[Effects of antisense transforming growth factor beta receptor-I expressing plasmid on pig serum-induced rat liver fibrosis].
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.
Jiang
Wei
W
Department of Gastroenterology, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
Wang
Jiyao
J
Yang
Changqing
C
Wang
Yiqing
Y
Liu
Wenbin
W
He
Boming
B
D030201:Activin Receptors, Type I / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D001769:Blood / Q000276:immunology; D024042:Collagen Type I / Q000378:metabolism; D024061:Collagen Type III / Q000378:metabolism; D016373:DNA, Antisense / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods; D008103:Liver Cirrhosis / Q000235:genetics / Q000276:immunology / Q000628:therapy*; D008297:Male; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D017346:Protein Serine-Threonine Kinases; D051381:Rats; D017207:Rats, Sprague-Dawley; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta / Q000235:genetics* / Q000378:metabolism; D013552:Swine; D016212:Transforming Growth Factor beta / Q000097:blood; D053773:Transforming Growth Factor beta1
D024042:Collagen Type I; D024061:Collagen Type III; D016373:DNA, Antisense; D018125:Receptors, Transforming Growth Factor beta; C508962:Tgfb1 protein, rat; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D017346:Protein Serine-Threonine Kinases; D030201:Activin Receptors, Type I; D000077293:Receptor, Transforming Growth Factor-beta Type I; C000625843:Tgfbr1 protein, rat