[Changes of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 expressions on cultured rat mesangial cells transfected with Smad 7 vector].
OBJECTIVE
To investigate matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) expressions in the cultured rat mesangial cells (MsC) transfected with Smad 7 vector and to elucidate the mechanism of Smad 7 in blocking tissue fibrosis.


METHODS
Lipofectin method was used to transfect Smad 7 vector into MsC. Western blot and RT-PCR analyses were then used to detect Smad 7 protein and mRNA expression levels. The expressions of MMP-2 and TIMP-2 were determined by Western blot, RT-PCR and zymography assay.


RESULTS
Two MsC clones (S-22, S-26) with Smad 7 overexpression were successfully established. The two clones showed an increased expression of MMP-2 protein and enhanced enzyme activity. The expressions of TIMP-2 protein and mRNA however were suppressed.


CONCLUSIONS
It is possible that Smad 7 can alleviate the development of tissue fibrosis by upregulating the expression of MMP-2 and downregulating the expression of TIMP-2 in mesangial cells.
Yu
Hong
H
Department of Pathology, Shanghai Medical College, Fudan University, Shanghai 200032, China.
Wang
Xiao-gang
XG
Wang
Yi
Y
Chen
Qi
Q
Zhang
Xiu-rong
XR
Guo
Mu-yi
MY
D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D005920:Glomerular Mesangium / Q000166:cytology / Q000378:metabolism*; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051906:Smad7 Protein; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics / Q000378:metabolism*; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*
D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D051906:Smad7 Protein; C497808:Smad7 protein, rat; D015534:Trans-Activators; D019716:Tissue Inhibitor of Metalloproteinase-2; D020778:Matrix Metalloproteinase 2