山东医药2017，Vol.57Issue(34)：5-8,4.DOI:10.3969/j.issn.1002-266X.2017.34.002
小干扰RNA与反义寡核苷酸技术抑制肝星状细胞RhoA表达的效果比较
Effect comparison of small interfering RNA and antisense oligonucleotide in inhibition of RhoA expression of rat hepatic stellate cells
摘要
Abstract
Objective To compare the effects of small interfering RNA (siRNA) and antisense oligonucleotide (ASODN) in inhibition of RhoA expression of rat hepatic stellate cells.Methods Rat hepatic stellate cell line HSC-T6 was divided into the siRNA group and ASODN group, respectively, and then each group was separately divided into subgroups A, B, C, and D.Cells in the subgroup A of the siRNA group and ASODN group were cultured normally, subgroup B was transfected with HK-A negative control plasmid, subgroup C with Rat1, and subgroup D with Rat2.After transfection for 48 h, the mRNA expression of RhoA and ColⅠin HSC-T6 cells of each group was detected by reverse-transcription polymerase chain reaction (RT-PCR), respectively;the content of hyaluronic acid (HA) and laminin (LN) in culture serum was measured by specific ELISA.Results The expression of RhoA mRNA and Col I mRNA in the subgroup C of siRNA group and ASODN group was the lowest, and significant difference was found between subgroup A and subgroup C (both P<0.05), but no significant difference was found between the rest of subgroups (all P>0.05).The mRNA expression of RhoA and Col Ⅰ in the subgroup C of the siRNA group was lower than that in the ASODN group (P<0.05).There was no significant difference in the subgroups A, B, and D between the siRNA group and ASODN group (all P>0.05).The levels of HA and LN in the supernate of subgroup C of the siRNA group and ASODN group were the lowest, and significant difference was found between subgroup A and subgroup C (both P<0.05), but no significant difference was found between the rest of subgroups (all P>0.05).The levels of HA and LN in the supernate of subgroup C of the siRNA group were lower than those in the ASODN group (both P<0.05).There was no significant difference in the subgroups A, B, and D between the siRNA group and ASODN group (all P>0.05).Conclusion Both siRNA and ASODN can inhibit RhoA expression of rat HSC-T6 cells, and RNA interference mediated by Rat1 plasmids targeting RhoA can more effectively inhibit the formation of extracellular matrix HA, Col Ⅰ, and LN than that of ASODN.关键词
小干扰RNA/反义寡核苷酸/RhoA/透明质酸/Ⅰ型胶原/层粘连蛋白/肝星状细胞Key words
small interfering RNA/antisense oligonucleotide/RhoA/hyaluronic acid/collagenⅠ/laminin/hepatic stellate cells分类
医药卫生引用本文复制引用
张玉峰,叶坤英,钟丹..小干扰RNA与反义寡核苷酸技术抑制肝星状细胞RhoA表达的效果比较[J].山东医药,2017,57(34):5-8,4.基金项目
河南省中医药科学研究专项课题(2014ZY02027). （2014ZY02027）
