Changes of fibronectin and type IV collagen expression in cultured rat mesangial cells transfected with Smad 2 vector.
- Author:
Xiao-Gang WANG
1
;
Hong YU
;
Qi CHEN
;
Zhong-Hua ZHAO
;
Zhi-Gang ZHANG
;
Mu-Yi GUO
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cells, Cultured; Collagen Type IV; biosynthesis; genetics; Fibronectins; biosynthesis; genetics; Genetic Vectors; Mesangial Cells; cytology; metabolism; Plasmids; RNA, Messenger; biosynthesis; genetics; Rats; Signal Transduction; Smad2 Protein; genetics; metabolism; Transfection; Transforming Growth Factor beta; metabolism
- From: Chinese Journal of Pathology 2005;34(2):97-100
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSSmad 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.
RESULTSFour 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.
CONCLUSIONSIt 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.
