Effect of ribozyme against transforming growth factorbeta1 on biological character of activated HSCs.
Transforming growth factorbeta1 (TGFbeta1) is considered to be the principal contributor to liver fibrosis. So in this study the ribozymes against TGFbeta1 were designed. The in vitro cleavage activities of the ribozymes were assayed through incubation of (32)p-labeled target RNAs and (32)p-labeled ribozymes in different conditions. HSC-T6 cells were transfected with the eukaryotic constructs encoding ribozyme and disable ribozyme, then the stable cell clones were used to evaluate its antifibrotic characteristic through the effect of ribozyme on biological character of activated hepatic stellate cells (HSCs). The results demonstrated that two ribozymes (Rz803 and Rz1395) could cleave target RNAs into expected products effectively, Rz803 possessed better cleavage activity in vitro. Stable transfection of Rz803 into activated HSCs reduced TGFbeta1 expression in mRNA and protein level efficiently. The further studies demonstrated that Rz803 reduced deposition of collagen I, suppressed HSC proliferation, but had no effect on HSC activation in transfected HSC-T6 cells. Therefore, it indicated that Rz803 could reverse the character of activated HSCs by down-regulating TGFbeta1 expression efficiently and diminishing TGFbeta1 signaling underlying activation of hepatic stellate cells. As the consequence, it would provide a potential therapeutic approach for liver fibrosis.
Song
Yu Hu
YH
Institute of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China.
Zhou
Xiu Ming
XM
Xue
Xiu Nan
XN
Liu
Nan Zhi
NZ
Tian
De An
DA
Kong
Xin Juan
XJ
Wu
Xiao Li
XL
Lin
Ju Sheng
JS
Jin
You Xin
YX
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D017931:DNA Primers; D005822:Genetic Vectors; D022781:Hepatocytes / Q000187:drug effects*; D007700:Kinetics; D008103:Liver Cirrhosis / Q000378:metabolism*; D012313:RNA / Q000187:drug effects / Q000378:metabolism*; D016337:RNA, Catalytic / Q000494:pharmacology*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects*; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000378:metabolism*
D017931:DNA Primers; D016337:RNA, Catalytic; D016212:Transforming Growth Factor beta; C427204:hammerhead ribozyme; D012313:RNA