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在无细胞体系和肝星状细胞中针对转化生长因子β1的核酶与U1小核核糖核酸嵌合核酶的活性鉴定

Activity identification of ribozyme and U1 snRNA chimeric ribozyme against TGFbeta1 in cell-free system and in hepatic stellate cells.

作者信息

Song Yuhu, Liu Fang, Tian Dean, Xue Xiulan, Liu Nanzhi, Wu Xiaoli, Lin Jusheng, Jin Youxin

机构信息

Institute of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.

出版信息

Sci China C Life Sci. 2006 Feb;49(1):73-81. doi: 10.1007/s11427-005-0036-8.

Abstract

Transforming growth factorbeta1 (TGFbeta1) is known to be intimately involved in many cellular processes. To explore the mechanism of TGFbeta1 in these processes, the non-chimeric hammerhead ribozyme and U1 snRNA chimeric ribozyme against TGFbeta1 were designed to down-regulate TGFbeta1 expression. The activity of non-chimeric ribozyme and U1 snRNA chimeric ribozyme against TGFbeta1 in vitro and in activated hepatic stellate cells (HSCs) was detected. Cleavage reactions of both ribozymes in vitro demonstrated that non-chimeric ribozyme possessed better cleavage activity in vitro than U1 snRNA chimeric ribozyme. The further study showed U1 snRNA chimeric ribozyme inhibited TGFbeta1 expression more efficiently than non-chimeric ribozyme in transfected HSC cells. So it indicates that the U1 snRNA chimeric ribozyme provides an alternative approach for the research on the precise mechanism of TGFbeta1 in many cellular processes and a potential therapeutic candidate for TGFbeta1-related diseases.

摘要

已知转化生长因子β1(TGFβ1)密切参与许多细胞过程。为了探究TGFβ1在这些过程中的作用机制,设计了针对TGFβ1的非嵌合锤头状核酶和U1 snRNA嵌合核酶,以下调TGFβ1的表达。检测了非嵌合核酶和U1 snRNA嵌合核酶在体外及活化肝星状细胞(HSCs)中针对TGFβ1的活性。两种核酶在体外的切割反应表明,非嵌合核酶在体外具有比U1 snRNA嵌合核酶更好的切割活性。进一步研究表明,在转染的HSC细胞中,U1 snRNA嵌合核酶比非嵌合核酶更有效地抑制TGFβ1的表达。因此,这表明U1 snRNA嵌合核酶为研究TGFβ1在许多细胞过程中的精确机制提供了一种替代方法,并且是TGFβ1相关疾病的潜在治疗候选物。

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