摘要
目的研究靶向小干扰RNA沉默转化生长因子β1（TGFβ1）对肝纤维化大鼠体内血小板衍生生长因子D（PDGF-BB）及其受体（PDGF-DR）表达的影响。方法将40只雄性SD大鼠随机分为4组：正常对照组、模型组、TGEp1siRNA治疗组、阴性对照组。除正常对照组外，其余各组均给予40%四氯化碳皮下注射诱导大鼠肝纤维化模型。阴性对照组和TGFp1siRNA治疗组经大鼠尾静脉分别按0．25mg／kg注射阴性对照质粒、TGFD1siRNA质粒，正常组与模型组注射无菌等渗盐水，每周2次。12周后处死大鼠，取肝组织样本，分别采用Real—TimePCR、免疫组织化学及Westernblot检测PDGF—BB、PDGF—βR及磷酸化细胞外调节蛋白激酶（p-ERK1／2）在大鼠肝组织的表达。据资料不同分别采用单因素方差分析、口检验或Kmskal—Wallis检验进行统计学分析。结果与模型组、阴性对照组相比，TGED1siRNA能显著抑制肝组织中PDGF—βB及PDGF-DR的mRNA和蛋白的表达（F值分别为24．785、22．920，P〈0．01）；TGFp1siRNA还能明显抑制肝组织中p-ERK1／2的表达（P〈0．05）。结论靶向TGFD1siRNA可以有效地下调肝组织中PDGF—BB、PDGF—βR及p-ERK1／2的表达从而起到改善肝纤维化的作用。
Objective To investigate the impact of transforming growth factor-β1 （TGF-β1） silencing by small interfering RNA （siRNA） on the expression of platelet-derived growth factor-β （PDGF-βB） and its receptor （PDGF-βR） in rats with hepatic fibrosis. Methods A total of 40 male Sprague-Dawley rats were randomly divided into normal control group, model group, TGF-β1 siRNA treatment group, and negative control group. All rats except those in the normal control group were given subcutaneous injection of 40% carbon tetrachloride to establish a rat model of hepatic fibrosis. The rats in the negative control group and theTGF-β1 siRNA treatment group were given tail vein injection of negative control plasmid or TGF-β1 siRNA plasmid twice a week at a dose of 0.25 mg/kg, and those in the normal control group and the model group were given the injection of sterile isotonic saline twice a week. The rats were sacrificed after 12 weeks and liver tissue samples were collected. Real-time PCR, immunohistochemistry, and Western blot were used to measure the expression of PDGF-BB, PDGF-βR, and phosphorylated extracellular signal-regulated kinase 1/2 （p-ERK1/2） in liver tissue. A one-way analysis of variance, the q test, and the Kruskal-Wallis test were used for statistical analysis based on data type. Results Compared with the model group and the negative control group, the TGF-β1 siRNA treatment group had significantly inhibited mRNA and protein expression of PDGF-BB and PDGF-βR （F = 24.785 and 22.92, P 〈 0.01）, as well as significantly inhibited expression of p-ERK1/2 （P 〈 0.05）. Conclusion Targeted TGF-β1 siRNA can effectively downregulate the expression of PDGF-BB, PDGF-βR, and p-ERK1/2 in liver tissue and thus help to improve hepatic fibrosis.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2017年第9期701-705,共5页
Chinese Journal of Hepatology
基金
国家自然科学基金（81270503）
关键词
肝硬化
转化生长因子Β1
血小板衍生生长因子
细胞外调节蛋白激酶
Hepatic cirrhosis
Transforming growth factor-beta 1
Platelet-derived growth factor
Extracellular regulated protein kinase
