文档介绍：原位杂交法检测视网膜色素上皮细胞炎前因子mRNA的表达
作者:宋宗明惠延年瞿佳王雨生马吉献韩泉洪杜红俊
【关键词】色素上皮细胞
关键词: 色素上皮细胞;视网膜;原位杂交;炎前因子;mRNA
摘要:目的观察视网膜色素上皮细胞(RPE)在炎性介质刺激下炎前因子mRNA的表达. 方法培养的人RPE经IL-1β(10μg・L-1 )和脂多糖(1mg・L-1 )的刺激后,用生物素标记的寡核苷酸探针进行原位杂交,检测RPE炎前因子IL-1β,IL-6,IL-8和TNF-. 结果原位杂交显示RPE细胞质中均呈现浓淡不一的淡蓝色着色,-6mRNA在脂多糖刺激下染色较浅,-1β(10μg・L-1 )和脂多糖(1mg・L-1 )刺激下,RPE表达IL-6mRNA的吸光度分别为108±18和35±14,二者之间有显著性差异(P<). 结论在IL-1β和脂多糖刺激下,人RPE可以表达炎前因子IL-1β,IL-6,IL-8和TNF-α的mRNA.
Keywords:retinal pigment epithelium;retina;in situ hy-bridization;proinflammatory factors;mRNA
Abstract:AIM To determine whether retinal pigment epi-thelium(RPE)expresses proinflammatory factors messenger RNA(mRNA)in cultured RPE cells stimulated by inflam-matory Cultured RPE treated with IL-1β(10μg・L-1 )and haride(1mg・L-1 )was used as presence of mRNA coding for IL-1β,IL-6,IL-8and TNFαwas investigated by in situ hybridiza-tion(ISH)with biotin labeled oligonucleotide analysis was performed to determine staining differences be-tween IL-1β,IL-6,IL-8and TNF-αmRNA positive cells showed light to dark blue staining in RPE cytoplasm after being stimulated with IL-1βand IL-6mRNA showed light staining when RPE was treated with LPS,and greater dosage of haride(LPS)failed to enhance RPE mRNA e