引用本文:张 敏,江宇杰,石小枫,刘 杞,孙 航.基质金属蛋白酶抑制剂1和2 siRNA对CCl4诱导的肝纤维化大鼠Smad7蛋白表达的影响[J].重庆医科大学学报,2014,38(7):919~922
基质金属蛋白酶抑制剂1和2 siRNA对CCl4诱导的肝纤维化大鼠Smad7蛋白表达的影响
Effect of tissue inhibitor of metalloproteinases 1 and 2 siRNA on the expression of Smad7 protein in hepatic fibrosis rats
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中文关键词:  siRNA  肝纤维化  Smad7  基质金属蛋白酶抑制剂
英文关键词:siRNA  liver fibrosis  Smad7  tissue inhibitor of metalloproteinases
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张 敏,江宇杰,石小枫,刘 杞,孙 航 重庆医科大学附属第二医院感染科、教育部感染性疾病分子生物学重点实验室重庆 400010 
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中文摘要:
      目的:研究基质金属蛋白酶抑制剂(tissue inhibitor of metalloproteinases,TIMP)1和2的siRNA对CCl4诱导的肝纤维化大鼠Smad7蛋白表达的影响。方法:采用前期构建的TIMP-1 siRNA、TIMP-2 siRNA治疗肝纤维化大鼠的肝脏组织作为研究对象,分为正常组、模型组、阴性对照组(0.25 mg/kg非特异性靶序列)、TIMP-1 siRNA 0. 25 mg/kg组和TIMP-2 siRNA 0.25 mg/kg组。Real-time PCR法检测Smad7的mRNA表达;免疫组织化学、Western blot检测Smad7的蛋白表达。结果:Real-time PCR结果显示TIMP-1 siRNA 0.25 mg/kg组(59.7±3.3)%、TIMP-2 siRNA 0.25 mg/kg组(60.2±3.4)% Smad7的mRNA表达量均较模型组(28.0±1.6)%、阴性对照组(28.6±1.7)%明显增加(P=0.000);免疫组织化学结果显示TIMP-1 siRNA 0.25mg/kg组(8.55±0.67)、TIMP-2 siRNA 0.25 mg/kg组(8.23±0.85)Smad7的蛋白表达量均较模型组(4.25±0.53)、阴性对照组(4.19±0.55)明显增加(P=0.000)。Western blot结果显示TIMP-1 siRNA 0.25 mg/kg组(0.706±0.039)、TIMP-2 siRNA 0.25 mg/kg组(0.698±0.038)Smad7的蛋白表达量均较模型组(0.278±0.013)、阴性对照组(0.285±0.014)明显增加(P=0.000)。结论:TIMP-1 siRNA、TIMP-2 siRNA对于肝纤维化Smad7的表达有促进作用。
英文摘要:
      Objective:To observe the expression of Smad7 protein in carbon tetrachloride(CCl4)-induced hepatic fibrosis rats treated with TIMP-1 short interference RNA(siRNA) and TIMP-2 siRNA. Methods:The hepatic tissues collected from hepatic fibrosis rats were divided into 5 groups:normal group,model group,negative control group,TIMP-1 siRNA 0.250 mg/kg group and TIMP-2 siRNA 0.250 mg/kg group. The expression of Smad7 was detected by real-time PCR,immunohistochemistry and Western blot. Results:Real-time PCR analysis showed that mRNA expression of Smad7 was significantly higher in TIMP-1 siRNA 0.250 mg/kg group(59.7±3.3)% and TIMP-2 siRNA 0.250 mg/kg group(60.2±3.4)% than in model group(28.0±1.6)% and negative control group(28.6±1.7)%(P=0.000). Immunohistochemistry showed that protein expression of Smad7 was significantly higher in TIMP-1 siRNA 0.250 mg/kg group(8.55±0.67) and TIMP-2 siRNA 0.250 mg/kg group(8.23±0.85) than in model group(4.25±0.53) and negative control group(4.19±0.55)(P=0.000). Western blot showed that protein expression of Smad7 was significantly higher in TIMP-1 siRNA 0.250 mg/kg group(0.706±0.039) and TIMP-2 siRNA 0.250 mg/kg group(0.698±0.038) than in the model group(0.278±0.013) and negative control group(0.285±0.014)(P=0.000). Conclusion:Silencing the TIMP-1 and TIMP-2 with siRNA could be involved in the improvement of the hepatic fibrosis in rats.
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