体外培养牛眼虹膜色素上皮细胞维生素A代谢相关蛋白mRNA的表达

Expression of mRNA for proteins involved in retinol metabolism in cultured bovine iris pigment epithelial cells in vitro

目的 评价体外培养牛眼虹膜色素上皮 (irispigmentepithelial,IPE)细胞的维生素A代谢功能。方法 采用酶辅助的显微分离法和培养新生牛眼IPE细胞 ,Trizol一步法提取总RNA。根据Genbank中维生素A结合蛋白 (retinol bindingprotein ,RBP)和细胞内视醛结合蛋白 (cellularretinldehyde bindingprotein ,CRALBP)的相关系列设计特异引物 ,对其mRNA的表达进行逆转录聚合酶链反应(reversetranscriptionpolymerase chainreaction ,RT PCR)检测。结果 传 2代牛眼IPE细胞中提取的总RNA完整、无降解。RT PCR检测结果 :RBP及CRALBP的mRNA在培养的牛眼IPE细胞中均有表达。结论 牛眼IPE细胞具有与视网膜色素上皮细胞相似的维生素A转运和代谢功能。

Objective To assess the retinol metabolism of cultured bovine iris pigment epithelial (IPE) cells Methods IPE cells were isolated from neonatal bovines with Hu′s method, and were cultured The total RNA of IPE cells was extracted by Trizol one step method The specific primers for retinol binding protein (RBP) and cellular retinaldehyde binding protein (CRALBP) were designed according to their sequence from Genbank The mRNA expression of these proteins in the IPE cells was analyzed by reverse transcription polymerase chain reaction(RT PCR). Results The extracted RNA from IPE cells of the second passage was ideal and had no degradation RT PCR analysis showed that RBP mRNA and CRALBP mRNA were expressed in cultured IPE cells in vitro Conclusions Similar to RPE cells, cultured IPE cells may have potential of transporting and metabolizing retinol