摘要
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
摘要跨膜丝氨酸蛋白酶TMPRSS2是治疗季节性流感感染的重要靶点，有助于前列腺癌的发生和转移。在该研究中，表征了合成的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的抑制不影响顶端和基底外侧隔室中生理上产生的过氧化氢水平。当细胞预暴露于I-432时，也观察到TMPRSS2丝氨酸蛋白酶结构域（28kDa）的表达缺失。在IPEC-J2细胞与抑制剂一起长时间孵育48小时后，检测到源自改变的TMPRSS2分布模式的免疫荧光信号强度的部分降低，所述抑制剂通过体外I-432施用表明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, 2016,31(Sup2): 123-129