Pyk2、CXCR_1在COPD大鼠气道炎症反应中的表达及意义
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摘要
目的:探讨脯氨酸蛋白酪氨酸激酶(Pyk2)、化学趋化因子受体1 (CXCR_1)在慢性阻塞性肺疾病(COPD)大鼠气道炎症反应中的表达及意义.方法:SD大鼠随机分为正常组和模型组,模型组以气管内注入内毒素脂多糖(LPS)联合烟熏构建COPD大鼠模型,利用光学显微镜观察模型组与正常组大鼠肺组织病理结构差别;ARMS-实时荧光定量PCR检测Pyk2、CXCR_1m RNA表达水平; Western blot检测肺组织中Pyk2、CXCR_1蛋白水平.结果:(1)与模型组比较,正常组大鼠肺组织炎症细胞浸润较少,肺泡壁结构正常.模型组肺组织部分纤维化,大量炎症细胞尤以巨噬细胞为主,充斥于肺泡腔.(2)正常组Pyk2、CXCR_1m RNA表达水平较模型组低,其表达丰度分别为模型组的0. 099、0. 614.(3)正常组Pyk2、CXCR_1蛋白表达较模型组减少.结论:Pyk2、CXCR_1在COPD大鼠中呈高表达状态,提示两者在COPD大鼠气道炎症反应机制中扮演重要角色.
Objective: To investigate the expression and significance of proline-rich tyrosine kinase 2( Pyk2) and CXC chemokine receptor 1( CXCR_1) in airway inflammation of COPD rats. Methods: SD rats were randomly divided into a normal group and a model group. In model group,the rats were injected with endotoxin lipopolysaccharide( LPS) into trachea and exposed to cigarette smoke to establish COPD model. The difference in the structure of the lung tissue between the model group and the normal group was observed under optical microscope. The expression level of Pyk2 and CXCR_1 m RNA was detected by real-time PCR. The level of Pyk2 and CXCR_1 in lung tissue were detected by Western blot assay. Results: Compared with model group,less inflammatory cells infiltrated in lung tissue of normal group rats,and the alveolar wall structure was normal. In model group,the lung tissue was partially fibrotic,and a large number of inflammatory cells,mainly macrophages,filled the alveolar cavity. Theexpression level of Pyk2 and CXCR_1 m RNA in normal group were lower than that in model group,and the expression abundance was 0. 099 and 0. 614 of model group,respectively. The expression of Pyk2 and CXCR_1protein in normal group was lower than that in model group. Conclusion: Pyk2 and CXCR_1 are highly expressed in COPD rats,indicating that they play an important role in the airway inflammation response mechanism in COPD rats.
Objective: To investigate the expression and significance of proline-rich tyrosine kinase 2( Pyk2) and CXC chemokine receptor 1( CXCR_1) in airway inflammation of COPD rats. Methods: SD rats were randomly divided into a normal group and a model group. In model group,the rats were injected with endotoxin lipopolysaccharide( LPS) into trachea and exposed to cigarette smoke to establish COPD model. The difference in the structure of the lung tissue between the model group and the normal group was observed under optical microscope. The expression level of Pyk2 and CXCR_1 m RNA was detected by real-time PCR. The level of Pyk2 and CXCR_1 in lung tissue were detected by Western blot assay. Results: Compared with model group,less inflammatory cells infiltrated in lung tissue of normal group rats,and the alveolar wall structure was normal. In model group,the lung tissue was partially fibrotic,and a large number of inflammatory cells,mainly macrophages,filled the alveolar cavity. Theexpression level of Pyk2 and CXCR_1 m RNA in normal group were lower than that in model group,and the expression abundance was 0. 099 and 0. 614 of model group,respectively. The expression of Pyk2 and CXCR_1protein in normal group was lower than that in model group. Conclusion: Pyk2 and CXCR_1 are highly expressed in COPD rats,indicating that they play an important role in the airway inflammation response mechanism in COPD rats.
引文
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[2]BERENSON C S,KRUZEL R L,EBERHARDT E,et al.Phagocytic dysfunction of human alveolar macrophages and severity of chronic obstructive pulmonary disease[J].The Journal of Infectious Diseases,2013,12(12):2036-2045.
[3]GREGEORY A D,KLIMENT C R,METZ H E,et al.Neutrophil elastase promotes myofibroblast differentiation in lung fibrosis[J].Journal of Leukocyte Biology:An Official Publication of the Reticuloendothelial Society,2015,2(2):143-152.
[4]SIEG D J,ILIC D,JONES K C,et al.Pyk2 and Srcfamily protein-tyrosine kinases compensate for the loss of FAK in fibronectin-stimulated signaling events but Pyk2 does not fully function to enhance FAK-cell migration[J].EMBO Journal,1998,20(20):5933-5947.
[5]DUAN Y,JONATHAN L,ANGELO Y M,et al.Inhibition of Pyk2 blocks lung inflammation and injury in a mouse model of acute lung injury[J].Respiratory Research,2012,13(1):4.
[6]OWEN K A,PIXLEY F J,THOMAS K S.Regulation of lamellipodial persistence,adhesion turnover,and motility in macrophages by focal adhesion kinase[J].J Cell Biol,2007,179(6):1275-1287.
[7]SHINKAI M,GREGORY H F,BRUCE K R.Macrolide antibiotics modulate ERK phosphorylation and IL-8 and GM-CSF production by human bronchial epithelial cells[J].Am J Physiol Lung Cell Mol Physiol,2006,290(1):75-85.
[8]YUO A,ITAGA S,KASAHARA T,et al.Stimulation and priming of human neutrophils by interleukin-8:cooperation with tumor necrosis factor and colonystimulating factors[J].BLOOD,1991,78(10):2708-2714.
[9]WOZNIAK A,BETTS W H,MURPHY G A,et al.Interleukin-8 primes human neutrophils for enhanced superoxide anion production[J].Immunology,1993,79(4):608-615.
[10]METZNER B,BARBISCH M,PARLOW F,et al.Interleukin-8 and GRO alpha prime human neutrophils for superoxide anion production and induce upregulation of N-formyl peptide receptors[J].Journal of Investigative Dermatology,1995,104(5):789-791.
[11]JONES S A,WOLF M,QIN S,et al.Different functions for the interleukin 8 receptors(IL-8R)of human neutrophil leukocytes:NADPH oxidase and phospholipase D are activated through IL-8R1 but not IL-8R2[J].Proceedings of The National Academy of Sciences of The United States of America.1996,93(13):6682-6686.
[12]MCCOLL S R,KREIS C,DIPERSIO J F,et al.Involvement of guanine nucleotide binding proteins in neutrophil activation and priming by GM-CSF[J].BLOOD,1989,73(2):588-591.
[13]SOZZANI S,AGWU D E,ELLENBURG M D,et al.Activation of phospholipase D by interleukin-8 in human neutrophils[J].Blood,1994,84(13):3895-3901.