应用HIOEC细胞单层模型研究胰岛素在口腔上皮的渗透特性

目的用口腔永生化上皮(human immortalized oral epithelial cell,HIOEC)细胞建立Transwell单细胞层转运模型,研究不同条件和不同渗透促进剂、不同转运添加剂对胰岛素在口腔上皮细胞渗透转运中作用的影响。方法以胰岛素为模型药,测定不同条件下(时间、温度、转运方向)、不同渗透促进剂(壳聚糖、蛋黄卵磷脂、Brij35等)、不同转运添加剂(秋水仙碱、多聚-L-氨酸)作用下胰岛素在HIOEC单细胞层的表观渗透系数Papp,观察胰岛素的渗透转运特性。结果1)胰岛素在HIOEC单细胞层的转运不具有浓度、温度、转运方向依赖性。2)渗透促进剂Brij35、蛋黄卵磷脂和壳聚糖均能显著增加胰岛素在HIOEC单细胞层的渗透转运,其通过细胞间紧密连接破坏膜脂质的有序性,改变插入到细胞间蛋白的构象和位置,从而增加单细胞层脂质流动性与细胞间紧密连接的空间,促进胰岛素的渗透转运。3)阳离子氨基酸多聚-L-赖氨酸通过与膜表面的阴离子结合破坏细胞膜脂质性质,打开细胞间紧密连接,增加胰岛素在HIOEC单细胞层的渗透转运。结论渗透促进剂可通过增加膜脂流动性、改变膜蛋白构象和位置、打开HIOEC细胞间紧密连接等方式提高胰岛素在HIOEC单细胞层的渗透转运。

Model and Mechanism of the Insulin Transport across Human Immortalized Oral Epithelial Cell Monolayers

Objective The purposes of this study are to evaluate the mechanism of delivering insulin with enhancer and additives on the in vitro cultured monolayer system of HIOEC(the human immortalized oral epithelial cell) lines.Methods The HIOECs formed continuous monolayers with polycarbonate filters of Transwell plate served to evaluate the transport of insulin.The transport studies were carried out at different conditions,with different penetration enhancers(HP-β-CD,chitosan,polyoxyethylene lauryl ether,egg lecithin,or oleic acid),and different additives(poly-L-lysine,colchicine).Results Under different conditions,the transport of insulin was not direction-,temperature-,and concentration-dependent.Conclusion The enhancing effects might be due to one or multiple factors:increasing the mucosal lipid fluidity,directly loosing the tight junction,and changing the steric conformation of protein.