Filipin Ⅲ对肺泡巨噬细胞caveolin-1的影响
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摘要
目的:探索Filipin Ⅲ对肺泡巨噬细胞caveolin-1的影响。方法:采用肺泡灌洗的方法分离提取小鼠原代肺泡巨噬细胞,免疫细胞化学染色鉴定其纯度,Western印迹分析Filipin Ⅲ处理30min后caveolin-1蛋白水平的变化。利用细胞系RAW264.7观察Filipin Ⅲ处理后15min、30min、1h、2h、4h、6h的caveolin-1的动态变化。结果:Filipin Ⅲ处理30min后原代肺泡巨噬细胞及RAW264.7细胞caveolin-1的蛋白水平都有增加。结论:Filipin Ⅲ可以提高肺泡巨噬细胞及RAW264.7细胞caveolin-1的蛋白表达。
Objective to investigate the effect of FilipinⅢ on caveolin-1 expression in alveolar macrophages(AMs).Methods Bronchoalveolar lavage was adopted to extract the primary AMs in mice,which is followed by immunocytochemistry staining to ensure AMs purity.The protein levels of caveolin-1 in AMs treated by FilipinⅢ or DMSO were detected with Western blot.RAW264.7 cells treated by FilipinⅢ were used to observe the possible change of caveolin-1 15 min,30 min,1 h,2 h,4h,6 h later.Results The data showed that caveolin-1 protein levels increased in both AMs and RAW264.7 cells after FilipinⅢ-treatment for 30 min.Conclusion FilipinⅢ can increase the expression of caveolin-1 in AMs and RAW264.7 cells.
Objective to investigate the effect of FilipinⅢ on caveolin-1 expression in alveolar macrophages(AMs).Methods Bronchoalveolar lavage was adopted to extract the primary AMs in mice,which is followed by immunocytochemistry staining to ensure AMs purity.The protein levels of caveolin-1 in AMs treated by FilipinⅢ or DMSO were detected with Western blot.RAW264.7 cells treated by FilipinⅢ were used to observe the possible change of caveolin-1 15 min,30 min,1 h,2 h,4h,6 h later.Results The data showed that caveolin-1 protein levels increased in both AMs and RAW264.7 cells after FilipinⅢ-treatment for 30 min.Conclusion FilipinⅢ can increase the expression of caveolin-1 in AMs and RAW264.7 cells.
引文
[1]Mc Gookey DJ,Fagerberg K,Anderson RG.Filipin-cholesterol complexes form in uncoated vesicle membrane derived from coated vesicles during receptor-mediated endocytosis of low density lipoprotein[J].J Cell Biol,1983.96(5):1273-1278.
[2]Anderson RG.The caveolaemembrane system[J].Annu Rev Biochem,1998,67:199–225.
[3]Fortin DL,Troyer MD,Nakamura K,et al.Lipid rafts mediate the synaptic localization of alpha-synuclein[J].J Neurosci,2004,24(30):6715-6723.
[4]Awasthi-Kalia M,Schnetkam PP,Deans JP.Differential effects of filipin and methyl-beta-cyelodextrin on B cell receptor signaling[J].Biochem Bio Phys Res Commun,2001,287:77-82.
[5]Vetheijen I,Tourlousse D,Vandetheyden PM,et al.Effect of Saponin and filipin on antagonist binding to AT1 receptors in intact cells[J].Biochem Pharmacol,2004,67(8):1601-1606.
[6]罗小林,梁霄,赵慧,等.Filipin对人脐静脉内皮细胞小凹蛋白-1作用的研究[J].石河子大学学报(自然科学版),2012,30(3):337-339.
[7]Mosser DM,Edwards JP.Exploring the full spectrum of macrophage activation[J].Nat Rev Immunol,2008,8(12):958-969.
[8]Wang XM,Kim HP,Song R,et al.Caveolin-1 confers antiinflammatory effects in murine macrophages via the MKK3/p38 MAPK pathway[J].Am J Respir Cell Mol Biol,2006,34(4):434-442.
[9]习臻畅,张坚松.LRG1基因在肝细胞癌中的表达情况研究[J].湖南师范大学学报(医学版),2015,12(2):24-27.
[10]Fujino S,Andoh A,Bamba S,et al.Increased expression of interleukin17 in inflammatory bowel disease[J].Gut,2003,52(1):65-70.
[11]Galan C,Woodard G E,Dionisio N,et al.Lipid rafts modulate the activation but not the maintenance of store-operated Ca(2+)entry[J].Biochim Biophys Acta,2010,1803(9);1083-1093.
[12]Norkin LC,Wolfrom SA,Stuart ES.Association of caveolin with Chlamydia trachomatis inclusions at early and late stages of infection[J].Exp Cell Res,2001,266:229–238.
[13]Yang B,Oo TN,Rizzo V.Lipid rafts mediate H2O2prosurvival effects in cultured endothelial cells[J].FASEB J,2006,20(9):1501-1503.
[14]Kiyanagi T,Iwabuchi K,Shimada K,et al.Involvement of cholesterolenriched microdomains in class A scavenger receptor-mediated responses in human macrophages[J].Atherosclerosis,2011.215(1):60-69.
[2]Anderson RG.The caveolaemembrane system[J].Annu Rev Biochem,1998,67:199–225.
[3]Fortin DL,Troyer MD,Nakamura K,et al.Lipid rafts mediate the synaptic localization of alpha-synuclein[J].J Neurosci,2004,24(30):6715-6723.
[4]Awasthi-Kalia M,Schnetkam PP,Deans JP.Differential effects of filipin and methyl-beta-cyelodextrin on B cell receptor signaling[J].Biochem Bio Phys Res Commun,2001,287:77-82.
[5]Vetheijen I,Tourlousse D,Vandetheyden PM,et al.Effect of Saponin and filipin on antagonist binding to AT1 receptors in intact cells[J].Biochem Pharmacol,2004,67(8):1601-1606.
[6]罗小林,梁霄,赵慧,等.Filipin对人脐静脉内皮细胞小凹蛋白-1作用的研究[J].石河子大学学报(自然科学版),2012,30(3):337-339.
[7]Mosser DM,Edwards JP.Exploring the full spectrum of macrophage activation[J].Nat Rev Immunol,2008,8(12):958-969.
[8]Wang XM,Kim HP,Song R,et al.Caveolin-1 confers antiinflammatory effects in murine macrophages via the MKK3/p38 MAPK pathway[J].Am J Respir Cell Mol Biol,2006,34(4):434-442.
[9]习臻畅,张坚松.LRG1基因在肝细胞癌中的表达情况研究[J].湖南师范大学学报(医学版),2015,12(2):24-27.
[10]Fujino S,Andoh A,Bamba S,et al.Increased expression of interleukin17 in inflammatory bowel disease[J].Gut,2003,52(1):65-70.
[11]Galan C,Woodard G E,Dionisio N,et al.Lipid rafts modulate the activation but not the maintenance of store-operated Ca(2+)entry[J].Biochim Biophys Acta,2010,1803(9);1083-1093.
[12]Norkin LC,Wolfrom SA,Stuart ES.Association of caveolin with Chlamydia trachomatis inclusions at early and late stages of infection[J].Exp Cell Res,2001,266:229–238.
[13]Yang B,Oo TN,Rizzo V.Lipid rafts mediate H2O2prosurvival effects in cultured endothelial cells[J].FASEB J,2006,20(9):1501-1503.
[14]Kiyanagi T,Iwabuchi K,Shimada K,et al.Involvement of cholesterolenriched microdomains in class A scavenger receptor-mediated responses in human macrophages[J].Atherosclerosis,2011.215(1):60-69.