Effect Of The Receptor For Advanced Glycation End Products-specific SiRNA On The Expressions Of Matrix Metalloproteinase-1 And Tissue Inhibitor Of Metalloproteinase-1

Objective The aim of the present study was to investigate the effect of the receptor for advanced glycation end products-specific siRNA on the expressions of matrix metalloproteinase-1(MMP-1) and tissue inhibitor of metalloproteinase-1(TIMP-1) in primary rat HSCs and HF rats.Method In vitro experiment, the primary rat HSCs were isolated and cultured. Then pAKD-GR126 which is small interfering RNA (siRNA) targeting RAGE was constructed and transfected to primary rat HSCs. We used the blank group and unspecific siRNA vector pAKD-NC-transfected group as controls. In vivo experiment, the liver fibrosis SD rats were induced by CCl4. Moreover, pAKD-GR126 was transfected to liver fibrosis SD rats at different doses via tail vein. The blank group, liver fibrosis model group and unspecific siRNA vector pAKD-NC-transfected group were used as controls. The Real-time quantitative PCR and Western-blot analyses were used to detect the expression levels of RAGE, MMP-1 and TIMP-1. The histological changes of rat livers were observed with H-E and Masson staining methods.Results In vitro experiment, the mRNA and protein expression levels of RAGE andTIMP-1 in the pAKD-GR126-transfected primary HSCs were significantly lower than those in the blank group and unspecific siRNA vector pAKD-NC-transfected groups(all P<0.05). But the mRNA and protein expression levels of MMP-1 in the pAKD-GR126-transfected primary HSCs was significantly higher than those in the blank group and pAKD-NC-transfected groups(all P<0.05). In vivo experiment, the mRNA and protein expression levels of RAGE and TIMP-1 in the low-, medium-, and high-does pAKD-GR126 treatment groups(LT, MT and HT) were more or less lower than those in the FM group(all P<0.05). But the mRNA and protein expression levels of MMP-1 were more or less higher than those in the FM group(all P<0.05). Compared with FM and NS groups, the fibrosis stage of rat livers became significantly lower, especially HT group.Conclusion The RAGE specific siRNA could effectively decrease the expression of RAGE and TIMP-1, but improve the expression of MMP-1 in primary rat HSCs and HF rats, which could reduce the degree of rat hepatic fibrosis.