经角膜途径是药物进入眼内部位的主要途径,局部给药后。外表面,角膜上皮(CE),形成药物渗透性的限速屏障。关于药物和氨基酸转运蛋白在CE中的作用和蛋白质表达的信息很少且缺乏。我们研究的目的是表征兔和猪CE中转运蛋白的表达,以更好地了解局部给药后潜在的药物和营养吸收。蛋白质,主要是Abc和Slc转运蛋白,用定量靶向绝对蛋白质组学和全局非靶向蛋白质组学方法进行表征。在兔子CE中,在靶向方法中检测到48种蛋白质中的24种,其中21个被量化。在猪CE中,在靶向方法中检测到58种蛋白质中的26种,其中20个被量化。其中,在两只动物中定量了15种蛋白质:4f2hc(Slc3a2),Aqp0,Asct1(Slc1a4),Asct2(Slc1a5),Glut1(Slc2a1),Hmit(Slc2a13),Insr,Lat1(Slc7a5),Mct1(Slc16a1),Mct2(Slc16a7),Mct4(Slc16a3),Mrp4(Abcc4),Na+/K+-ATP酶,Oatp3a1(Slco3a1),和Snat2(Slc38a2)。总的来说,全球蛋白质组学结果支持靶向蛋白质组学结果.在兔(1.4±0.4fmol/cm2)和猪(11.1±5.3fmol/cm2)CE中首次检测并定量了有机阴离子转运多肽Oatp3a1。观察到L型氨基酸转运蛋白的高表达水平,Lat1,其在兔(48.9±11.8fmol/cm2)和猪(37.6±11.5fmol/cm2)CE中用新选择的胞外结构域肽定量。了解转运体蛋白在眼部屏障中的表达是成功设计新型眼部药物的关键因素。药代动力学建模,了解眼部疾病,以及对人类的翻译。
The transcorneal route is the main entry route for drugs to the intraocular parts, after topical administration. The outer surface, the corneal epithelium (CE), forms the rate-limiting barrier for drug permeability. Information about the role and protein expression of drug and amino acid transporter proteins in the CE is sparse and lacking. The aim of our study was to characterize transporter protein expression in rabbit and porcine CE to better understand potential drug and nutrient absorption after topical administration. Proteins, mainly Abc and Slc transporters, were characterized with quantitative targeted absolute proteomics and global untargeted proteomics methods. In the rabbit CE, 24 of 48 proteins were detected in the targeted approach, and 21 of these were quantified. In the porcine CE, 26 of 58 proteins were detected in the targeted approach, and 20 of these were quantified. Among these, 15 proteins were quantified in both animals: 4f2hc (Slc3a2), Aqp0, Asct1 (Slc1a4), Asct2 (Slc1a5), Glut1 (Slc2a1), Hmit (Slc2a13), Insr, Lat1 (Slc7a5), Mct1 (Slc16a1), Mct2 (Slc16a7), Mct4 (Slc16a3), Mrp 4 (Abcc4), Na+/K+-ATPase, Oatp3a1 (Slco3a1), and Snat2 (Slc38a2). Overall, the global proteomics results supported the targeted proteomics results. Organic anion transporting polypeptide Oatp3a1 was detected and quantified for the first time in both rabbit (1.4 ± 0.4 fmol/cm2) and porcine (11.1 ± 5.3 fmol/cm2) CE. High expression levels were observed for L-type amino acid transporter, Lat1, which was quantified with newly selected extracellular domain peptides in rabbit (48.9 ± 11.8 fmol/cm2) and porcine (37.6 ± 11.5 fmol/cm2) CE. The knowledge of transporter protein expression in ocular barriers is a key factor in the successful design of new ocular drugs, pharmacokinetic modeling, understanding ocular diseases, and the translation to human.