中国当代儿科杂志2017，Vol.19Issue(9)：1014-1019,6.DOI:10.7499/j.issn.1008-8830.2017.09.016
戊二酰辅酶A脱氢酶基因沉默及高浓度赖氨酸对BRL肝细胞活性的影响
Effect of glutaryl-CoA dehydrogenase gene silencing and high-concentration lysine on the viability of BRL hepatocytes
高金枝 1张偲 1易琴 1应艳琴 1罗小平1
作者信息
- 1. 华中科技大学同济医学院附属同济医院儿科,湖北 武汉 430030
- 折叠
摘要
Abstract
Objective To investigate the effect of glutaryl-CoA dehydrogenase (GCDH) gene silencing and accumulation of lysine metabolites on the viability of hepatocytes. Methods BRL cells were divided into normal control group, negative control group, and GCDH silencing group. The shRNA lentiviral vector for silencing GCDH gene was constructed, and the BRL hepatocytes in the GCDH silencing group and the negative control group were infected with this lentivirus and negative control virus respectively, and then cultured in a medium containing 5 mmol/L lysine. Immunofluorescence assay was used to measure the infection efficiency of lentivirus. Western blot was used to measure the expression of GCDH protein. MTT assay was used to evaluate cell viability. Hoechest33342 staining was used to measure cell apoptosis. Western blot was used to measure the expression of Caspase-3, an index of cell apoptosis. Results The lentivirus constructed effectively silenced the GCDH gene in hepatocytes (P<0.01). MTT assay and Hoechest 33342 staining showed no significant differences in cell viability and apoptosis between groups (P>0.05). There was also no significant difference in the expression of Caspase-3 protein between groups (P>0.05). Conclusions GCDH gene silencing and accumulation of lysine metabolites may not cause marked hepatocyte injury.关键词
戊二酰辅酶A脱氢酶/细胞活性/BRL细胞Key words
Glutaryl-CoA dehydrogenase/Cell viability/BRL cell引用本文复制引用
高金枝,张偲,易琴,应艳琴,罗小平..戊二酰辅酶A脱氢酶基因沉默及高浓度赖氨酸对BRL肝细胞活性的影响[J].中国当代儿科杂志,2017,19(9):1014-1019,6.
