Expression of hepatic stellate cells inhibited by receptor for advance glycoxidation end products specific small interfering RNA / 中华消化杂志

ABSTRACT

Objective To investigate the effect of specific small interfering RNA (siRNA) targeting receptor of advanced glycation end products (RAGE) on the production of fibrosis markers (laminin,hyaluronic acid (HA) and N-terminal procollagen Ⅲ propeptide (PⅢ NP) in hepatic fibrosis (HF).Methods The expression vectors of specific siRNA targeting RAGE were constructed.Primary rat hepatic stellar cells (HSC) were isolated and cultured.The primary rat HSC were transfected with the recombinant vector.The blank control group and unspecific siRNA vector pAKD-NC-transfected group were as controls.The expressions of RAGE,laminin,HA and PⅢ NP at mRNA and protein levels were detected by real-time polymerase chain reaction and Western blot,respectively.Least-significant difference (LSD) and Student-Newman-Keuls (SNK) were performed to analyze standard normal distribution or homogeneous variance.Non-normal distribution and heterogeneity of variance data were analyzed by non-parametric Wilcoxon test.Results The expressions of RAGE at mRNA and protein levels in pAKD-GR126-transfected primary HSC were (42.32 ± 6.16)％,(43.24±7.50)％,(51.06±13.79)％ and (47.94±5.36)％ in blank control group and pAKDNC group (F=7.791 and 36.513,all P＜0.05).The expressions of laminin at mRNA and protein levels were (41.07±3.13)％,(40.59±5.87)％,(53.89±2.25)％ and (52.46±4.68)％ in blank control group andpAKD-NC group (F=225.111 and 88.039,all P＜0.05).The expressions of HA at mRNA and protein levels were (45.69 ± 0.87) ％,(46.08 ± 2.36) ％,(54.20 ± 0.56) ％ and (52.30±3.42)％ in blank control group and pAKD-NC group (F=178.317 and 180.646,all P＜ 0.05).The expressions of PⅢ NP mRNA at mRNA and protein levels were (56.10±4.18)％,(55.15±2.39)％,(54.40±2.79)％ and (53.58±6.18)％ in blank control group and pAKD-NC group (F=141.633 and 49.670,all P＜0.05).Conclusion RAGE specific siRNA could inhibit the expression of RAGE in primary rat HSC and could significantly lower the expression of fibrosis markers laminin,HA and PⅢ NP at mRNA and protein level.