国际眼科杂志
國際眼科雜誌
국제안과잡지
INTERNATIONAL JOURNAL OF OPHTHALMOLOGY
2007年
2期
298-300
,共3页
陈立忠%洪晶%海老原 伸行%村上晶
陳立忠%洪晶%海老原 伸行%村上晶
진립충%홍정%해로원 신행%촌상정
连接粘附分子-1%检测咬合蛋白%紧密连接
連接粘附分子-1%檢測咬閤蛋白%緊密連接
련접점부분자-1%검측교합단백%긴밀련접
junction adhesion molecule-1%occludin%tight jundion
目的:检测连接粘附分子-1(junction adhesion molecule,JAM-1)在正常人角膜上皮各层中的表达及分布特点并与咬合蛋白(occludin)进行比较.方法:培养人角膜上皮细胞提取细胞总RNA.以逆转录后获得的cDNA为模板PCR扩增目的基因JAM-1及occludin.流式细胞仪检测JAM-1蛋白表达.双重免疫荧光观察JAM-1与occludin在正常人角膜上皮组织的原位表达.结果:通过RT-PCR在培养人角膜上皮细胞中检测到JAM-1与occludin扩增片段;流式细胞仪检测到JAM-1蛋白表达;双重免疫荧光结果显示occludin染色主要位于表层上皮层细胞之间;而JAM-1荧光染色不仅见于上皮表层,在整个上皮层细胞之间均可见其荧光反应.结论:Occludin主要位于正常人角膜上皮表层细胞之间,JAM-1在正常人角膜上皮的全层中均有表达.
目的:檢測連接粘附分子-1(junction adhesion molecule,JAM-1)在正常人角膜上皮各層中的錶達及分佈特點併與咬閤蛋白(occludin)進行比較.方法:培養人角膜上皮細胞提取細胞總RNA.以逆轉錄後穫得的cDNA為模闆PCR擴增目的基因JAM-1及occludin.流式細胞儀檢測JAM-1蛋白錶達.雙重免疫熒光觀察JAM-1與occludin在正常人角膜上皮組織的原位錶達.結果:通過RT-PCR在培養人角膜上皮細胞中檢測到JAM-1與occludin擴增片段;流式細胞儀檢測到JAM-1蛋白錶達;雙重免疫熒光結果顯示occludin染色主要位于錶層上皮層細胞之間;而JAM-1熒光染色不僅見于上皮錶層,在整箇上皮層細胞之間均可見其熒光反應.結論:Occludin主要位于正常人角膜上皮錶層細胞之間,JAM-1在正常人角膜上皮的全層中均有錶達.
목적:검측련접점부분자-1(junction adhesion molecule,JAM-1)재정상인각막상피각층중적표체급분포특점병여교합단백(occludin)진행비교.방법:배양인각막상피세포제취세포총RNA.이역전록후획득적cDNA위모판PCR확증목적기인JAM-1급occludin.류식세포의검측JAM-1단백표체.쌍중면역형광관찰JAM-1여occludin재정상인각막상피조직적원위표체.결과:통과RT-PCR재배양인각막상피세포중검측도JAM-1여occludin확증편단;류식세포의검측도JAM-1단백표체;쌍중면역형광결과현시occludin염색주요위우표층상피층세포지간;이JAM-1형광염색불부견우상피표층,재정개상피층세포지간균가견기형광반응.결론:Occludin주요위우정상인각막상피표층세포지간,JAM-1재정상인각막상피적전층중균유표체.
AIM:To investigate the expression and distribution of junction adhesion molecule-1(JAM-1)in human corneal epithelium and compare with that of occludin.METHODS:The expression in RNAs of JAM-1 and occludin was revealed by RT-PCR and the presence of protein was analyzed by the FACS method.Double immunofluorescent staining was used to determine the tissue distribution of JSM-1 and occludin in human corneal epithelium.RESULTS:The expression of JAM-1 and occludin was found in cultured human corneal epithelial cells.The double immunofluorescent study showed positive staining for JAM-1 at cell borders in the entire epithelial layer,while relatively extensive staining was seen in the superficial layer,where it COexisted with the expression of ocdudin.CONCLUSION:JAM-1 was expressed in entire layer of human corneal epithelium encircling the cells.
