Abstract OBJECTIVE: To study the expression of signal transducer and activator of transcription 5b (STAT5b) mRNA and interleukin-4 (IL-4) mRNA in blood mononuclearcells in a rat model of asthma and the effect of montelukast (MK) and BCG-polysaccharide and nucleic acid injection (BCG-PSN) on STAT5b mRNA and IL-4 mRNA expression. METHODS: Fifty-two male Sprague-Dawley rats (weight:140-200 g) were randomly divided into four groups: asthma, MK-treated and BCG-PSN-treated and control groups. Rat model of asthma was prepared by ovalbumin (OVA) sensitization. The rats were sacrificed 24 hrs after the last sensitization. Blood eosinophils (EOS) were counted. Plasma contens of IL-4 and interferon-γ (IFN-γ) were measured using ELISA. Expression of STAT5b mRNA and IL-4 mRNA in blood mononuclearcells was detected with SYBR GREEN I fluorescent quantitation PCR method. RESULTS: Blood contents of STAT5b mRNA and IL-4 mRNA in the untreated asthma group were significantly higher than those in the other three groups (P<0.01). Blood EOS count and plasma IL-4 contents in the untreated asthma group significantly increased, while plasma IFN-γ contents significantly decreased compared with the other three groups (P<0.01). There were no significant differences in the parameters measured among the MK-treated, the BCG-PSN-treated and the control groups. STAT5b mRNA expression was positively correlated to IL-4 mRNA expression, IL-4 content and EOS count (r=0.730, 0.650, 0.664, respectively; P<0.01), but negatively correlated to IFN-γ content (r=-0.798; P<0.01). CONCLUSIONS: STAT5b mRNA and IL-4 mRNA were strongly expressed in blood mononuclearcells in rats with asthma, and there was a positive correlation between them. MK and BCG-PSN had inhibitory effects on the expression of STAT5b mRNA and IL-4 mRNA, which might be contributed to suppression of airway inflammation in asthma.［Chin J Contemp Pediatr, 2009, 11 (2):133-137］
JIN Yao-Jian,SONG Zhi-Yan,HU Yu et al. Effects of montelukast and BCG-PSN on the expression of STAT5b mRNA and IL-4 mRNA in blood mononuclearcells of rats with asthma[J]. 中国当代儿科杂志, 2009, 11(02): 133-137.
JIN Yao-Jian,SONG Zhi-Yan,HU Yu et al. Effects of montelukast and BCG-PSN on the expression of STAT5b mRNA and IL-4 mRNA in blood mononuclearcells of rats with asthma[J]. CJCP, 2009, 11(02): 133-137.
［1］Buitenhuis M, Baltus B, Lammers JW, Coffer PJ, Koenderman L. Singal transducer and activator of transcription 5a (STAT5a) is required for eosinophil differentiation of human cord blood-derived CD34+cells［J］. Blood, 2003, 101(1):134-142.
［2］Soldaini E, John S, Moro S, Bollenbacher J, Schindler U, Leonard WJ. DNA binding site selection of dimeric and tetrameric Stat5 proteins reveals a large repertoire of divergent tetrameric Stat5a binding sites［J］. Mol Cell Biol, 2000, 20(1):389-401.
［3］Sergejeva S, Tomaki M, Pullerits T, Zhao LL, Johnson M, L-tvall J. Intransal fluticasone propionate inhibits allergen induce bone marrow eosinophilia in mice［J］. Pulm Pharmacol Ther, 2002, 15(2):129-134.
［4］Takatori H, Nakajima H, Hirose K, Kagami S, Tamachi T, Suto A, et al. Indispensable role of Stat5a in Stat6-independent Th2 cell differentiation and allergic airway inflammation［J］. J Immunol, 2005, 174(6):3734-3740.
［8］Palmans E, Vanacker NJ, Pauwels RA, Kips JC. Effect of age on allergen-induced structural airway changes in brown Norway rats［J］．Am J Respir Crit Care Med, 2002, 165(9):1280-1284．
［9］Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta DeltaC(T)) method［J］. Methods, 2001, 25(4):402-408.
［10］Buitenhuis M, Baltus B, Lammers JW, Coffer PJ, Koenderman L. Singal transducer and activator of transcription 5a (STAT5a) is required for eosinophil differentiation of human cord blood-derived CD34+cells［J］. Blood, 2003, 101(1):134-142.
［12］Zamorano J, Wang HY, Wang R, Shi Y, Longmore GD, Keegan AD. Regulation of cell growth by IL-2: role of STAT5 in protection from apoptosis but not in cell cycle progression［J］. J Immunol, 1998, 160(7):3502-3512.
［13］Zhu Y, Chen L, Huang Z, Alkan S, Bunting KD, Wen R, et al. Cutting edge: IL-5 primes Th2 cytokine-producing capacity in eosinophils through a STAT5dependent mechanism［J］. J Immunol, 2004, 173(5):2918-2922.
［14］Takatori H, Nakajima H, Hirose K, Kagami S, Tamachi T, Suto A, et al. Indispensable role of Stat5a in stat6-independent Th2 cell differentiation and allergic airway inflammation［J］. J Immunol, 2005, 174(6):3734-3740.
［15］Hansen G, Berry G, Dekruyff RH, Umetsu DT. Allergen-specific Th1 cells fail to counterbalance Th2 cell-induced airway hyperreactivity but cause severe airway inflammation［J］. J Clin Invest, 1999, 103(2):175-183.
［16］Lingnau K, Hoehn P, Kerdine S, Koelsch S, Neudoerfl C, Palm N, et al. IL-4 in combination with TGF-beta favors an alternative pathway of Th1 development independent of IL-12［J］. J Immunol, 1998, 161(9): 4709-4718.
［17］Stelmach I, Jerzynska J, Kuna P. A randomized,double-blind trial of the effect of treatment with montelukast on bronchial hyperresponsiveness and serum eosinophilic cationic protein(ECP), soluble interleukin 2 receptor(sIL-2R), IL-4, and soluble intercellular adhesion molecule-1(sICAM-1) in children with asthma［J］. J Allergy Clin Immunol, 2002, 109(2):257-263.
［18］Braccioni F, Dorman SC, O′byrne PM, Inman MD, Denburg JA, Parameswaran K, et al. The effect of cysteinyl leukotrienes on growth of eosinophil progenitors from peripheral blood and bone marrow of atopic subjects［J］. J Allergy Clin Immunol, 2002, 110(1):96-101.
［19］Homer AA, Van Uden JH, Zubeldia JM, Broide D, Raz E. DNA-based immunotherapeutics for the treatment of allergic disease［J］. Immunol Rev, 2001, 179: 102-118.
［20］Ambrosio R, Fimiani G, Monfregola J, Sanzari E, De Felice N, Salerno MC, et al. The structure of human STAT5A and B gene sreveals two regions of nearly identical sequence and an alternative tissue specific STAT5B promoter［J］. Gene, 2002, 285(12):311-318.
［21］Crispi S, Sanzari E, Monfregola J, De Felice N, Fimiani G, Ambrosio R, et al. Characterization of the human STAT5A and STAT5B promoters: evidence of a positive and negative mechanism of transcriptional regulation［J］. FEBS Lett, 2004, 562(1-3):27-34.
［22］Uddin S, Lekmine F, Sassano A, Rui H, Fish EN, Platanias LC. Role of Stat5 in type I interferon-signaling and transcriptional regulation［J］. Biochem Biophys Res Commun, 2003, 308(2):325-330.
［23］Yamashita H, Nevalainen MT, Xu J, LeBaron MJ, Wagner KV, Erwin RA, et al. Role of serine phosphorylation of Stat5a in prolactin-stimulated bata-casein gene expression［J］. Mol Cell Endocrinol, 2001, 183(1-2):151-163.
