关键词: 吗啉代寡核苷酸技术, lmna基因, 凋亡蛋白, 流式细胞术, western blot免疫蛋白印迹

Abstract: Objective The mechanism of apoptosis of zebrafish embryo cells after lmna was down regulated by morpholine oligonucleotides. Methods Select the appropriate target, design for lmna morpholino oligonucleotides sequence (lmna-MO),microinjection of lmna-MO into wild type zebrafish embryos to down regulate LMNA. Detection of cell apoptosis by flow cytometry and detection of apoptotic protein p-AKT,AKT and BCL-2 expression by Western blot.Results Early apoptosis of 24 h and 72 h zebrafish embryos was observed after administration of lmna-MO, There was correlation with the phase of drug exposure(p＜0.01); The expression of p-AKT protein is higher than that of wild type embryos(p＜0.01);The expression of AKT was increased compared with wild type embryo cells(p＜0.05), the expression of BCL-2 protein in embryo cells decreased after lmna-MO injection of 24 h and 72 h compared with wild type embryo cells(p＜0.05).Conclusions lmna may be involved in the regulation of zebrafish embryonic cell apoptosis, and its mechanism may be related to the expression of AKT protein and BCL-2 protein; p-AKT（S473）participates in the activation of AKT pathway. Down regulation of lmna may be mediated by the promotion of phosphorylation of AKT，activation of P13K/AKT signaling pathway inhibits apoptosis and promotes cell survival, but it is not the only regulatory mechanism.

Key words: morpholine oligonucleotide, LMNA gene, apoptosis protein, flow cytometry, western blot