山东医药Issue(7)：17-19,3.DOI:10.3969/j.issn.1002-266X.2016.07.006
Prx-1基因沉默对 TGF-β1诱导肺成纤维细胞增殖、ROS 水平、p-AKT 蛋白表达的影响
Effects of silencing peroxiredoxin-1 gene on proliferation, ROS and p-AKT protein expression of pulmonary fibroblasts induced by TGF-β1
摘要
Abstract
Objective To investigate the effects of silencing peroxiredoxin-1 (Prx-1) gene on the proliferation, reac-tive oxygen species ( ROS) and phosphorylated AKT ( p-AKT) protein expression of pulmonary fibroblasts induced by TGF-β1 .Methods The cultured MRC-5 pulmonary fibroblasts were randomly divided into 4 groups: control group, TGF-β1 group, negative transfection group and experimental group.The Prx-1 siRNA 3 sequences of Prx-1 siRNA ( Prx-1 siRNA-209, Prx-1 siRNA-289, Prx-1 siRNA-453) were transfected into the negative transfection group and experimental group by Lipofectamine2000, respectively.After culturing for 48 h, Prx-1 mRNA was detected by real-time PCR to evaluate trans-fection and Prx-1 siRNA-453 was selected to be used later.Except for the control group, the other three groups were co-cultured with TGF-β1(5 μg/L).Cell proliferation was detected by MTT assay and reactive oxygen species (ROS) level was detected by DCFH-DA assay after 24 h of TGF-β1 stimulation.p-AKT expression was detected by Western blotting after 45 min of TGF-β1 stimulation.Results The OD was respectively 0.56 ±0.07, 0.81 ±0.10, 0.88 ±0.18 and 1.16 ± 0.18 in the control group, TGF-β1 group, negative transfection group and experimental group, and the levels of ROS were 2 922 ±291, 4 348 ±484, 4 660 ±375 and 5 415 ±436, respectively.Significant difference were found in the levels of ROS and OD between the experimental group and the other three groups (all P<0.01).The p-AKT expression of the con-trol group, TGF-β1 group, negative transfection group and experimental group was 0.45 ±0.05, 0.60 ±0.07, 0.57 ±0.07 and 0.77 ±0.09, respectively.The expression of p-AKT in the experimental group was the highest in all groups (all P<0.01).The p-AKT expression of the TGF-β1 group was higher than that of the control group (P<0.01).Conclusion Silencing Prx-1 gene can enhance the proliferation of pulmonary fibroblasts induced by TGF-β1 , and up-regulate the level of ROS and p-AKT protein expression in the cells.关键词
肺成纤维细胞/硫氧还原蛋白过氧化物酶1/基因沉默/转化生长因子β1/细胞增殖/活性氧簇/蛋白激酶BKey words
lung fibroblasts/peroxiredoxin/gene Silencing/transforming growth factro-β1/cell proliferation/reac-tive oxygen species/protein kinase B分类
医药卫生引用本文复制引用
刘宝欣,刘英宇,魏中秋,梁婷婷,范玉磊,杨方,孙影..Prx-1基因沉默对 TGF-β1诱导肺成纤维细胞增殖、ROS 水平、p-AKT 蛋白表达的影响[J].山东医药,2016,(7):17-19,3.基金项目
国家自然科学基金资助项目（81072254）；唐山市科学技术研究与发展项目（14130275B）。 （）
