EVALUASI METODE USUS TIKUS TIDAK DIBALIK UNTUK UJI PERMEABILITAS MENGGUNAKAN SUPEROKSIDA DISMUTASE Citrus limon –GLIADIN SEBAGAI MODEL PROTEIN

• Main Author: Hakiki, Aunillah
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/44268
• Institution: Institut Teknologi Bandung

To increase the permeability of superoksida dismutase Citrus limon (SOD_Cl), in previous research superoxide dismutase Citrus limon (SOD_Cl) was fused to gliadin peptide as permeation enhancer. The results of permeability test of SOD_Cl and its fusion forms with gliadin peptides LGQQQPFPPQQPYPQPQPF (GliSOD_P51) and QQPYPQPQPF (GliSOD_P61) using Caco-2 cell culture (21 days old) showed that both gliadin peptides increased the permeability of SOD_Cl and Fluorescein isothiocyanate-Dextran (FD) 10 kDa but not FD 40 kDa. In this study, evaluation of permeability testing performance was examined using non everted rat intestine with SOD_Cl, GliSOD_P51 and GliSOD_P61 as protein model as well as FD 10 kDa and 40 kDa as paracellular transport markers. Research was initiated by overexpression and purification SOD_Cl, GliSOD_P51, and GliSOD_P61. Permeability study was performed using non everted rat intestine by incubating FD 10 kDa or 40 kDa with the three proteins. The presence of three proteins inside intestine (apical) and outside intestine (basolateral) were analyzed by protein electrophoresis and zymography, while the concentration of FD that was penetrated to basolateral solution was analyzed by spectrofluorometry. The protein electrophoresis results showed the presence of GliSOD_P51 and GliSOD_P61 in basolateral solution, while not SOD_Cl. The concentrations of FD 10 kDa and 40 kDa on basolateral solution without protein were higher than those with proteins. Percentage of FD 10 kDa penetrated to basolateral solution significantly increased with the presence of gliadin on GliSOD_P51 and GliSOD_P61 compared to SOD_Cl while for 40 kDa concentrations there was no significant difference. The result of permeability test using non everted rat intestine has low correlation with Caco-2 cell culture 21 day.