[Changes of fibronectin and type IV collagen expression in cultured rat mesangial cells transfected with Smad 2 vector].
OBJECTIVE
To study the changes of fibronectin (FN) and type IV collagen (ColIV) expression in cultured rat mesangial cells (MsC) transfected with Smad 2 vector and to investigate the molecular mechanism of glomerular extracellular matrix accumulation in glomerulosclerosis via transforming growth factor-beta (TGF-beta)/Smad signal pathway.


METHODS
Smad 2 vector was transfected into MsC by calcium phosphate. Western blot analysis was used to detect Smad 2 protein. The expression of FN and ColIV proteins and their mRNAs was determined by Western blot and reverse transcriptase-polymerase chain reaction respectively.


RESULTS
Four MsC clones (T-12, T-31, T-35, T-40) with Smad 2 overexpression were established. The expression of FN and ColIV was significantly increased at mRNA and protein levels in two (T-12, T-31). Compared with controls, the expression of FN proteins and mRNAs in these two clones was 2.4 times (P < 0.05) and 2.7 times (P < 0.05) higher respectively. The expression of ColIV proteins and mRNAs was 2.9 times (P < 0.01) and 3.3 times (P < 0.01) higher respectively.


CONCLUSIONS
It is postulated that Smad 2 in TGF-beta/Smad signal pathway is important in promoting the accumulation of FN and ColIV in sclerotic glomeruli of diseased kidneys.
Wang
Xiao-Gang
XG
Department of Pathology, Shanghai Medical College, Fudan University, Shanghai 200032, China.
Yu
Hong
H
Chen
Qi
Q
Zhao
Zhong-Hua
ZH
Zhang
Zhi-Gang
ZG
Guo
Mu-Yi
MY
D000818:Animals; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000096:biosynthesis* / Q000235:genetics; D005353:Fibronectins / Q000096:biosynthesis* / Q000235:genetics; D005822:Genetic Vectors; D050527:Mesangial Cells / Q000166:cytology / Q000378:metabolism*; D010957:Plasmids; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D015398:Signal Transduction; D051899:Smad2 Protein / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000378:metabolism
D024141:Collagen Type IV; D005353:Fibronectins; D012333:RNA, Messenger; D051899:Smad2 Protein; D016212:Transforming Growth Factor beta