IB突变型基因对永生化神经前体细胞生物学特性的影....docVIP

IκBα突变型基因对永生化神经前体细胞生物学特性的影响 430030 祝畅( 刘志恒 张传汉 桂伶俐 姚文龙 【摘要】目的 观察IκBα突变型基因对永生化神经前体细胞株（INPCs）增殖、分化及部分炎性细胞因子分泌的影响。方法 在已经建立的转染pcDNA3.1及转染pcDNA3.1/IκBαM的INPCs的基础上，用MTT法绘制两细胞株的生长曲线，用流式细胞仪检测其增殖指数，以5％小牛血清诱导分化后，采用western blot检测两株细胞的分化情况，同时用realtimePCR比较TNF-α、IL-1β和IL-6在两种细胞株中的表达。 结果 生长曲线及流式细胞仪测得的细胞增殖指数提示：转染pcDNA3.1/IκBαM INPCs的增殖速度较转染pcDNA3.1的INPCs明显降低。经血清诱导分化后，两者均大量表达胶原纤维酸性蛋白；转pcDNA3.1/IκBαM的INPCs较转pcDNA3.1的INPCs IL-1β和IL-6的表达减低，但TNF-α表达变化无显著性。结论 IκBα突变型基因可减慢INPCs的增殖，并减少部分炎性细胞因子的表达，但对INPCs的分化无显著影响。 【关键词】干细胞； NF-κB抑制因子α；细胞增殖；细胞分化；细胞因子 Effect of mutated IκBα gene on the biological characteristics of immortalized neural progenitor cell strain ZHU Chang, LIU Zhi-heng, ZHANG Chuan-han, et al. Department of Anesthesiology，Tongji Hospital of Tongji Medical College, Huazhong University of Science Technology, Wuhan 430030, China 【Abstract】 Objective To observe the effect of mutated IκBα gene on proliferation，differentiation and cytokine secretion of immortalized neural progenitor cell strain(INPCs). Methods After the establishment of mutated IκBα gene transfected INPCs and blank plasmid transfected INPCs, MTT staining and flow cytometry used to draw the growth curve and measure the cell proliferation index. Western blot was carried out to observe the differentiation of the two cell strains after 5% blood serum induced differentiation. The expression of TNF-α、IL-1β and IL-6 was measured by realtime RT-PCR. Results It was established by growth curve and cell proliferation index that the proliferation was slowed down in mutated IκBα gene transfected INPCs compared with blank plasmid transfected INPCs. After blood serum induced differentiation，glial fibrillary acidic protein was highly expressed in the two cell strains. There was no difference in the expression of TNF-α between mutated IκBα gene transfected INPCs and blank plasmid transfected INPCs. The expression of IL-1β and IL-6 was reduced in mutated IκBα gene transfected INPCs. Conclusion Mutated IκBα gene can slow down the