a Faculty of Veterinary Science , Department of Pharmacology and Toxicology, Szent István University , Budapest , Hungary.
b Faculty of Pharmacy , Semmelweis University , Budapest , Hungary.
J Enzyme Inhib Med Chem. 2016;31(sup2):123-129. doi: 10.1080/14756366.2016.1193732. Epub 2016 Jun 9.
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.
跨膜丝氨酸蛋白酶 TMPRSS2 是治疗季节性流感感染的重要靶点，并且有助于前列腺癌的发生和转移。在这项研究中，研究人员对培养在膜插入物上的肠上皮细胞单层中合成的 TMPRSS2 抑制剂 I-432 的作用进行了表征。研究发现，通过荧光底物，I-432 可抑制 IPEC-J2 细胞上清液中的胰蛋白酶样活性。TMPRSS2 的抑制作用并不影响顶侧和基底外侧隔室中生理产生的过氧化氢水平。当细胞预先暴露于 I-432 时，也观察到 TMPRSS2 丝氨酸蛋白酶结构域（28 kDa）的表达缺失。在用抑制剂孵育 IPEC-J2 细胞 48 小时后，检测到源自 TMPRSS2 分布模式改变的免疫荧光信号强度的部分降低，表明通过 I-432 给药在体外抑制 TMPRSS2 的效果。
