JOURNAL BROWSE
Search
Advanced SearchSearch Tips
Influence of Expression Plasmid of Connective Tissue Growth Factor and Tissue Inhibitor of Metalloproteinase-1 shRNA on Hepatic Precancerous Fibrosis in Rats
facebook(new window)  Pirnt(new window) E-mail(new window) Excel Download
 Title & Authors
Influence of Expression Plasmid of Connective Tissue Growth Factor and Tissue Inhibitor of Metalloproteinase-1 shRNA on Hepatic Precancerous Fibrosis in Rats
Zhang, Qun; Shu, Fu-li; Jiang, Yu-Feng; Huang, Xin-En;
  PDF(new window)
 Abstract
Background: 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- and PCIII in hepatic tissue with hepatic fibrosis, a precancerous condition, in rats is analyzed. Materials and Methods: 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- and PC III is tested by immunohistochemical method and,mRNA expression of CTGF,TIMP-1,procol- and PCIII is measured by real-time PCR. One-way ANOVA is used to comparison between-groups. Results: Compared with model group, there is no obvious difference of mRNA expression among CTGF,TIMP-1,procol-, PC III and of protein expression among CTGF, TIMP-1, procol-, PC III in hepatic tissue at group injected with blank plasmid. Expression quantity of mRNA of CTGF, TIMP-1, procol- and PCIII at group A, B and C decreases, protein expression of CTGF, TIMP-1, procol-, PC III in hepatic tissue is lower, where the inhibition of combination RNA interference group (group A) on procol- mRNA transcription and procol- protein expression is superior to that of single interference group (group B and C) (P<0.01 or P<0.05). Conclusions: RNA interference on CTGF and/or TIMP-1 is obviously a inhibiting factor for mRNA and protein expression of CTGF, TIMP-1, procol- 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.
 Keywords
RNA interference;connective tissue growth factor;tissue inhibitor of metalloproteinase-1;
 Language
English
 Cited by
 References
1.
Giannelli G, Mazzocca A, Fransvea E, et al (2011). Inhibiting TGF-${\beta}$1 signaling in hepatocelluar carcnoma. Biochim Biophys Acta, 1815, 214-23.

2.
Hepatic fibrosis group of chinese society of hepatology (2002). consensus about evaluating hepatic fibrosis diagnosis and efficacy. Chinese J Hepatol, 10, 327-328.

3.
Hemmann S, Graf J, Roderfeld M, et al (2007). Expression of MMPs and TIMPs in liver fibrosis-asystematic review with special emphasis on anti-fibrotic strategies. J Hepatol, 46, 955-75. crossref(new window)

4.
Jiang YF, Huang F, Zhang JJ, et al (2011). RNA interference targets on connective tissue growth factor and tissue inhibitor tissue inhibtor of metalloproteinase-1 in vitro. J Clin Hepatol, 14, 246-8.

5.
Jiang YF, Liu JQ, Zhang JJ, et al (2011). Construction of two different hepatic fibrosis-related cytokines shRNA recombinant plasmid. Chongqing Med, 40, 2341-2.

6.
Jiang YF, Zhou DJ, Zhang JJ, et al (2012). Construction of dual RNA interference plasmid of hepatic fibrosis-related cytokines and study on transfection of hepatic stellate cells. J Clin Hepatol, 15, 516-9.

7.
Kuang SL, Hu B (2008). Development of study on model of hepatic fibrosis in rats. Laboratory Animal Comparative Med, 28, 62-66.

8.
Lee UE, Friedman SL (2011). Mechanisms of hepatic fibrogenisis. Best Pract Res Clin Gastroenterol, 25, 195-206. crossref(new window)

9.
Rettiq GR,Behlke MA (2012). Progress toward in vivo use of siRNA-II. Mol Ther, 20, 483-512. crossref(new window)

10.
Tache D,Boqdan F,Pisoschi C, et al (2011). Evidence for the involvement of TGF-${\beta}$1-CTGF axis in liver fibrogenisis secondary to hepatic viral infection. Rom J Morphol Embryol, 52, 409-12.

11.
Xu LM (2010). Update on hepatic fibrosis research. J Clin Hepatol, 18, 563-565.

12.
Yang J,Zheng J,Wu L, et al (2011). MDRG2 ameliorates hepatic fibrosis by inhibiting the TGF-${\beta}$1/Smad pathway and altering the MMP2/TIMP2 ratio in rats. PLoS One,6, 27710. crossref(new window)

13.
Yan X,Chen YG (2011). Smad7:not only a regulator,but also a cross-talk mediator of TGF-${\beta}$ signaling. Biochem J, 434, 1-10. crossref(new window)