[Changes of type I collagen and fibronectin expressions on cultured human Tenon's capsule fibroblasts transfected with Smad 7 vector].
OBJECTIVE
To investigate type I collagen and fibronectin expressions in the cultured human Tenon's capsular fibroblasts (HTFs) transfected with Smad 7 vector and to elucidate the possibility of Smad 7 in blocking tissue fibrosis after filtration surgery.


METHODS
Nucleofector(TM) was used to transfect Smad 7 vector into HTFs. The expressions of alpha2-type I procollagen (COL1A2) mRNA and fibronectin mRNA were detected by reverse transcription real-time quantitative polymerase chain reaction (real-time RT-PCR). The concentration of carboxyterminal propeptide of type I procollagen (PICP) in culture media was detected by radioimmunoassay. The above mentioned experiments were also detected before and after HTFs stimulated by TGF-beta(2) (10 ng/ml). HTFs and vector-HTFs were used as control groups.


RESULTS
Smad 7 was successfully transfected into HTFs evidenced by Smad 7 over expression. The expression of COL1A2 mRNA in Smad 7-HTFs was decreased 40.47% and 37.94%, respectively, when compared with the control HTFs and pCMV5-HTFs group. The PICP concentration of Smad 7-HTFs in the culture medium was decreased 52.10% and 57.35%, respectively, comparing to the two control groups. Furthermore, Smad 7-HTFs attenuated the increase of COL1A2 mRNA expression and PICP concentration of HTFs stimulated by TGF-beta(2). There was no difference in the fibronectin mRNA expressions between Smad 7-HTFs and the two control groups.


CONCLUSIONS
Smad 7 reduces the expression of COL1A2 mRNA and the synthesis of type I collagen, but has no effect on fibronectin mRNA expression, which may imply its ability to inhibit tissue fibrosis after filtration surgery.
Chen
Jun-yi
JY
Department of Ophthalmology, Eye & ENT Hospital of Fudan University, Shanghai 200031, China.
Sun
Xing-huai
XH
D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism*; D005123:Eye / Q000166:cytology; D005347:Fibroblasts / Q000378:metabolism*; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008297:Male; D008875:Middle Aged; D012333:RNA, Messenger / Q000235:genetics; D051906:Smad7 Protein / Q000235:genetics*; D014162:Transfection
D024042:Collagen Type I; D005353:Fibronectins; D012333:RNA, Messenger; C497806:SMAD7 protein, human; D051906:Smad7 Protein