同源盒基因Lhx4在PC12细胞缺氧损伤保护中的作用

目的:转染并筛选Lhx4高表达的PC12细胞株,观察Lhx4基因对PC12细胞的缺氧保护作用.方法:①用RT-PCR检测PC12细胞缺氧培养后Lhx4基因的表达变化;②Lhx4基因连接至pLXIN逆转录病毒载体,收集病毒上清,感染PC12细胞,G418筛选阳性克隆,PCR鉴定,所筛选的克隆缺氧培养;③通过台盼蓝染色和培养液中乳酸脱氢酶活性的检测评价PC12细胞的存活状态.结果和结论:PC12细胞缺氧培养4 h后Lhx4基因的表达明显上调;缺氧8 h后Lhx4基因表达开始下调.所筛选的Lhx4高表达转染细胞株表现出明显的缺氧耐受性,相同的损伤条件下,细胞的存活数目及生存状态明显优于对照组.说明Lhx4基因在PC12细胞的缺氧损伤保护中具有重要的作用.

Protective effect of human homeobox gene-Lhx4 against PC12 cells injury induced by anoxia

Objective:To observe the protective effect of Lhx4 gene against the PC12 cells injury induced by anoxia.Methods:(1) The LHX4 CDS fragment was ligated to the retroviral vector pLXIN,and transfected into PC12 cells,then LHX4-pLXIN PC12 cell clones were selected using G418.(2) The expression of Lhx4 gene in PC12 cells under anoxia was tested by RT-PCR.(3) The survival rate of PC12 cells was measured by Trypan blue staining and the activity of LDH in culture medium.Results: The expression of Lhx4 gene increased when PC12 cells were exposed to anoxia condition for 4 hours and decreased after PC12 cells were exposed to anoxia for 8 hours.Overexpression of Lhx4 in the PC12 cells could significantly increase the survival of PC12 cells injured by anoxia.Conclusion: The results indicate that Lhx4 gene plays a protective role against PC12 cell injury under anoxia.