解放军医学院学报Issue(8)：826-828,3.DOI:10.3969/j.issn.2095-5227.2015.08.017
组织金属蛋白酶抑制剂-1低表达系膜细胞模型的建立
Establishment of TIMP1 low-expression model in mesangial cells
摘要
Abstract
Objective To compound three different small interference RNAs specific for TIMP-1 and establish low-expression model of TIMP-1 in mesangial cells of rats, in order to provide experimental basis for renal diseases.MethodsThree different small interference RNA sequences that were specific for TIMP-1 were designed and compounded. The nonsense siRNA marked with greenfluorescent perssad (6-carboxy-fluorescein, FAM) was used as control group and the three siRNA groups were transfected into cultured mesangial cells in rats by liposome transfection method, then the transfected cells were collected at 48 hours after transfection to identify the transfection efficiency withfluorescent microscope. At the same time, the total RNA in mesangial cells was extracted with Trizol, the TIMP-1 mRNA expression level of all four groups was detected by Taqman probe technique.Results Thefluorescent microscope revealed that the transfection efficiency was over than 90% in nonsense siRNA group, while the PCR results demonstrated that TIMP-1 mRNA expression level in all three groups was lower than control group (P<0.01), which suggested that our three siRNAs had inhibited the expression of TIMP-1, and siTIMP1-1 had the best effect for it showed the lowest expression level.Conclusion Our study has successfully established the low-expression cell model of TIMP-1 in primary MCs with siRNA transfection technique.关键词
组织金属蛋白酶抑制剂-1/小干扰RNA/系膜细胞Key words
tissue inhibitors of metalloproteinase-1/siRNA/mesangial cells分类
医药卫生引用本文复制引用
陈曦曌,吕杨,谢院生,陈香美..组织金属蛋白酶抑制剂-1低表达系膜细胞模型的建立[J].解放军医学院学报,2015,(8):826-828,3.基金项目
国家自然科学基金项目(81330019；81270794)；国家重大科学研究计划项目(2011CB944000)Supported by the National Natural Science Foundation of China(81330019 （81330019；81270794）
81270794) （）
National Science and Technology Major Project(2011CB944000) （2011CB944000）
