Abstract
In this study, influence caused by expression plasmids of connective tissue growth factor (CTGF) and tissue inhibitor of metalloproteinase-1 (TIMP-1) short hairpin RNA (shRNA) on mRNA expression of CTGF,TIMP-1,procol-α1 and PCIII in hepatic tissue with hepatic fibrosis, a precancerous condition, in rats is analyzed. To screen and construct shRNA expression plasimid which effectively interferes RNA targets of CTGF and TIMP-1 in rats. 50 cleaning Wistar male rats are allocated randomly at 5 different groups after precancerous fibrosis models and then injection of shRNA expression plasimids. Plasmid psiRNA-GFP-Com (CTGF and TIMP-1 included), psiRNA-GFP-CTGF, psiRNA-GFP-TIMP-1 and psiRNA- DUO-GFPzeo of blank plasmid are injected at group A, B, C and D, respectively, and as model control group that none plasimid is injected at group E. In 2 weeks after last injection, to hepatic tissue at different groups, protein expression of CTGF, TIMP-1, procol-α1and PC III is tested by immunohistochemical method and,mRNA expression of CTGF,TIMP-1,procol-α1 and PCIII is measured by real-time PCR. One-way ANOVA is used to comparison between-groups. Compared with model group, there is no obvious difference of mRNA expression among CTGF,TIMP-1,procol-α1,PC III and of protein expression among CTGF, TIMP-1, procol-α1, PC III in hepatic tissue at group injected with blank plasmid. Expression quantity of mRNA of CTGF, TIMP-1, procol-α1 and PCIII at group A, B and C decreases, protein expression of CTGF, TIMP-1, procol-α1, PC III in hepatic tissue is lower, where the inhibition of combination RNA interference group (group A) on procol-α1 mRNA transcription and procol-α1 protein expression is superior to that of single interference group (group B and C) (P<0.01 or P<0.05). RNA interference on CTGF and/or TIMP-1 is obviously a inhibiting factor for mRNA and protein expression of CTGF, TIMP-1, procol-α1 and PCIII. Combination RNA interference on genes of CTGF and TIMP-1 is superior to that of single RNA interference, and this could be a contribution for prevention of precancerous condition.
Highlights
Many past studies suggested that transforming growth factor β (TGF-β1) is the strongest factor prompting hepatic fibrosis in the activation of hepatic stellate cell (HSC) that other cell-factors participated in, block of signal pathway of TGF-β1 is considered to be the best option for hepatic fibrosis therapy (Lee et al, 2011)
In 2 weeks after last injection, to hepatic tissue at different groups, protein expression of connective tissue growth factor (CTGF), tissue inhibitor of metalloproteinase-1 (TIMP-1), procol-α1and PC III is tested by immunohistochemical method and,mRNA expression of CTGF,TIMP-1,procol-α1 and PCIII is measured by real-time PCR
DMN is chartered by hepatic genotoxicity, the hepatic model induced by DMN has advantages such as short modeling time, lower mortality, stable formation of hepatic fibrosis, less selfhealing trend without induction, and similar situation to earlier human hepatic fibrosis etc (Kuang et al, 2008)
Summary
Many past studies suggested that transforming growth factor β (TGF-β1) is the strongest factor prompting hepatic fibrosis in the activation of hepatic stellate cell (HSC) that other cell-factors participated in, block of signal pathway of TGF-β1 is considered to be the best option for hepatic fibrosis therapy (Lee et al, 2011). This study uses recombinant plasmids constructed previously with CTGFshRNA and TIMP-1shRNA respectively of psiRNA-GFP-CTGF, psiRNA- GFPTIMP-1 and psiRNA- GFP-Com (CTGF and TIMP1included), to transfect rats with hepatic fibrosis. Further analyzes their influence on mRNA and proteinrelated expression of CTGF and TIMP-1, to prepare to study the roles of CTGF and TIMP-1 of TGF-β1 downstream effect medium in hepatic fibrosis and explore the gene therapy for hepatic fibrosis. Influence caused by expression plasmids of connective tissue growth factor (CTGF) and tissue inhibitor of metalloproteinase-1 (TIMP-1) short hairpin RNA (shRNA) on mRNA expression of CTGF,TIMP-1,procol-α1 and PCIII in hepatic tissue with hepatic fibrosis, a precancerous condition, in rats is analyzed. Combination RNA interference on genes of CTGF and TIMP-1 is superior to that of single RNA interference, and this could be a contribution for prevention of precancerous condition
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callMore From: Asian Pacific journal of cancer prevention : APJCP
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
