摘要
炎症性肠病(IBD)是一类重要的肠道疾病,临床症状多变,治疗效果差,易复发,其病因和发病机制到目前为止仍不是十分明确。我们从神经-内分泌-免疫网络与机体免疫稳态调节的角度出发,探讨IL-10在IBD发生、发展中的作用,以期为探讨IBD的发生、发展及其与神经-内分泌-免疫网络的关系奠定基础。本实验以建立炎症性肠病大鼠动物模型为出发点,并且分别于造模后第3天、7天、14天、21天、28天取大鼠脑组织、结肠组织和血液。结合炎症性肠病各时期病理组织学变化,利用免疫组化方法研究脑组织、结肠组织中IL-10表达的变化,利用ELISA方法检测血清中IL-10浓度的变化。结果表明,随着肠管病理变化的加剧,IL-10在大脑皮层和下丘脑的表达升高,而在结肠组织中的表达和在血清中的浓度逐渐降低;随着肠管病理变化的减轻,IL-10在大脑皮层和下丘脑的表达逐渐降低,同时,在结肠组织中的表达和血清中的浓度逐渐升高并趋向于正常。提示脑组织中IL-10的变化与IBD的发生、发展呈正相关;而结肠组织和血清中IL-10的变化与IBD的发生、发展呈负相关。研究结果提示,IL-10作为神经-内分泌-免疫网络中共用的信息分子可能在IBD的发生、发展中起着重要作用。
Inflammatory bowel disease (IBD) is an important intestinal disease, including the crohn disease (CD) and ulcerative colitis (UC).There is a great diversity of clinical symptom, a bad therapeutic efficacy and an easy recidivism. Especially in Europe and the America, it gets a high morbility, simultaneously, the morbility is increasing gradually in recent years. The etiopathogenisis and nosogenesis are unclear now. Most researchers think they are related with infection, immunologic abnormality, heredity and so on. Immunologic abnormality is considered to play the most important role among all of these factors.T cells play an extremely important role in the immune system. According to the difference of cytokines that CD4 +T cell secretes and mediating immune function, they are divided into Th1 and Th2, Th1 cells express IFN-γand IL-2 mainly, and Th2 cells express IL-4, IL-5 and IL-10 etc. More and more evidences prove that Th1 and Th2 have different roles in the course of mediating immune response. Th1/Th2 imbalance has been considered to be one of the pathogenesis of IBD. When Th1 or Th2 cells enhance mucosa inflammatory response, the amount of the suppression cells, regulatory T cells and the secretion of cytokines decreased significantly, which demonstrated in the development of inflammatory bowel disease the imbalance plays an important role. In short, both T cells and their sub-type (Th1,Th2,Th3) and cytokines (pro-inflammatory cytokines, anti-inflammatory cytokines and growth factor) play a significant role in the pathogenesis course of IBD immunity.
Nervous, endocrine and immune form a complex network, named neuro-endocrine-immune regulatory network, which controls the body's various vital movement, maintains homeostasis of the body. When the homeostasis is disrupted, certain parts in the network will respond and do some change in the function, meanwhile, some change occurs in the referred network, leading to certain diseases and the emergence of pathological process. Neuro-endocrine-immune disorder is a common feature of all the diseases. All the clinical diseases have a nerve-endocrine-immune disorder. The disease process is actually a kind of disorder for homeostasis regulation of nerve-endocrine-immune network, however, among the nerve, endocrine and immune disorders, immune disorder is principal. Since the occurrence and development of IBD are closely related to T cells and their cytokines, the nerve - endocrine - immune regulatory network plays an important role in regulating T-cell-mediated immunity. However, whether the nerve-endocrine-immune regulatory network mediates the development and occurrence of IBD, few are reported today.
In view of the above, the experiment is to use artificially induced IBD rats as models, make use of immunohistochemical method, to discuss the effect and mechanism of IL-10 in the occurrence and development of IBD, from the view of the nerve-endocrine-immune network and the immune homeostasis regulation. It will lay a strong foundation to ensure the relationship of the course of IBD and neuroendocrine immune network, provide a new way to prevent and treat the inflammatory bowel disease (IBD) , and offer experimental basis for further study at nerve endocrine immune network theory. Therefore, we make IBD models by giving an enema with trinitro-benzene-sulfonic acid. Put the IBD rats to death on the 3rd ,7th ,14th ,21st, 28th day. Take the brain and colon tissues. Detect the expression level of IL-10 in the intestinal canal and brain by immunohistochemistry and acquire the concentration of IL-10 in the serum by ELISA. Results show, with the development of IBD, there is a lower expression level of IL-10 in the colon. When rats recover, the expression level of IL-10 increases gradually, and finally reach the normal level. At the same time, the expression level of IL-10 in the cerebral cortex and hypothalamus has a opposite change during the whole process of the IBD. The concentration of IL-10 in the serum begins to lower when IBD happens, on the contrary , it begins to increase accompanied with diease recovery. The experiment shows that IBD is closely related with the activity of neuroendocrine. Immune-nerve-endocrine network may be involved in the process of the occurrence and development of IBD, and it plays a role in the pathogenesis of IBD. Experiment provides a theoretical basis for the study of pathogenesis.
引文
[1]朱长庚,李正莉.细胞因子与免疫一神经一内分泌调节网络[J].解剖学报,1996,27:339-344
[2] Szeleyi J.Cytokines and the sentral nervous system[J].Brain Res Bull,2001,54:329-338
[3] Buer J, Lanoue A , Franzek A,et al.Interleuin 10 secretion and impaired effector function of major histocompatibility complex classⅡ-restricted T cell anergized in vivo[J].J Eur Med,1998,187,177-183
[4]罗克.免疫应答的调节与神经-免疫-内分泌系统的网络调控[J].福建畜牧兽医.2006,28(6):14-17
[5] Moshel,Y.A,Durkin,H.G,Amassian,V.E. Lateralized neocortical control of T lymphocyte export from the thymus:I.Increased export after left cortical stimulation in behaviorally active rats, mediated by sympathetic pathways in the upper spinal cord[J].Neuroimmunol,2005,158:3–13
[6]吉田诚,田中隆一.神经、内分泌、免疫的相互作用[J].脑神经,1996,48(8):703-710
[7] Katayama M,Kobayashis,Kuramoto N,etal.Effects of hy-pothalamic lesions on lympholyte subsets in mice[J].Ann NewYork Acad Sci,1978,496:366-376
[8]戴高中,陈汝兴,顾明昌,等人.大黄虫对大鼠脑出血模型脑组织IL-1βmRNA、iNOSmRNA表达的影响[J].四川中医,2008,26(2):18-20
[9] IL Campbell,MV Hobbs.cerbral expression of multiple cytokine genes in mice lymphocytic choriomeningitis[J].Journal of Immunology,152(2):716-723
[10] Beck Jr,R.D,King,M.A,Ha,G.K, et al.IL-2 deficiency results in altered septal and hippocampal cytoarchitecture:relation to development and neurotrophins[J].Neuroimmunol,2005,160:146–153
[11] Hanisch UK,Seto D,Quirion R.modulation of hippocarnpal acetylcholine release:a potent central action of interleukin-2[J].Neurosci,1993,13:3368-3374
[12] Ching,S,He,L,Lai,W,Quan,N.IL-1 type receptor plays a key role in mediating the recruitment of leukocytes into the central nervous system. Brain Behav[J].Immun,2005,19:127–137
[13]Cambronero,JC,Rivas FJ,Borrell J,Guaza C.interleukin-2 induces corticotropin-releasing hormone release from superfused rat hypothlami:influ nce of glucocorticoids[J]Endocrinology,1992,131:677-683
[14]梁象东,覃雪军,黄春萍,等.糖皮质激素对哮喘患者血清白介素-16和单核细胞趋化蛋白-4浓度的影响[J].内科,2006,1(1):8-10
[15] ArimaM,plittJ,Stellatoc,et al.expression of interleukin-16 by human epithlial cells Inhibition by dexamethasone[J].AMJRespir cell Mol biol,1999,21(6):684-692
[16] Pe.n a AS,Crusius JBA,Oudkerk Pool M,et al.Ge2netics and epidemiology may contribute to understanding the pathogenesis of IBD-a new approach is now indicated[J].Gastroenterol,1993,7:71-74
[17] Darfeuille-Michaud A,Boudeau J,Bulois P,et al.High prevalence of adherent-invasive Escherichia coli associated with ileal mucosa in Crohn’s disease[J].Gastroenterology,2004,127:412–421
[18] Bell AJ,Nicholls RJ,Forbes A,et al.Human lymphocyte stimulation with pouchitis flora is greater than with flora from a healthy pouch but is suppressed by metronidazole[J].Gut,2004,53:1801–1805
[19] Anand AC,Adya CM.Cytokines in inflammatory bowel disease[J].Trop Gast roenterol,1999,20:97-98
[20] Onuma EK,Amenta PS,Ramaswamy K,et al.Autoimmunity in ulcerative colitis (UC):a predominant colonic mucosal B cell response against human tropomyosin isoform 5[J].Clin Exp Immunol,2000,121:466-471
[21] Stevceval L,Pavli P,Husband AJ,et al.The infliammatory infiltrate in the acute stage of the dextran sulphate sodium induced colitis:B cell response differs depending on the percentage of DSS used to induceit[J].BMC Clin Pathol,2001,1:1-3
[22] Rugtveit J,Brandtzaeg P,Halstensen TS,Fausa O,Scott H.increased rectuitment from peripheral blood moncytes[J].Gut,1994,35:669-674
[23] Yao K,Iwashita A,Yao T,Takemura S,Furukawa K,MatsuiT,Aoyagi K,et al.Increased numbers of macrophages in noninflamed gastroduodenal mucosa of patients with Crohn’s disease[J].Dig dis Sci,1996,41:2260-2267
[24] Ashwood P,Harvey R,Verjee T,Wolstencroft R,et al.Functional interactions between mucosal IL-1, IL-ra and TGF-beta 1 in ulcerative colitis[J].Inflamm Res,2004,53:53-59
[25] Mo smann,TR.Cherw insk i,H.Bond,MW.Giedlin,MAand Coffman RL.Two types of murinehelper T cell clone. I. Definit ion according to p rofiles of lymphok ine act ivit ies and secreted p ro teins[J].Immuno,1986,136 (7):2348-2357
[26]邢同京,章廉.Th类细胞极化群体的基础与临床[M].北京:军事医学科学院出版社,2002,108-183
[27] Cosmi L,Annunziato F, Galli M I G et al.CRTH2 is the most reliable marker for the detection of circulating human type 2 Th and type 2 T cyto2 toxic cells in health and disease[J].Eur J Immunol,2000,30 (10):2972-2979
[28] Tsuda H,Michimata T,Sakai M et al.A novel surface molecule of Th2 and Tc2type cells,CRTH2 expression on human peripheral and decidu2 al CD4+ and CD8+ T cells during the early stage of pregnancy[J].ClinExp Immunol,2001,123 (1):105-111
[29] Nagata K,Tanaka K,Ogawa K et al.Selective expression of a novel surface molecule by human Th2 cells in vivo[J].immunol,1999,162(3):1278-1286
[30] Duerr RH,Barmada MM,Zhang IL,et al.High-density genome scan in Crohn disease show confirmed linkage to chromosome 14q11-12[J].Am J Hum Genet,2000,66 (6):1857-1862
[31] Rious JD,Silverberg MS,Daly MJ,et al.Genomewide search in Canadian families with inflammatory bowel disease reveals two novel susceptibility loci[J].Am J Hum Genet,2000,66 (6):1863-18701
[32] Satsangi J.A personal view on progress and prospects[J].Genetics of inflammatory bowel disease.1998,16 (6):370-374
[33] Cuthbert AP,Fisher SA,Mirza MM,et al.The contribution of NOD2 gene mutations to the risk and site of disease in inflammatory bowel disease[J].Gastroenterology,2002,22:867-874
[34] Van Deventer SJ.The future of inflammatory bowel disease therapy[J].Inflamm Bowel Dis,2002,8:301-305
[35] Sanderson JD,Mass MT,Tizard ML,et al.Mycobacterium paratuberculosis DNA in crohn’s disease tissue[J].Gut,1992,33:890– 899
[36] Gionchett P,Rizzello F,renturi A,et al.Oral bacteritherapy as maintenance treatment in patients with chronic pouhitis:A doubleblind,placebo controlled trial[J].Gastroenterol,2000,119:305– 309
[37] Ghosh SS,Gehr TW,Ghosh S,et al1.PPARgamma ligand attenuates PDGF - induded mesangial cell proliferation:role of MAP kinase[J].Kidney Int,2003,64:52– 62
[38] Kim JM.structure of the mouse IL-10 gene and chromosomal localization of the mouse and human genes[J].Immunol,1992,14:3618-3623
[39] Zdanov A.Crystal structure of interlukin-10 reveals the fumctional dimer withan unexpected topological similarity to interferonγ[J].Structure,1995,3(6):591-601
[40] M Parkes,J Satsangi, and D Jewell.Contribution of the IL-2 and IL-10 genes to inflammatory bowel disease (IBD) susceptibility[J].Clin Exp Immunol,1998,113(1):28-32
[41] Tagore A,Gonsalkorale WM,Pravica V,et al.Tissue Antigens,1999,54 :386-390
[42] T Kucharzik,R Stoll,N Lugering,and W Domschke.Circulating antiinflammatory cytokine IL-10 in patients with inflammatory bowel disease (IBD)[J].Clin Exp Immunol,1995,100(3):452-456
[43] Andrew Burich,Robert Hershberg,Kim Waggie,Weiping Zeng,Thea Brabb,Gina Westrich,Joanne L,Viney, Lillian Maggio-Price.Helicobacter-in duced.Inflammatory bowel disease in IL-10 and T cell-deficient mice[J].Am J Physiol Gastrointest Liver Physiol,Sep 2001,281:764-766
[44] N Lugering,T Kucharzik,H Stein,G Winde,A Lugering,A Hasilik,W Domschke,and R Stoll.IL-10 synergizes with IL-4 and IL-13 in inhibiting lysosomal enzyme secretion by human monocytes and lamina propria mononuclear cells from patients with inflammatory bowel[J].Dig Dis Sci,1998,43(4):706-714
[45] MW Leach,NJ Davidson,MM Fort,F Powrie,DM Rennick.The role of IL-10 in inflammatory bowel disease of mice and men[J].Toxicol Pathol, 1999,27(1):123-133
[46] Edward Balish and Thomas Warner.Enterococcus faecalis Induces Inflammatory Bowel Disease in Interleukin-10 Knockout Mice[J].Am. J. Pathol,2002,160:2253
[47] S Schreiber,T Heinig,HG Thiele,and A Raedler.Immunoregulatory role of interleukin 10 in patients with inflammatory bowel disease[J].Gastroenterology,1995,108(5):1434-1444
[48] Fedorak RN,Gangl A,ElsonCO,et al.Recombinant human interleukin-10 in the treatment of patients with mild to moderately active Crohn’sdisease[J].Gastroenteroloty,2000,119(6):1473-1476
[49] Schreiber S,Fedorak RN,Nielsen OH,Wild G,Williams CN,Nikolaus S,Jacyna M,Lashner BA,Gangl A,Rutgeerts P,Isaacs K,van Deventer SJ,Koningsberger JC,Cohard M,LeBeaut A,Hanauer SB.Safety and efficacy of recombinant human interleukin 10 in chronic active Crohn’s disease . Crohs Disease IL-10 Cooperative Study Group[J].Gastroenterology,2000,119:1461-1472
[50]陈英群,董福轮,马贵同.三硝基笨磺酸诱导大鼠溃疡性结肠炎的实验研究[J].同济大学学报,2006,27(6):31-33
[51]刑建峰,侯家玉,邹节明,等人.溃结康胶囊对TNBS诱导大鼠结肠炎治疗作用的研究[J].中药材,2005,28(4):315-318
[52] Butzner JD,Parmar R,Bell CJ,Dalal V.Butyrate enema therapy stimulates mucosal repair in experimental colitis in the rat[J].Gut,1996,38:568-573
[53] Dieleman LA,Palmen MJ,Akol H,Bloemena E,Pena AS,Meuwissen SG,Van Rees EP.Chronic experimental colitis induced by dextran sulphate sodium(DSS) is characterized by Th1 and Th2 cytokines[J].Clin Exp immunol,1998,114:385-391
[54]高庆华,廖圣宝.温脾清肠饮对溃疡性结肠炎患者血清细胞因子IL-6、IL-8、TNF-α的影响[J].中国中医药科技,2008,5(1):12-13
[55] Pooran N,IndaramA,SinfhP,etal.cytokines IL-6、IL -8、TNF-αearly and reliable predictors of severe acute pancreatitis[J].clin Gastroenterol,2003,37:262-266
[56] Koziel MJ.Cvtokines in viral hepatitis[J].semin liver Dis,1999,19:157-169
[57]Rennick DM,fortMM.Am Jphysiol Gastoi-test liver physiol,2000,278(6):829-833
[58] Frank Autschbach,Jutta Braunstein,In Situ Expression of Interleukin-10 in Noninflamed Human Gut and in Inflammatory Bowel Disease[J].American Journal of Pathbology,1998,153:121-130
[59]Meresse B,Rutgeerts P,Malchow H,et al.Low ileal inter-leukin 10 concentrations are predictive of endoscopic recur-rence in patients with Crohn disease[J].GUT,2002,50(1):25
[60] Gasche, S Bakos,IL-10 secretion and sensitivity in normal human intestine and inflammatory bowel disease[J].J Clin Immunol,2000,20(5): 362-370
[61] Cui HH, Chen CL, Wang JD, et al.Effects of p robiotic on intestinalmu2 cosa of patients with ulcerative colitis[J].World J Gastroenterol,2004,10,1521-1525
[62] Niessner M,Volk BA.Altered Th1/Th2 cytokine profiles in the intestinal mucosa of patients with inflammatory bowel disease as assessed by quantitative reversed transcribed polymerase chain reaction (RT-PCR) [J].Clin Exp Immunol,1995,101:428-435
[63] Autschbach F,Braunstein J,Helmke B,et al.In Situ Expression of Interleukin-10 in Noninflamed Human Gut and in Inflammatory Bowel Disease[J].Am J Pathol,1998,153:121-130
[64]Howard M,DgarraA.Biological properties of interleukin-10[J].immunol to day ,1992,13(2):198
[65]李晨.中药肠宁汤对溃疡性结肠炎大鼠IL -10和TNF-α的影响[D].硕士论文,2006
[66]葛相栓,吴正祥,吴强,等人.垂盆草对实验性结肠炎的保护作用及其机制研究[J].中国中西医结合消化杂志,2007,15(6):391-394
[67] Fedorak RN,GanglA,Elson co,et al.Recombinant human interleukin-10 in the treatment of patients with mild to mod-erately active croh’s diease[J].Gastroenterology,2000,119(6):1473
[68] Kronfol Z,Remick DG.cytokines and the brain:implications for clinical psychiatry[J],AmJ psychiatry ,2000,157(5):683-694
[69] Layes,GheusiG,cremonas,et al.Endogenous brain IL-1 mediates lps-induced anorexia and hypothalamic cytokine expression[J].AM Jphysiol,2000,279(1):93-98
[70]Czocchina , Gspiga , SJRabin , Mgrekova , Jrichert , ochenyshev , ccoltonandI moccheei[J].Neurochem Int,1997,30(4):433-439
[71] Kfrei,H Lins,and A Fontana.productin and function of IL-10 in the centaral nervous system[J].Schweiz Arch Neurol psychiaatr,1994,145(3):30-31
[72] Smith,E.M,Cadet,P,Stefano,G.B,Opp,M.R,Hughes Jr,T.K. IL-10 as a mediator in the HPA axis and brain[J].Neuroimmunol,1999,100:140-180
[73] Kennedy,M.K,D.S Torrance,k.s.picha,k.m.mohler.Analysis of cytokine mRNA expression in the central nervous system of mice with exprimental autoimmune encephalomy elities reveals that IL-10 mRNA expression corrcates with recovery[J].immunol,1992,149:2496
[74] Cannella,B,Y.L.Gao,C.Brosnan,c.s.Raine.IL-10 fails to ab-rogate experimental autoimmune encephalomyelitis[J].Neurosci Res,1996,45:735
[75]李真姬.针刺对福而马林疼痛大鼠中枢神经系统c-fos、hsp-70、IL-1的影响[D],硕士论文,2006
[76]李改丽、王百忍.腹腔内注射细菌毒素脂多糖(LPS)或葡萄球菌肠毒素B(SEB)诱发I型IL-1受体在大鼠下丘脑室旁核和视上核表达增强[J].中国组织化学与细胞化学杂志,2002,11(4):362-363
[77] D Agnello,P Villa,P Ghezzi.increased tumor necrosis and interleukin-6 production in the central nervous system of interleukin-10-deficiene mice[J].Brain Res,2000,869(1-2):241-243
[78] AJ Kastin,VAKerstrom,W pan.Interleukin-10 as a CNS therapeutic:the obstacle of the blood-brain/blood-spinal cord barrier[J].Brain Res Mol Brain Res,2003,114(2):168-171