华中科技大学学报（医学版）
華中科技大學學報（醫學版）
화중과기대학학보（의학판）
ACTA UNIVERSITATIS MEDICINAE TONGJI
2014年
6期
643-647
,共5页
安翠平%毕建成%李宝全%许顺江%宋国威
安翠平%畢建成%李寶全%許順江%宋國威
안취평%필건성%리보전%허순강%송국위
肉碱/有机阳离子转运体%角膜%结膜%上皮细胞%转运
肉堿/有機暘離子轉運體%角膜%結膜%上皮細胞%轉運
육감/유궤양리자전운체%각막%결막%상피세포%전운
carnitine/organic cation transporter%cornea%conjunctiva%epithelia cells%transport
目的：检测肉碱／有机阳离子转运体1／2（carnitine／organic cation transporter 1／2，OCTN1／2）在眼表上皮细胞的表达定位及转运功能，为进一步阐明左旋肉碱在人眼表上皮细胞的转运机制提供实验依据。方法采用免疫细胞化学的方法分别检测人角膜缘上皮（human corneal limbal epithelia ，HCLE）细胞和人结膜上皮（human conjunctival epithe‐lia ，HCjE）细胞以及兔眼角膜和结膜上皮组织中OCTN1和OCTN2的表达定位；利用Ⅰ型胶原蛋白包被的双层培养板培养HCLE和 HCjE细胞，并使之建立紧密连结，采用放射摄入实验检测其基底端和顶端转运[3 H]‐L‐carnitine的功能。结果 OCTN1和OCTN2蛋白在HCLE和HCjE细胞及兔眼角膜和结膜上皮组织中均有表达，且主要分布在细胞膜上；放射摄入实验结果表明 HCLE和HCjE细胞均可转运[3 H]‐L‐carnitine ，且大部分的左旋肉碱分子通过上皮细胞顶端摄入细胞内。结论 OCTN1和OCTN2主要分布在人角膜和结膜上皮细胞的顶端，且具有转运左旋肉碱进入细胞内的功能。
目的：檢測肉堿／有機暘離子轉運體1／2（carnitine／organic cation transporter 1／2，OCTN1／2）在眼錶上皮細胞的錶達定位及轉運功能，為進一步闡明左鏇肉堿在人眼錶上皮細胞的轉運機製提供實驗依據。方法採用免疫細胞化學的方法分彆檢測人角膜緣上皮（human corneal limbal epithelia ，HCLE）細胞和人結膜上皮（human conjunctival epithe‐lia ，HCjE）細胞以及兔眼角膜和結膜上皮組織中OCTN1和OCTN2的錶達定位；利用Ⅰ型膠原蛋白包被的雙層培養闆培養HCLE和 HCjE細胞，併使之建立緊密連結，採用放射攝入實驗檢測其基底耑和頂耑轉運[3 H]‐L‐carnitine的功能。結果 OCTN1和OCTN2蛋白在HCLE和HCjE細胞及兔眼角膜和結膜上皮組織中均有錶達，且主要分佈在細胞膜上；放射攝入實驗結果錶明 HCLE和HCjE細胞均可轉運[3 H]‐L‐carnitine ，且大部分的左鏇肉堿分子通過上皮細胞頂耑攝入細胞內。結論 OCTN1和OCTN2主要分佈在人角膜和結膜上皮細胞的頂耑，且具有轉運左鏇肉堿進入細胞內的功能。
목적：검측육감／유궤양리자전운체1／2（carnitine／organic cation transporter 1／2，OCTN1／2）재안표상피세포적표체정위급전운공능，위진일보천명좌선육감재인안표상피세포적전운궤제제공실험의거。방법채용면역세포화학적방법분별검측인각막연상피（human corneal limbal epithelia ，HCLE）세포화인결막상피（human conjunctival epithe‐lia ，HCjE）세포이급토안각막화결막상피조직중OCTN1화OCTN2적표체정위；이용Ⅰ형효원단백포피적쌍층배양판배양HCLE화 HCjE세포，병사지건립긴밀련결，채용방사섭입실험검측기기저단화정단전운[3 H]‐L‐carnitine적공능。결과 OCTN1화OCTN2단백재HCLE화HCjE세포급토안각막화결막상피조직중균유표체，차주요분포재세포막상；방사섭입실험결과표명 HCLE화HCjE세포균가전운[3 H]‐L‐carnitine ，차대부분적좌선육감분자통과상피세포정단섭입세포내。결론 OCTN1화OCTN2주요분포재인각막화결막상피세포적정단，차구유전운좌선육감진입세포내적공능。
Objective To investigate the localization and transport function of carnitine/organic cation transporter 1/2(OC‐TN1/2) in ocular epithelium ,and to provide a structural and functional basis for further study of transport mechanism of L‐carnitine in human ocular epithelium.Methods Immunofluorescence staining was performed to investigate the localization of OCTN1 or OCTN2 in human ocular epithelial cells or rabbit corneal and conjunctival epithelium.Apical or basal uptake of [3 H]‐L‐carnitine was determined by using the polarized epithelial cells grown onto collagen‐coated porous filter support.Results OCTN1 and OCTN2 proteins were detected in HCLE and HCjE cells ,rabbit corneal and conjunctival epithelium ,and they were predominately located in the membranes of the cells. HCLE and HCjE cells were able to transfer [3 H]‐L‐carnitine and most L‐carnitine was uptaken through the apical part of the cells.Conclusion OCTN1 and OCTN2 are mainly located in the apical part of human corneal and conjunctival epithelial cells ,and they are potentially involved in the transport of L‐carnitine in ocular tis‐sues.
