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Antisense oligonucleotides-based inhibition of gene and protein expression of tissue inhibitor of metalloproteinase-2 on experimental liver fibrosis in rats

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Author:: No author available

Journal Title:: WORLD CHINESE JOURNAL OF DIGESTOLOGY

Issue:: 23

DOI:: 10.3969/j.issn.1009-3079.2005.23.005

Key Word:: 反义寡核苷酸;金属蛋白酶组织抑制因子-2;肝纤维化;大鼠

Abstract：目的:探讨以TIMP-2为靶基因,应用反义寡核苷酸(asON)技术抑制其在肝组织中的表达对大鼠肝纤维化发展的影响.方法:22只大鼠分为治疗组(n=6)、模型组(n=6)和正常对照组(n=10).以人血白蛋白免疫攻击方法制备免疫诱导型肝纤维化大鼠模型.模型制备过程中,治疗组大鼠以尾静脉注射方式给予针对TIMP-2的硫代反义寡核苷酸.RT-PCR、原位杂交、免疫组化、ELISA等方法检测TIMP-2的转录以及表达水平.用病理学检查以及Ⅰ、Ⅳ型胶原的免疫组化结果分析肝纤维化发展程度.通过胶原纤维特殊染色及电镜等观察asON对大鼠肝纤维化的影响.结果:治疗组病理学分级状况优于模型组(u=2.071,P＜0.05).治疗组血清TIMP-2低于模型组(T=55,P＜0.05),高于正常对照组(T=55,P＜0.05).治疗组肝组织TIMP-2 mRNA表达弱于模型组(t=3.332,P＜0.05),高于正常对照组(t=5.550,P＜0.05).Ⅳ型胶原免疫组化图像定量分析结果显示,治疗组与模型组有显著差异(t=2.310,P＜0.05),其表达较低,但仍高于正常对照组(t=3.623,P＜0.05).治疗组Ⅰ型胶原免疫组化图像扫描结果与模型组相比,其数值亦较低(t=2.845,P＜0.05).治疗组肝窦基底膜胶原沉积较轻,与模型组无明显差异.结论:抑制TIMP-2在肝组织中的表达可以减缓大鼠肝纤维化发展.

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