摘要
Introduction Methods Cell lines and culture conditions Exposure of IPEC-J2 cells to TMPRSS2 inhibitor Protein extraction Western blot Protease activity at the cell surface and in cell supernatants Extracellular H2O2measurement by the Amplex red method Investigation of TMPRSS2 distribution via immunfluorescent staining Statistical analysis Results Expression of TMPRSS2 in IPEC-J2 cells Enzymatic activity of TMPRSS2 Extracellular H2O2produced by IPEC-J2 cells exposed to I-432 Subcellular localization of TMPRSS2 in IPEC-J2 cells Discussion Conclusion References type=text/javascript src="/swfobject.js"> type=text/javascript> function addFlashMovie(id, flv) { var flashvars = {file: flv ,type: 'flv'}; var params = {allowfullscreen :true}; var attributes = {}; swfobject.embedSWF('/flvplayer.swf', id, "352", "288", "7.0.0", false, flashvars, params, attributes); } function addFlashMovie(id, flv, image) { var flashvars = {file: flv ,type: 'flv', image: image}; var params = {allowfullscreen :true}; var attributes = {}; swfobject.embedSWF('/flvplayer.swf', id, "352", "288", "7.0.0", false, flashvars, params, attributes); } type=text/javascript> window.MathJax = { AuthorInit: function () { $.getScript( '/templates/jsp/_style2/_tandf/pb2/js/mathJaxConfiguration.js' ); } }; type=text/javascript defer src="http://cdn.mathjax.org/mathjax/latest/MathJax.js?config=TeX-AMS-MML_HTMLorMML" async> Research Article In vitro characterization of TMPRSS2 inhibition in IPEC-J2 cells Full Article Figures & data References Citations Metrics Reprints & Permissions PDF
Abstract
Abstract
The transmembrane serine protease, TMPRSS2 is an important target in the treatment of seasonal influenza infections and contributes to prostate carcinogenesis and metastasis. In this study, the effect of the synthetic TMPRSS2 inhibitor I-432 on jejunal IPEC-J2 cell monolayers cultured on membrane inserts was characterized. Using a fluorogenic substrate, it was found that the apical addition of I-432 could suppress trypsin-like activity in the supernatants of IPEC-J2 cells. The inhibition of TMPRSS2 did not affect physiologically produced hydrogen peroxide levels in the apical and in basolateral compartments. Loss of expression of the TMPRSS2 serine protease domain (28 kDa) was also observed when cells were pre-exposed to I-432. Partial decrease in immunofluorescent signal intensities derived from the altered distribution pattern of TMPRSS2 was detected after a 48 h long incubation of IPEC-J2 cells with the inhibitor indicating the efficacy of TMPRSS2 inhibition via I-432 administration in vitro.
摘要译文
RSS2细胞暴露于I-432的IPEC-J2细胞产生的细胞外H2O2在IPEC-J2细胞中TMPRSS2的亚细胞定位讨论结论参考文献type \x3d text / javascript src \x3d“/swfobject js“ type \x3d text / javascript function addFlashMovie（id，flv）{var flashvars \x3d {file：flv，type：''''flv''''}; var params \x3d {allowfullscreen：true}; var attributes \x3d {}; swfobject竐mbedSWF （''''/ flvplayer竤wf''''，id，“352”，“288”，“7”，false，flashvars，params，attributes）; } function addFlashMovie（id，flv，image）{var flashvars \x3d {file：flv，type：''''flv''''，image：image};var params \x3d {allowfullscreen：true}; var attributes \x3d {}; swfobject竐mbedSWF（''''/ flvplayer竤wf''''，id，“352”，“288”，“7”，false，flashvars，params，attributes）; } type \x3d text / javascriptwindow MathJax \x3d {AuthorInit：function（）{$竒etScript（''''/ templates / jsp / _style2 / _tandf / pb2 / js / mathJaxConfiguration竕s''''）; }}; type \x3d text / javascript defer src \x3d“http：// cdn伫athjax org / mathjax / latest / MathJax？n的TMPRSS2抑制在IPEC-J2细胞中的表达全文参考文献相关文章Metrics全文：PDFTMPRSS2是治疗季节性流感感染的重要靶点，有助于前列腺癌发生和转移。在本研究中，描述了合成的TMPRSS2抑制剂I-432对在膜插入物上培养的空肠IPEC-J2细胞单层的影响。使用荧光底物，发现I-432的顶端添加可以抑制IPEC-J2细胞的上清液中的胰蛋白酶样活性TMPRSS2的抑制不影响生理上产生的过氧化氢水平在顶端和基底外侧室当细胞暴露于I-432时，也观察到TMPRSS2丝氨酸蛋白酶结构域（28Da Da）表达的丧失在IPEC-J2细胞与抑制剂的48小时长的温育之后检测到TMPRSS2的改变的分布模式，表明通过I-432施用体外的TMPRSS2抑制的功效
Erzsebet Pászti-Gere[1];Eszter Czimmermann[2];Gabriella Ujhelyi[3];Peter Balla[4];Alexander Maiwald[5];Torsten Steinmetzer[5]. In vitro characterization of TMPRSS2 inhibition in IPEC-J2 cells[J]. Journal of Enzyme Inhibition and Medicinal Chemistry, 2016,31(Sup2): 123-129