PDF(1313 KB)
孟鲁司特和卡介苗多糖核酸对哮喘大鼠血单个核细胞中STAT5b mRNA和IL-4 mRNA表达的影响
金耀建, 宋志燕, 胡野, 钱旭波, 王晓杨, 何旭瑛
中国当代儿科杂志 ›› 2009, Vol. 11 ›› Issue (02) : 133-137.
PDF(1313 KB)
PDF(1313 KB)
孟鲁司特和卡介苗多糖核酸对哮喘大鼠血单个核细胞中STAT5b mRNA和IL-4 mRNA表达的影响
Effects of montelukast and BCG-PSN on the expression of STAT5b mRNA and IL-4 mRNA in blood mononuclearcells of rats with asthma
目的：研究孟鲁司特和卡介苗多糖核酸对哮喘大鼠血单个核细胞中信号转导子和转录激活子5b(STAT5b) mRNA和白细胞介素4（IL-4）mRNA的转录表达的影响。方法：52只体重为140~200 g清洁级Sprague-Dawley雄性大鼠，随机分为4组：哮喘组、孟鲁司特组、卡介苗多糖核酸组和正常对照组。用卵白蛋白制备大鼠哮喘模型。计数血中嗜酸性粒细胞(EOS)；应用双抗体夹心酶联免疫吸附试验测定血浆中IL-4和γ-干扰素（IFN-γ）浓度；采用SYBR GREEN I荧光实时定量PCR法测定STAT5b mRNA和IL-4 mRNA的相对表达量。结果：①哮喘组血单个核细胞中STAT5b mRNA和IL-4 mRNA的表达量高于其他各组（P<0.01）；在孟鲁司特组和卡介苗多糖核酸组中两者的表达量相近（P>0.05）。哮喘组除了IFN-γ水平低外，其余无论是STAT5b mRNA，IL-4 mRNA，EOS绝对值等都是4组中最高的（P<0.01）；除哮喘组外，其余组的各项指标相近（P>0.05）。②STAT5b mRNA表达量与IL-4 mRNA表达量、IL-4浓度、EOS绝对值呈正相关(P<0.01)，与IFN-γ浓度呈负相关（P<0.01）。结论：哮喘大鼠血单个核细胞中STAT5b mRNA和IL-4 mRNA的表达增强且呈正相关；孟鲁司特和卡介苗多糖核酸可以下调STAT5b mRNA和IL-4 mRNA的表达，可能为其抑制哮喘气道炎症形成的重要机制之一。［中国当代儿科杂志，2009，11（2）：133-137］
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］
哮喘 / 孟鲁司特 / 卡介苗多糖核酸 / STAT5b mRNA / IL-4 mRNA / 大鼠
Asthma / Montelukast / BCG-PSN / STAT5b mRNA / IL-4 mRNA / Rats
［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.
［5］杨玉成，洪苏玲，黄江菊，王祎琴，孙荣，钱迪，等．卡介苗多糖核酸对变应性鼻炎病人Th1/Th2失衡状态的调节［J］．第三军医大学学报，2005，27（12）：1279-1281．
［6］冼乐武，梁宗安，刘春涛．支气管哮喘大鼠单个核细胞Th1/Th2的变化及孟鲁司特的干预作用［J］．中华结核和呼吸杂志，2003，26（5）：312-313．
［7］叶乐平，谢丽微，李昌崇．布地奈德对哮喘大鼠信号转导子和转录激活子6的表达［J］．中华微生物学和免疫学杂志，2005，25（7）：588-593．
［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.
［11］李国平，刘志刚，冉丕鑫，丘劲，钟南山．卵白蛋白诱导哮喘小鼠脾细胞增殖中STAT5的变化［J］．细胞与分子免疫学杂志，2004，20（5）：611-614．
［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.
［24］王曼芝，何庆南，袁红霞，刘晓亮.IL-4， IL-5及IgE在儿童哮喘变异性哮喘中的价值［J］.中国当代儿科杂志，2006， 8（5）：382-384.
［25］李光，赵育莹，曲书强，逮晓辉，刘晨，唐建民，等.支气管哮喘模型大鼠IL-4，IL-10，IFN-γ的检测［J］.中国当代儿科杂志，2004， 6（6）：474-476.
