Utami Ratna Annisa, Hakiki Aunillah, Asyarie Sukmadjaja, Retnoningrum Debbie Soefie
Laboratory of Pharmaceutical Biotechnology, School of Pharmacy, Institut Teknologi Bandung, Jalan Ganesha No. 10 Bandung, West Java 40132, Indonesia.
Sci Pharm. 2018 Apr 10;86(2):13. doi: 10.3390/scipharm86020013.
Superoxide dismutase (SOD) is an antioxidant protein. When administered orally, it has low bioavailability due to its low permeation. In a previous study we fused gliadin peptide P51 (LGQQQPFPPQQPYPQPQPF) and gliadin peptide P61 (QQPYPQPQPF) with SOD (SOD_Cl), namely GliSOD_P51 and GliSOD_P61 to increase permeation of SOD_Cl through intestine. In this work, the permeation of fluorescein isothiocyanate (FITC)-Dextran 10 kDa, FD10 and 40 kDa, FD40 as paracellular transport markers across excised rat intestinal wall was investigated with the presence of GliSOD_P51 and GliSOD_P61. A permeability study was performed using non-everted rat intestine by incubating FD10 or FD40 with SOD_Cl, and GliSOD_P61. The presence of SOD_Cl, GliSOD_P51 or GliSOD_P61 inside intestine (apical) and outside intestine (basolateral) was analyzed by protein electrophoresis. The concentration of FD that penetrated to the basolateral solution was analyzed by spectrofluorometry. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed the presence of GliSOD_P51 and GliSOD_P61 but not SOD_Cl in basolateral compartment. The percentage of FD10 but not FD40 and SOD_Cl that penetrated to the basolateral solution significantly increased with the presence of gliadin in GliSOD_P51 and GliSOD_P61. GliSOD_P51 and GliSOD_P61 are able to penetrate the rat intestinal epithelial membrane and the gliadin peptides facilitate FD10 to penetrate the epithelial.
超氧化物歧化酶（SOD）是一种抗氧化蛋白。口服时，由于其低渗透性，生物利用度较低。在之前的一项研究中，我们将麦醇溶蛋白肽P51（LGQQQPFPPQQPYPQPQPF）和麦醇溶蛋白肽P61（QQPYPQPQPF）与SOD（SOD_Cl）融合，即GliSOD_P51和GliSOD_P61，以增加SOD_Cl通过肠道的渗透性。在这项工作中，研究了异硫氰酸荧光素（FITC）-葡聚糖10 kDa（FD10）和40 kDa（FD40）作为细胞旁转运标志物在GliSOD_P51和GliSOD_P61存在的情况下穿过离体大鼠肠壁的渗透性。使用未翻转的大鼠肠道进行通透性研究，将FD10或FD40与SOD_Cl和GliSOD_P61一起孵育。通过蛋白质电泳分析肠道内（顶端）和肠道外（基底外侧）SOD_Cl、GliSOD_P51或GliSOD_P61的存在情况。通过荧光分光光度法分析渗透到基底外侧溶液中的FD浓度。十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）分析显示基底外侧隔室中存在GliSOD_P51和GliSOD_P61，但不存在SOD_Cl。在GliSOD_P51和GliSOD_P61中存在麦醇溶蛋白的情况下，渗透到基底外侧溶液中的FD10的百分比显著增加，而FD40和SOD_Cl则没有。GliSOD_P51和GliSOD_P61能够穿透大鼠肠上皮膜，并且麦醇溶蛋白肽促进FD10穿透上皮。
