BALB/c小鼠SLIT2和ROBO4基因组织特异性表达与分布的研究
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摘要
SLIT2/RO BO4信号通路在炎症血管反应、白细胞渗出和血管再生等方面发挥重要作用。为深入了解SLIT2/ROBO4信号转导途径在炎症性疾病发生发展过程中的作用机制,本研究采用RT-q PCR和免疫组织化学的方法对SLIT2和ROBO4基因在BALB/c小鼠组织特异性的表达和分布进行了研究。结果表明:SLIT2在肾脏、心脏、肺脏和大脑中表达相对较高,在肾脏表达量最高;ROBO4在心脏、肝脏、肺脏和肾脏中表达相对较高,在心脏中表达量最高。免疫组织化学检测表明,SLIT2和ROBO4在心肌细胞、肝细胞、脾脏的内皮细胞、肺泡上皮细胞、肾小管上皮细胞和大脑皮质神经元中表现出相似的阳性定位。研究结果为深入探讨SLIT2/ROBO4信号通路在炎症性疾病发生发展和调控中的分子作用机制提供了基础数据,也为发展基于该通路的器官特异性炎症治疗策略提供了科学依据。
SLIT2/ROBO4 signal pathway plays an important role in inflammatory vascular responses,and exudation of leukocytes and angiogenesis.To investigate the mechanism of SLIT2/ROBO4 signal transduction pathway in the development of inflammatory diseases,the expression pattern and tissue distribution of SLIT2 and ROBO4 were detected by real-time PCR and immunohistochemical analysis.The results showed that the expression level of SLIT2 was mainly expressed in the kidney,heart,lung and brain and it was the highest in kidney,while the expression level of ROBO4 were relatively high in the heart,liver,lung and kidney and it was the highest in the heart.Immunohistochemical detection showed that SLIT2 and ROBO4 had similar positive location in the cardiac muscle cells,liver cells,endothelial cells of the spleen,alveolar epithelial cells,epithelial cells of the renal tubular and cerebral cortex neurons.The results provide basic data to further explore the molecular mechanisms of SLIT2/ROBO4 signaling pathway in the occurrence,development and regulation of inflammatory diseases and provided a scientific basis for new treatment strategies of organ specific inflammation based on this pathway.
SLIT2/ROBO4 signal pathway plays an important role in inflammatory vascular responses,and exudation of leukocytes and angiogenesis.To investigate the mechanism of SLIT2/ROBO4 signal transduction pathway in the development of inflammatory diseases,the expression pattern and tissue distribution of SLIT2 and ROBO4 were detected by real-time PCR and immunohistochemical analysis.The results showed that the expression level of SLIT2 was mainly expressed in the kidney,heart,lung and brain and it was the highest in kidney,while the expression level of ROBO4 were relatively high in the heart,liver,lung and kidney and it was the highest in the heart.Immunohistochemical detection showed that SLIT2 and ROBO4 had similar positive location in the cardiac muscle cells,liver cells,endothelial cells of the spleen,alveolar epithelial cells,epithelial cells of the renal tubular and cerebral cortex neurons.The results provide basic data to further explore the molecular mechanisms of SLIT2/ROBO4 signaling pathway in the occurrence,development and regulation of inflammatory diseases and provided a scientific basis for new treatment strategies of organ specific inflammation based on this pathway.
引文
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[14]CAI H,XUE Y,LI Z,et al.Roundabout4 suppresses glioma-induced endothelial cell proliferation,migration and tube formation in vitro by inhibiting VEGR2-mediated PI3K/AKT and FAK signaling pathw ays[J].Cellular Physiology and Biochemistry,2015,35(5):1689-1705.
[15]HE H,DI Y,LIANG M,et al.The micro RNA-218 and ROBO1signaling axis correlates with the lymphatic metastasis of pancreatic cancer[J].Oncology Reports,2013,30(2):651-658.
[16]SHI R,YANG Z,LIU W,et al.Knockdown of Slit2 promotes growth and motility in gastric cancer cells via activation of AKT/beta-catenin[J].Oncology Reports,2014,31(2):812-818.
[17]WEBB D R.Animal models of human disease:inflammation[J].Biochemical Pharmacology,2014,87(1):121-130.
[18]BURKE-GAFFNEY A,SVERMOYA T,MUMBY S,et al.Raised plasma Robo4 and cardiac surgery-associated acute kidney injury[J].Plo S One,2014,9(10):e111459.
[19]ZHAO H,ANAND A R,GANJU R K.Slit2-Robo4 pathway modulates lipopolysaccharide-induced endothelial inflammation and its expression is dysregulated during endotoxemia[J].Journal of Immunology,2014,192(1):385-393.
[20]WU J Y,FING L,PARK H T,et al.The neuronal repellent Slit inhibits leukocyte chemotaxis induced by chemotactic factors[J].Nature,2001,410(6831):948-952.
[21]KANELLIS J,GARCIA G E,LI P,et al.Modulation of inflammation by slit protein in vivo in experimental crescentic glomerulonephritis[J].The American Journal of Pathology,2004,165(1):341-352.
[22]TIAN R,LIU Z,ZHANG H,et al.Investigation of the regulation of Roundabout4 by hypoxia-inducible factor-1αin microvascular endothelial cells[J].Investigative Ophthalmology&Visual Science,2015,56(4):2586-2594.
[2]DICKSON B J,GILESTRO G F.Regulation of commissural axon pathfinding by slit and its Robo receptors[J].Annual Review of Cell and Developmental Biology,2006,22(1):651-675.
[3]YPSILANTI A R,ZAGAR Y,CH DOTAL A.Moving away from the midline:new developments for Slit and Robo[J].Development,2010,137(12):1939-1952.
[4]李霖,卿国忠.Slit-Robo信号通路作用的研究进展[J].国际病理科学与临床杂志,2013,33(1):72-77.LI Lin,QING Guo-zhong.Advance in research on the function of Slit-Robo pathway[J].International Journal of Pathology and Clinical Medicine,2013,33(1):72-77.(In Chinese)
[5]HUMINIECKI L,GORN M,SUCHTING S,et al.Magic roundabout is a new member of the roundabout receptor family that is endothelial specific and expressed at sites of active angiogenesis[J].Genomics,2002,79(4):547-552.
[6]YADAV S S,NARAYAN G.Role of ROBO4 signalling in developmental and pathological angiogenesis[J].Bio Med Research International,2014,2014:1-9.
[7]CHATURVEDI S,YUEN D A,BAJWA A,et al.Slit2 prevents neutrophilrecruitmentandrenalischemiar eperfusion injury[J].Journal of the American Society of Nephrology,2013,24(8):1274-1287.
[8]杨明涛,王浩宇,刘涛.Slit-Robo信号通路在心脏血管生成中的作用研究进展[J].中国心血管杂志,2011,16(3):235-237.YANG Ming-tao,WANG Hao-yu,LIU Tao.Advance studies of Slit-Robo signal pathway in cardiac angiogenesis[J].Chinese Journalof Cardiovascular Medicine,2011,16(3):235-237.(In Chinese)
[9]SHORT K R,VELDHUIS KROEZE E J,REPERANT L A,et al.Influenza virus and endothelial cells:a species specific relationship[J].Frontiers in Microbiology,2014,5:653.
[10]JONES C A,LONDON N R,CHEN H,et al.Robo4 stabilizes the vascular network by inhibiting pathologic angiogenesis and endothelial hyperpermeability[J].Nature Medicine,2008,14(4):448-453.
[11]WANG L,ZHANG Z G,GREGG S R,et al.The sonic hedgehog pathway mediates carbamylated erythropoietinenhanced proliferation and differentiation of adult neural progenitor cells[J].The Journal of Biological Chemistry,2007,282(44):32462-32470.
[12]LONDON N R,ZHU W,BOZZA F A,et al.Targeting Robo4-dependent Slit signaling to survive the cytokine storm in sepsis and influenza[J].Science Translational Medicine,2010,2(23):23ra19.
[13]ZHANG X,YU J,KUZONTKOSKI P M,et al.Slit2/Robo4signaling modulates HIV-1 gp120-induced lympha tic hyperpermeability[J].PLo S Pathogens,2012,8(1):e1002461.
[14]CAI H,XUE Y,LI Z,et al.Roundabout4 suppresses glioma-induced endothelial cell proliferation,migration and tube formation in vitro by inhibiting VEGR2-mediated PI3K/AKT and FAK signaling pathw ays[J].Cellular Physiology and Biochemistry,2015,35(5):1689-1705.
[15]HE H,DI Y,LIANG M,et al.The micro RNA-218 and ROBO1signaling axis correlates with the lymphatic metastasis of pancreatic cancer[J].Oncology Reports,2013,30(2):651-658.
[16]SHI R,YANG Z,LIU W,et al.Knockdown of Slit2 promotes growth and motility in gastric cancer cells via activation of AKT/beta-catenin[J].Oncology Reports,2014,31(2):812-818.
[17]WEBB D R.Animal models of human disease:inflammation[J].Biochemical Pharmacology,2014,87(1):121-130.
[18]BURKE-GAFFNEY A,SVERMOYA T,MUMBY S,et al.Raised plasma Robo4 and cardiac surgery-associated acute kidney injury[J].Plo S One,2014,9(10):e111459.
[19]ZHAO H,ANAND A R,GANJU R K.Slit2-Robo4 pathway modulates lipopolysaccharide-induced endothelial inflammation and its expression is dysregulated during endotoxemia[J].Journal of Immunology,2014,192(1):385-393.
[20]WU J Y,FING L,PARK H T,et al.The neuronal repellent Slit inhibits leukocyte chemotaxis induced by chemotactic factors[J].Nature,2001,410(6831):948-952.
[21]KANELLIS J,GARCIA G E,LI P,et al.Modulation of inflammation by slit protein in vivo in experimental crescentic glomerulonephritis[J].The American Journal of Pathology,2004,165(1):341-352.
[22]TIAN R,LIU Z,ZHANG H,et al.Investigation of the regulation of Roundabout4 by hypoxia-inducible factor-1αin microvascular endothelial cells[J].Investigative Ophthalmology&Visual Science,2015,56(4):2586-2594.