摘要
目的探讨干细胞因子(stem cell factor, SCF)和肥大细胞(mast cell, MC)与慢性。肾衰竭大鼠皮肤损害发生发展的关系。
方法46只雄性Wistar大鼠随机分为两组：模型组(n=28)用腺嘌呤150mg·kg-1·d-1灌胃诱发大鼠慢性肾衰竭,对照组(n=18)用等量生理盐水灌胃。观察大鼠一般情况,测量血压。分别于实验第4、8和12周各组随机处死6只大鼠,检测大鼠肾功能,甲苯胺蓝染色观察皮肤组织中MC的浸润,免疫组织化学方法和实时荧光定量PCR检测皮肤组织中SCF蛋白和mRNA的表达量,并分析它们之间的相关性。
结果随着灌胃天数的延长,模型组大鼠一般情况逐渐恶化,血压逐渐升高,贫血逐渐加重,肾功能进行性减退。从第8周开始,模型组大鼠皮肤组织中MC密度和SCF的表达量均明显高于对照组(10.00±1.51vs6.80±0.75,11.78±1.41vs7.17±0.77;0.196±0.040vs0.048±0.013,0.558±0.060vs0.052±0.012;2.64±1.12vsl.08±0.41,6.43±3.53vs1.03±0.30,均P<0.01),且二者均随着灌胃天数的延长逐渐增加。模型组MC密度与SCF蛋白及mRNA的表达量均呈正相关(r分别为0.81、0.65,均P<0.01)。
结论应用腺嘌呤150mg·kg-1·d-1灌胃可以成功构建慢性肾衰竭大鼠模型,且模型大鼠皮肤组织中MC的浸润和SCF的表达均显著增多,提示二者可能是慢性肾衰竭皮肤损害发生发展过程中的一对重要参与者。
Objective To investigate the role of stem cell factor(SCF) and mast cell(MC) in the pathogenesis and progression of dermal lesions caused by chronic renal failure.
Methods46Wistar rats were randomly divided into model group(n=28, adenine lavage at a dose of150mg·kg-1·d-1) and control group(n=18, physiological saline lavage at equal volume).6rats from both groups were sacrificed respectively at week4,8and12. General condition and renal function of the rats were observed. The intensity of MC infiltration was examined by toluidine blue staining. The expression of SCF was detected by immunohistochemistry and real-time fluorescence quantitative PCR.
Results The general condition and renal function of the rats were deteriorating gradually in model group as the lavage going on. A progressive increase of blood pressure and a decrease of hemoglobin also could be found in model group. Compared with control group, the intensity of MC and the expression of SCF were significantly higher in dermal tissue(10.00±1.51vs6.80±0.75,11.78±1.41vs7.17±0.77;0.196±0.040vs0.048±0.013,0.558±0.060vs0.052±0.012;2.64±1.12vs1.08±0.41,6.43±3.53vs1.03±0.30, P<0.01) of model group, and they were increasing as the lavage going on. In the model group, the number of MC infiltration was positively correlated with both the protein expression of SCF(r=0.81, P<0.01) and the level of SCF mRNA(r=0.65, P<0.01).
Conclusions12weeks'adenine lavage at a dose of150mg·kg-1·d-1can develop an ideal rat model of chronic renal failure. The increased SCF and MC may participate in the pathogenesis and progression of dermal lesions caused by chronic renal failure.
引文
[1]王吉耀.内科学.人民卫生出版社.2005.
[2]Pisoni RL, Wikstr6m B, Elder SJ, et al. Pruritus in hemodialysis patients:International results from the Dialysis Outcomes and Practice Patterns Study (DOPPS). Nephrol Dial Transplant 2006, 21(12):3495-3505.
[3]Mistik S, Utas S, Ferahba A, et al. An epidemiology study of patients with uremic pruritus. J Eur Acad Dermatol Venereol,2006,20(6):672-678.
[4]Dyachenko P, Shustak A, Rozenman D. Hemodialysis-related pruritus and associated cutaneous manifestations. Int J Dermatal,2006,45(6):664-667.
[5]Mathur VS, Lindberg J, Germain M, et al. A longitudinal study of uremic pruritus in hemodialysis patients. Clin J Am Soc Nephrol,2010, (5):1410-1419.
[6]Narita I, Alchi B, Omofiok, et al. Etiology and prognostic significance of severe uremic pruritus in chronic hemodialysis patients. Kidney Int,2006,69(9):1626-1632.
[7]Dugas-Breit S, Schopf P, Dugas M, et al. Baseline serum levels of mast cell tryptase are raised in hemodialysis patients and associated with severity of pruritus. J Dtsch Dermatol Ges,2005, 3(5):343-347.
[8]Mettang T, Fritz P, Weber J, et al. Uremic pruritus in patients on hemodialysis or continuous ambulatory peritoneal dialysis (CAPD)-The role of plasma histamine and skin mast cells. Clin Nephrol,1990,34(3):136-41.
[9]El Kossi MM, Haylor JL, Johnson TS, et al. Stem cell factor in a rat Model of serum nephrotoxic nephritis. Nephron Exp Nephrol,2007,108(1):e1-e10.
[10]刘雪梅,马瑞霞,周海燕,等.干细胞因子在狼疮肾炎患者肾组织中的表达及意义.中华风湿病学杂志,2009,13(6)：376-380.
[11]刘雪梅,朱忠华,邓安国,等.肥大细胞与慢性肾小球肾炎肾间质损害的关系.中华肾脏病杂志,2003,19(5)：286-291.
[12]Brezin AL, Brezin JH, Raible DG, et al. Elevated stem cell factor and alpha-protryptase in uremia. Proceedings of the 55th Annual Meeting of the American Academy of Allergy Asthma & Immunology. J Allergy Clin Immunol,1999,103:89.
[13]Kimmel M, Alscher DU, Dunst R, et al. The role of microinflammation in the pathogenesis of uremic pruritus in haemodialysis patients. Nephrol Dial Transplant,2006,21(3):749—755.
[14]Umuechi H, Togashi Y, Honda T, et al. Involvement of central μ-opioid system in the scratching behavior in mice, and the suppression of itch by the activation of μ-opioid system. Eur J Pharmacol,2003,477(1):29—35.
[15]Morton CA, Lafferty M, Hau C, et al. Pruritus and skin hydration during dialysis. Nephrol Dial Transplant,1996,11(10):2031-2036.
[16]Carmichael AJ, Dickinson F, MclHugh MI, et al. Magnesium free dialysis for uremic pruritus. BMJ,1988,297(6663):1584-1585.
[17]Dimkovic N, Djukanovic L, Radmilovic A, et al. Uremic pruritus and skin mast cells. Nephron,1992,61(1):5-9.
[18]Szepietowski JC, Morita A, Tsuji T. Ultraviolet B induces mast cell apoptosis:a hypothetical mechanism of ultraviolet B treatment for uraemic pruritus. Med Hypotheses,2002,58(2): 167-170.
[19]Cohen EP, Russell TJ, Garancis JC. Mast cells and calcium in severe uremic itching. Am J Med Sci,1992,303(6):360-365.
[20]Wershil BK, Furata GT, Lavigne JA, et al. Multiple mechanisms of inhibition of IgE and mast cell dependent cutaneous inflammation in the mouse. J Immunol,1995,154:1391-1398.
[21]郑浩,李卓娅,龚非力,等.跨膜型SCF与分泌型SCF对MC生物学功能影响的研究.中国免疫学杂志,2001,17(8)：404-407.
[22]Jamur MC, Oliver C. Origin, maturation and recruitment of mast cell precursors. Front Biosci, 2011,3:1390-1406.
[23]Laidlaw TM, Steinke JW, Tinana AM, et al. Characterization of a novel human mast cell line that responds to stem cell factor and expresses functional FceRI. J allergy Clin Immunol,2011, 127(3):815-822.
[24]Taylor AM, Galli SJ, Coleman JW. Stem cell factor, the kit ligand, induces direct degranulation of rat peritoneal mast cells in vitro and in vivo:dependence of the in vitro effect on period of culture and comparisons of stem cell factor with other mast cell-activating agents, Immunol,1995,86(3):427-433.
[25]Fujihara M, Azuma H, Ikeda H, et al. Bone marrow stromal cell line promotes the proliferation of mast cell progenitors derived from cord blood CD34+ cell under serum-free conditions with a combination of both cell-cell interaction and soluble factor. Artif Cells Blood Substit Immobil Biotechnol,2011,39(2):51-58.
[26]Torres-Alvarez B, Mesa-Garza LG, Castanedo-Cazares JP, et al. Histochemical and immunohistochemical study in melasma:evidence of damage in the basal membrane. Am J Dermatopathol,2011,33(3):291-295.
[1]Gao C, Kong NR, Chai L. The role of stem cell factor SALL4 in leukemogenesis. Crit Rev Oncoq,2011,16(1-2):117-27.
[2]Martin FH, Suggs SV, Langley KE, et al. Primary structure and functional expression of rat and human stem cell factor DNAs. Cell,1990,63(1):203-211.
[3]郑浩,李卓娅,龚非力,等.跨膜型与分泌型干细胞因子对肥大细胞趋化作用的影响.同济医科大学学报,2001,30(1)：13-15.
[4]Mitkevich VA, Petrushanko IY, Kretova OV, et al. Oncogenic c-kit transcript is a target for binase. Cell Cycle,2010,9(13):2674-2678.
[5]Zhang L, Tang J, Haines CJ, et al. C-kit and its related genes in spermatogonial differentiation. Spermatogenesis,2011,1(3):186-194.
[6]Liu H, Chen X, Focia PJ, et al. Structural basis for stem cell factor-KIT signaling and activation of class III receptor tyrosine kinases. Cell,2007,26(3):891-901.
[7]Duronio V, Scheid MP, Ettinger S. Downstream signaling events regulated by phosphatidylinositol 3-kinase activity. Cell Signal,1998,10(4):233-239.
[8]Maddens S, Charruyer A, Plo I, et al. Kit signaling inhibits the sphingomyelin-ceramide pathway through PLC gamma 1:implication in stem cell factor radio protective effect. Blood, 2002,100(4):1294-1301.
[9]Kalaitzidis D, Neel BG. Flow-cytometric phosphoprotein analysis reveals agonist and temporal differences in responses of murine hematopoietic stem/progenitor cells. PLoS One,2008,3(11): e3776.
[10]Hong L, Munugalavadla V, Kapur R, et al. C-kit mediated overlapping and unique functional and biochemical outcomes via diverse signaling pathways. Mol Cell Biol,2004,24(3):1401-1410.
[11]Roskoskj R. Signaling by Kit protein-tyrosine kinase--the stem cell factor receptor. Biochem Biophys Res Commun,2005,337(1):1-13.
[12]Brannan CI, Lyman SD, williams DE, et al. Steel-Dickie mutation encodes a c-kit ligand lacking transmembrane and cytoplasmic domains. Proc Natl Acad Sci USA,1991,88(11): 4671-4674.
[13]Matsuzawa S, Sakashita K, Kinoshita T, et al. IL-9 enhances the growth of human mast cell progenitors under stimulation with stem cell factor. Immunol,2003,170(7):3461-3467.
[14]Tsai M, Tskeishi T, Thompson H, el al. Induction of mast cell proliferation, maturation, and heparin synthesis by the rat c-kit ligand-stem cell factor. Proc Natl Acad Sci USA,1991,15: 88(14):6382-6386.
[15]Moller C, Alfredsson J, Engstrom M, et al. Stem cell factor promotes mast cell survival via inactivation of F OXO3a-mediated transcriptional induction and MEK-regulated phosphorylation of the proapoptotic protein Bim. Blood,2005,106(4):1330-1336.
[16]Reber L, Da Silva CA, Frossard N. Stem cell factor and its receptor c-Kit as targets for inflammatory diseases. Eur J Pharmacol,2006,533(1-3):327-340.
[17]Takeuchi K, Koike K, Kamijo T, et al. STI571 inhibits growth and adhesion of human mast cells in culture. J Leukoc Biol,2003,74(6):1026-1034.
[18]Huang B, Lei Z, Zhang GM, et al. SCF-mediated mast cell infiltration and activation exacerbate the inflammation and immunosuppression in tumor microenvironment. Blood,2008, 112(4):1269-1279.
[19]Nagata H, Worobec AS, Oh CK, et al. Identification of a point mutation in the catalytic domain of the protooncogene c-kit in peripheral blood mononuclear cells of patients who have mastocytosis with an associated hematologic disorder. Proc Nail Acad Sci USA,1995,92(23): 10560-10564.
[20]Longley BJ, Tyrrell L, Lu SZ, et al. Somatic c-KIT activating mutation in urticaria pigmentosa and aggressive mastocytosis:establishment of clonality in a human mast cell neoplasm. Nat Genet,1996,12(3):312-314.
[21]Pignon JM, Giraudier S, Duquesnoy P, et al. A new c-kit mutation in a case of aggressive mast cell disease. J Haematol,1997,96(2):374-376.
[22]Akin C, Jaffe ES, Raffled M, et al. An immunohistochemical study of the bone marrow lesion of systemic mastocytosis:expression of stem cell factor by lesional mast cells. Am J Clin Pathol, 2002,118(2):242-247.
[23]张利中,呼和,徐敏丽,等.干细胞因子SCF在肥大细胞增生病患者皮肤组织中蛋白水平的表达.中华微生物学和免疫学杂志,2006,26(12)：1048.
[24]Wang J, Feng J, Wang W, et al. Role of stem cell factor and its receptor in the pathogenesis of pediatric aplastic anemia. J Huazhong Univ Sci Technolog Med Sci,2005,25(1):29-31.
[25]张云娟,王军,王伟,等.干细胞因子及其受体与儿童再生障碍性贫血关系的研究.中华血液学杂志,2008,29(7)：484-486.
[26]Kojima S, Matsuyama T, Kodem Y. Plasma levels and production of soluble stem cell factor by marrow stromal cells in patients with aplastic anemia. Br J Haematol,1997,99:440446.
[27]Al-Muhsen SZ, Shablovsky G, Olivenstein R, et al. The expression of stem cell factor and c-kit receptor in human asthmatic airways. Clin Exp Allergy,2004,34(6):911-916.
[28]Da Silva CA, Blay F, Israel-Biet D, et al. Effect of glucocorticoids on stem cell factor (SCF) expression in human asthmatic bronchi. Clin Exp Allergy,2006,36(3):317-324.
[29]Makowska JS, Cieslak M, Kowalski ML. Stem cell factor and its soluble receptor (c-kit) in serum of asthmatic patients-correlation with disease severity. BMC Pulm Med,2009,9:27.
[30]Metcalf D, Mifsud S, Di RagoL. Stem cell factor can stimulate the formation of eosinophils by two types of marine eosinophil progenitor cells. Stem Cells,2002,20(5):460-469.
[31]Yamamoto T, Katayama I, Nishioka K. Possible contribution of stem cell factor in psoriasis vulgaris. J Dermatol Sci,2000,24(3):171-176.
[32]刘允辉阎春林廖康煌.银屑病患者皮损微血管密度和肥大细胞密度及光化学疗法对其影响.中国现代医学杂志,2006,16(19)：2897-2899.
[33]Das RP, Jain AK, Ramesh V. Current concepts in the pathogenesis of psoriasis. Indian J Dermatol,2009,54(1):7-12.
[34]Osuga Y, Koga K, Tsutsumi O, et al. Stem cell factor (SCF) concentrations in peritoneal fluid of women with or without endometriosis. Am J Reprod Immunol,2000,44(4):231-235.
[35]Anaf V, Chapron C, El Nakadi I, et al. Pain, mast cells, and nerves in peritoneal, ovarian, and deep infiltrating endometriosis. Fertil Steril,2006,86(5):1336-1343.
[36]Konno R, Yamada-Okabe H, Fujiwara H, et al. Role of immunoreactions and mast cells in pathogenesis of human endometriosis-morphologic study and gene expression analysis. Hum Cell, 2003,16(3):141-149.
[37]Yamada M, Ueda M, Naruko T, et al. Mast cell chymase expression and mast cell phenotypes in human rejected kidneys. Kidney Int,2001,59(4):1374-1381.
[38]Berlin AA, Hogaboam CM, Lukacs NW. Inhibition of SCF attenuates peribronchial remodeling in chronic cockroach allergen-induced asthma. Lab Ivest,2006,86(6):7-565.
[39]Xu X, Pappo O, Garbuzenko E, et al. Mast cell dynamics and involvement in the development of peritoneal adhesions in the rat. Life Sci,2002,70(8):951-967.
[40]刘雪梅,朱忠华,邓安国,等.肥大细胞与慢性肾小球肾炎肾间质损害的关系.中华肾脏病杂志,2003,19(5)：286-291.
[41]刘雪梅,马瑞霞,朱月华,等.肥大细胞、巨噬细胞及转化生长因子在IgA肾病患者肾组织中的浸润及意义.中国组织工程研究与临床康复,2007,11(8)：1485-1488.
[42]李瑛刘伏友周乐天,等.肥大细胞参与蛋白负荷肾病大鼠肾间质的纤维化.中华肾脏病杂志,2008,24(7)：498-503.
[43]Gaca MD, Pickering JA, Arthur MJ. Human and rat hepatic satellite cells produce stem cell factor:a possible mechanism for mast cell recruitment in liver fibrosis. J Hepatol,1999 30(5): 850-858.
[44]Yamamoto T, Hartmann K, Eckes B, et al. Role of stem cell factor and monocyte chemoattractant protein-1 in the interaction between fibroblasts and mast cells in fibrosis. J Dermatol Sci,2001,26(2):106-111.