[反义转化生长因子β受体-I表达质粒对猪血清诱导的大鼠肝纤维化的影响]

[Effects of antisense transforming growth factor beta receptor-I expressing plasmid on pig serum-induced rat liver fibrosis].

作者信息

Jiang Wei, Wang Jiyao, Yang Changqing, Wang Yiqing, Liu Wenbin, He Boming

机构信息

Department of Gastroenterology, Zhongshan Hospital, Fudan University, Shanghai 200032, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2002 Sep 10;82(17):1160-4.

PMID:12475400

Abstract

OBJECTIVE

To observe the effects of antisense transforming growth factor beta receptor-I (TbetaRI) expressing plasmid on rat liver fibrosis.

METHODS

RT-nested-PCR and gene recombinant techniques were used to construct the rat antisense TbetaRI recombinant plasmid which can be expressed in eucaryotic cells. The recombinant plasmid and blanx vector (pcDNA3) were encapsulated by glycosyl-poly-L-lysine and then transducted into rats of pig serum-induced liver fibrosis model respectively. Expression of exogenous transfected plasmid was assessed by Northern blot, RT-PCR, Western blot and immunohistochemistry. Serum TGF-beta(1) was determined by ELISA. The content of hepatic hydroxyproline was tested. Immunohistochemistry was used to test type I and III collagen. VG staining was used for pathological study.

RESULTS

The exogenous antisense TbetaRI could be expressed in vivo, and could block the mRNA and protein expression of TbetaRI in the fibrotic liver induced by pig serum. Its expression also reduced the level of TGF-beta(1) (P < 0.05) and decreased the the contents of hepatic hydroxyproline and the deposition of collagens type I and type III (P < 0.01). And its expression also improved the pathologic classification of liver fibrosis models (P < 0.01).

CONCLUSIONS

The results demonstrate that the antisense TbetaRI recombinant plasmid has certain extern reverse effect on liver fibrosis and make it as a possible candidate for use in future gene therapy.

摘要

目的

观察反义转化生长因子β受体I（TβRI）表达质粒对大鼠肝纤维化的影响。

方法

采用RT - 巢式PCR和基因重组技术构建可在真核细胞中表达的大鼠反义TβRI重组质粒。将重组质粒和空白载体（pcDNA3）用糖基化聚 - L - 赖氨酸包裹，然后分别转导入猪血清诱导的肝纤维化模型大鼠体内。通过Northern印迹、RT - PCR、Western印迹和免疫组织化学评估外源转染质粒的表达。用ELISA法测定血清TGF -β(1)。检测肝组织羟脯氨酸含量。用免疫组织化学检测I型和III型胶原。用VG染色进行病理研究。