反义TβRI对实验性大鼠肝纤维化的影响

Effects of antisense TβRI eukaryotic expressing plasmid on rat liver fibrosis

目的:观察反义转化生长因子βI型受体真核表达质粒对实验性大鼠肝纤维化的影响。方法:构建大鼠反义真核细胞表达质粒,导入大鼠肝纤维化模型体内,通过Ⅰ型胶原的免疫组化以及VG染色观察两种反义质粒联合作用对大鼠肝纤维化的影响。结果:反义TβRI表达质粒可减少受损肝脏中Ⅰ型胶原的沉积(P<0.05),并可使受损肝脏的病理形态有一定改善(P<0.05)。结论:反义TβRI真核表达质粒对肝纤维化的发展有一定的干预作用。

Objective： The pedonned experiments aimed to test the hypothesis that the introduction of anfisense transfonning growth factor β receptor Ⅰ （TβRI） eukaryotie expressing plasmids into a rat model of liver fibrosis may halt the progression of liver fibrosis.Methods： The fragments of TβRI cDNA was achieved by reverse transcription polymerase chain .ration （RT- PER）, and then amplified by nest PCR pcDNA3.1 （ ＋ ） - antisense TβRI eukaryotic expressing plasmid was construct by a directional inverted joins with the purified linear pcDNA3. 1 （ ＋ ） and the purified fragment of TβRI. Recombinant was identified by restriction endonuekease digestion and DNA sequence analysis. The recombinant plasmids were encapsulated by Lipofeetmine 2000, and then transduced into rats of liver fibrosis model via enterocoelia. The protein expression of type l collagen were evaluated by immunohistochemistry. VG staining was used for pathological diagnosis. Results： with the empty plasmid control group and disease control group, the deposition of collagens type Ⅰ decreased in anfisense TβRI group （436512.7 ± 130047.6, P 〈0.05; treatment control group 733917.7 ± 168290.8, disease control group 690236.1 ± 161333.7, normal control group 41329.7 ± 21321.9）. A significant different in the pathologic grade was found between normal control group and the other three groups （P〈 0.05）. A significant different in the pathologiegrade was found between disease control group and the antisense TβRI group（ P〈 0.05）. No difference was found in the pathologic grade between empty plasmid control group and disease control group （ P 〉 0.05）. Conclusion：Antisense TβRI eukaryotic expressing plasmid could inhibit the progress of liver fibrosis.