].医师进修杂志,2005,9(28):48-49.
    [84]陈协群,白庆咸,王一苇,等.重组人血小板生成素治疗特发性和血液肿瘤化疗后血小板减少症的Ⅲ期临床研究[J].第四军医大学学报,2004,2(3):261-264.
    [85]白洁,邵宗鸿.第八次全国血液学学术会议纪要[J].中国实用内科杂志,2005,6(25):572-573.
    [86]庞丽萍,杨保青,孟庆祥,等,特发性血小板减少性紫癜的病因及疗效探讨[J].临床血液学杂志,2005,1,(18):11-13.
    [87]Emilia G,Longo G,Luppoi M,etal.Helicobacter pylori eradication can induce platelet recovery in idiopathic thrombocytopenic purpura[J].Blood,2001,97:812-814.
    [88]姜玉华,陈学良,程玉峰,等.脾脏放射治疗老年人难治性原发性血小板减少性紫癜17例报告[J].东大学学报,2005,4(43):367-368.
    [89]任宝志,韩磊,冯长伟,等.三维适形脾区放疗与药物治疗慢性特发性血小板 减少性紫癜疗效比较[J].临床血液学杂志,2005,5(18):279-280.
    [90]叶斌,张世珍.雷公藤多甙治疗难治性血小板减少性紫癜20例[J].浙江临床医学,2000,2(1):54.
    [91]陈永平,苟正英.火把花根联合强的松治疗难治性免疫性血小板减少性紫癜(附27例报告)[J].广西医学,2002,24(3):409-410.
    [92]郭良耀,徐朝焰,赵小亭CVP方案治疗慢性特发性难治性血小板减少性紫癜的临床疗效观察[J].新医学,2002,33(5):298.
    [93]American Society of Hematology 41 annual meeting.New Orleans,1999,94(10pt 1):646.
    [94]Carpenter D,Yee T.Megakaryocytopenia inW/W v mice is accompanied by an increase in size within ploidy groups and acceleratation of maturation.Blood,1989,74:94-98.
    [96]Ebbe S,Lebin J,Miller K,et al.Thrombocytopoietic response to immunothrombocutopenia in nude mice.Blood,1987,69:192-198.
    [96]Teeling TL,Jansen-Hendriks T,Kuijpers TW,et al.Therapeutic efficacy of intravenous immunoglobulin preparations depends on the immunoglobulin G diners studies in experimental immune thrombocytopenia.Blood,2001,98:1095-1099.
    [97]Hansen RJ,Baldiasar JP.Pharmacokinetics pharmacodynamics and platelet binding of an anti-glycoprotein Ⅱb/Ⅲa Monoclonal Antibody(7E3)in the rat a quantitative rat model of immune thrombocytopenic purpura J Pharmacol Exp Ther,2001,298:165-171.
    [98]Hansen RJ,Baldiasar JP.Effects of intravenous immunoglobulin on platelet count and antiplatelet antibody disposition in a rat model of immune thrombocytopenia.Blood,2002,100:2087-2093.
    [99]Crow AR,Lazarus AH.Role of Fcgamma receptors in the pathogenesis and treatment of idiopathic thrombocytopenic purpura Jpediatr Hematol Oncol.2003,25(Suppl 1):S14-18.
    [100]Coopers,Heddle NM,HaasM,et al.Intravenous(Ⅳ)anti-D and Ⅳimmunoglobulin achieve acute platelet increases by different mechanism,modulation of cytokine and platelet responses toⅣ anti-D by FcgammaRⅡa and FcganmaRⅢa polymorphisns Br,J Haematol.2004,124:511-518.
    [101]Song S,Crow AR,Siragam V,et al.Monoclonal antibodies that minic the action of anti-D in the amelioration of murine ITP act by a mechanisn distinct from that of ⅣⅠg.Blood,2005,105:1546-1548.
    [102]Carcao MD,B lanchette VS,Wakefield CD,et al.Fcgamma reeeptorⅡa and Ⅲa polymorphisms in childhood immune thrombocytopenic purpura Br J Haematol.2003,120:135-141.
    [103]Crow AR,Song S,Semple JW,et al.ⅣⅠg inhibits reticuloendothelial system function and ameliorates murine passive-immune thrombocytopenia independent of anti-idiotype reactivity.Br JHaemato,2001,115:679-686.
    [104]McKenzie SE.Humanized mouse model of FoR clearance in immune platelet disorders.Blood Rew,2002,16:3-5.
    [105]张爱军.免疫性血小板减少性紫癜动物模型的研究进展[J].中国实验血液学杂志,2006,14(3):623-626.
    [106]杨宇飞.免疫性血小板减少性紫癜动物模型的建立[J].中华血液学杂志,1994,15(3):16.
    [107]Blazevic V,Trubey CM,Shearer GM.Analysis of the costimulatory requirements for generating human virus-specific in vitro T helper and effector responses.J Clin Immunol.2001 Jul;21(4):293-302.
    [108]Lanier LL,O'Fallon S,Somoza C,et aLCD80(B7)and CD86(B70)provide similar costimulatory signals for T cell proliferation,cytokine production,and generation of CTL.J Immunol.1995 Jan 1;154(1):97-105.
    [109]Azuma M,Yssel H,Phillips JH,et al.Functional expression of B7/BB1 on activated T lymphocytes.J Exp Med.1993 Mar 1;177(3):845-50.
    [110]Punt JA,Osborne BA,Takahama Y,et al.Negative selection of CD4~+CD8~+thymocytes by T cell receptor-induced apoptosisrequires a costimulatory signal that can be provided by CD28.JExp Med.1994 Feb 1;179(2):709-13.
    [111]曾艳,高清平,陈友华,等.特发性血小板减少性紫癜患者外周血中树突状细胞共刺激分子表达研究.临床内科杂志,2005,22(3):206.
    [112]赵艳霞,高清平,陈友华,等.特发性血小板减少性紫癜患者外周血淋巴细胞协 同刺激分子的表达。中华血液学杂志,2003,24(9):474-475.
    [113]张春梅,赵春亭,滕清良,等.ITP患者外周血淋巴细胞共刺激分子表达及意义.[J]山东医药,2004,44(16):6-7.

© 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