Tie2基因对纳米金颗粒进入牙周组织的影响
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摘要
目的研究Tie2基因表达对不同粒径纳米金颗粒(gold nanoparticles,GNPs)进入牙周组织及停留情况的影响。方法采用实时定量PCR和Western印迹法检测Tie2基因和蛋白在正常和转基因小鼠牙周组织中的表达;冰冻切片检测GFP在牙周组织中相对高表达的部位;小鼠尾静脉注射1 g/kg剂量20、40、60、80 nm粒径GNPs,应用ICP-AES技术检测牙周组织Au含量。结果转基因小鼠比正常小鼠Tie2基因的表达高约2. 5倍。Tie2蛋白在转基因小鼠牙周组织中显著高表达,差异具有统计学意义(P<0. 05)。GFP荧光提示Tie2在转基因小鼠牙周组织血管周围表达相对较高。转基因小鼠和正常小鼠牙周组织中GNPs的累积量随粒径增大而减少,正常小鼠牙周组织中GNPs的累积量明显高于转基因小鼠,差异具有统计学意义(P<0. 05)。结论 Tie2基因高表达不利于GNPs扩散。
Objective To investigate the effect of Tie2 gene expression on diffusion of gold nanoparticles( GNPs) into periodontal tissues in mice. Methods The expression of Tie2 mRNA and Tie2 protein in periodontal tissues of normal and transgenic BALB/c mice was detected by real-time RT-PCR and Western Blot. The expression of GFP in periodontal tissue of transgenic Tie2-GFP mice was detected by fluoromicroscopy with frozen section. Mice were injected with GNPs( 1 g/kg) with diameters of 20,40,60 and 80 nm,respectively; and the content of gold( Au) in periodontal tissue was detected by ICP-AES. Results The mRNA expression in periodontal tissue of Tie2 transgenicmice was 2. 5 times higher than that in normal mice; and the expression of Tie2 protein was also higher in the periodontal tissues of transgenic mice( P<0. 05). GFP fluorescence indicated that Tie2 protein was higher expressed around the vessels in periodontal tissue of transgenic mice. The contents of GNPs in the periodontal tissue of transgenic mice and normal mice were decreased with the increase of particle size. The amount of GNPs in periodontal tissue of normal mice was significantly higher than that of transgenic mice( P<0. 05). Conclusion High expression of Tie2 gene is associated with the decreased diffusion of gold nanoparticles into periodontal tissue.
Objective To investigate the effect of Tie2 gene expression on diffusion of gold nanoparticles( GNPs) into periodontal tissues in mice. Methods The expression of Tie2 mRNA and Tie2 protein in periodontal tissues of normal and transgenic BALB/c mice was detected by real-time RT-PCR and Western Blot. The expression of GFP in periodontal tissue of transgenic Tie2-GFP mice was detected by fluoromicroscopy with frozen section. Mice were injected with GNPs( 1 g/kg) with diameters of 20,40,60 and 80 nm,respectively; and the content of gold( Au) in periodontal tissue was detected by ICP-AES. Results The mRNA expression in periodontal tissue of Tie2 transgenicmice was 2. 5 times higher than that in normal mice; and the expression of Tie2 protein was also higher in the periodontal tissues of transgenic mice( P<0. 05). GFP fluorescence indicated that Tie2 protein was higher expressed around the vessels in periodontal tissue of transgenic mice. The contents of GNPs in the periodontal tissue of transgenic mice and normal mice were decreased with the increase of particle size. The amount of GNPs in periodontal tissue of normal mice was significantly higher than that of transgenic mice( P<0. 05). Conclusion High expression of Tie2 gene is associated with the decreased diffusion of gold nanoparticles into periodontal tissue.
引文
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[10]BARTON W A,DALTON A C,SEEGAR T C M,et al.Tie2 and Eph receptor tyrosine kinase activation and signaling[J].Cold Spring Harb Perspect Biol,2014,6(3):a009142.
[11]MOSS A.The angiopoietin:Tie 2 interaction:a potential target for future therapies in human vascular disease[J].Cytokine Growth Factor Rev,2013,24(6):579-592.
[12]LELIGDOWICZ A,RICHARD-GREENBLATT M,WRIGHT J,et al.Endothelial activation:the Ang/Tie axis in sepsis[J].Front Immunol,2018,9:838.
[13]LIMS S,KOOKSH,LEEJ C.COMP-Ang1 enhances DNA synthesis and cell cycle progression in human periodontal ligament cells via Tie2-mediated phosphorylation of PI3K/Akt and MAPKs[J].Mol Cell Biochem,2016,416(1-2):157-168.
[14]CHITHRANI D B.Intracellular uptake,transport,and processing of gold nanostructures[J].Mol Membr Biol,2010,27(7):299-311.
[15]ALKILANY A M,NAGARIA P K,HEXEL C R,et al.Cellular uptake and cytotoxicity of gold nanorods:molecular origin of cytotoxicity and surface effects[J].Small,2009,5(6):701-708.
[2]LI C,LI Z Q,WANG Y,et al.Gold nanoparticles promote proliferation of human periodontal ligament stem cells and have limited effects on cells differentiation[J].J Nanomaterial,2016,2016:1-10.
[3]倪璞,刘宏伟.骨髓间充质干细胞培养液对牙周组织再生的影响[J].同济大学学报(医学版),2018,39(1):41-46
[4]SAHARINENP,EKLUNDL,ALITALO K.Therapeutic targeting of the angiopoietin-TIE pathway[J].Nat Rev Drug Discov,2017,16(9):635-661.
[5]PARTANEN J,ARMSTRONG E,MAKELAP,et al.A novel endothelial cell surface receptor tyrosine kinase with extracellular epidermal growth factor homology domains.[J].Mol Cell Biol,1992,12(4):1698-1707.
[6]JELTSCH M,LEPPANEN V M,SAHARINEN P,et al.Receptor tyrosine kinase-mediated angiogenesis[J].Cold Spring Harb Perspect Biol,2013,5(9):a009183.
[7]KITAJIMA D,KASAMATSU A,NAKASHIMA D,et al.Evidence for critical role of Tie2/Ang1 interaction in metastatic oral cancer[J].Oncol Lett,2018,15(5):7237-7242.
[8]GOMEZ PERDIGUERO E,KLAPPROTH K,SCHULZ C,et al.Tissue-resident macrophages originate from yolk-sac-derived erythro-myeloid progenitors[J].Nature,2015,518(7540):547-551.
[9]SAHARINEN P,EKLUND L,MIETTINEN J,et al.Angiopoietins assemble distinct Tie2 signalling complexes in endothelial cell-cell and cell-matrix contacts[J].Nat Cell Biol,2008,10(5):527-537.
[10]BARTON W A,DALTON A C,SEEGAR T C M,et al.Tie2 and Eph receptor tyrosine kinase activation and signaling[J].Cold Spring Harb Perspect Biol,2014,6(3):a009142.
[11]MOSS A.The angiopoietin:Tie 2 interaction:a potential target for future therapies in human vascular disease[J].Cytokine Growth Factor Rev,2013,24(6):579-592.
[12]LELIGDOWICZ A,RICHARD-GREENBLATT M,WRIGHT J,et al.Endothelial activation:the Ang/Tie axis in sepsis[J].Front Immunol,2018,9:838.
[13]LIMS S,KOOKSH,LEEJ C.COMP-Ang1 enhances DNA synthesis and cell cycle progression in human periodontal ligament cells via Tie2-mediated phosphorylation of PI3K/Akt and MAPKs[J].Mol Cell Biochem,2016,416(1-2):157-168.
[14]CHITHRANI D B.Intracellular uptake,transport,and processing of gold nanostructures[J].Mol Membr Biol,2010,27(7):299-311.
[15]ALKILANY A M,NAGARIA P K,HEXEL C R,et al.Cellular uptake and cytotoxicity of gold nanorods:molecular origin of cytotoxicity and surface effects[J].Small,2009,5(6):701-708.