Effects of centellaasiatica granule on the expression of Smad 2/3, Smad 7 and collagen Ⅳ in the mesangial cells stably expressed TGF-β1.
10.12047/j.cjap.5568.2018.030
- Author:
Ji-Wei MA
1
;
Hong-Tian WANG
1
;
Hao-Fei LIU
1
;
Lei-Peng DONG
1
;
Yuan DING
2
;
Ji-Qiong BAI
3
;
Zhu ZHANG
1
;
Li-Jie DONG
4
Author Information
1. Department of Nephrology, Henan University of Traditional Chinese Medicine Affiliated First Hospital, Zhengzhou 450000.
2. Xinyang Central Hospital, Xinyang 464000.
3. Gongyi City People's Hospital, Gongyi 451200.
4. Tianjin Medical University Eye Hospital, Eye Institute, Tianjin 300384, China.
- Publication Type:Journal Article
- Keywords:
Smad signaling pathway;
centellaasiatica;
collagen Ⅳ;
diabetic nephropathy;
rat;
transforming growth factor-beta 1
- MeSH:
Animals;
Cells, Cultured;
Centella;
chemistry;
Collagen Type IV;
metabolism;
Drugs, Chinese Herbal;
pharmacology;
Mesangial Cells;
drug effects;
metabolism;
Rats;
Signal Transduction;
Smad Proteins;
metabolism;
Smad2 Protein;
metabolism;
Smad3 Protein;
metabolism;
Smad7 Protein;
metabolism;
Transforming Growth Factor beta1;
metabolism
- From:
Chinese Journal of Applied Physiology
2018;34(2):122-125
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVES:Stably expressed transforming growth factor -beta 1(TGF-β1)MCs were obtained and the effects of centellaasiatica (CA) granule on the expressions of Smad 2/3, Smad 7 and collagen Ⅳ and the level of Smad 2/3 phosphorylation were observed.
METHODS:Lipofectin method was used to transfect TGF-β1 vector into MC, and the stably expressed TGF-β1 cell lines were selected by G418. The cells were divided into three groups. Control group:normal MC + RPMI 1640 + 10% normal rat serum; TGF-β1 group:stably expressed TGF-β1 MC + RPMI 1640 + 10% normal rat serum; CA group:stably expressed TGF-β1 MC + RPMI 1640 + 10% rat serum containing high CA. The experiments were repeated for five times. The contents of TGF-β1 and collagen Ⅳ in the culture medium were detected with ELISA, the expressions of mRNA and protein of TGF-β1, Smad 2/3, Smad 7 and the level of Smad 2/3 phosphorylation were detected by using real time quantitative polymerase chain reaction and Western blot.
RESULTS:The contents of TGF-β1 and collagen Ⅳ in the culture medium of stably-expressed TGF-β1 MC were increased significantly, and the CA could reverse the effects of TGF-β1. The expressions of mRNA and protein of TGF-β1, Smad 2/3 and the level of Smad 2/3 phosphorylation were increased significantly in TGF-β1 transfected MC, and CA could dramatically reduce the expressions of mRNA and protein of TGF-β1, Smad 2/3 and the level of Smad 2/3 phosphorylation. The high expression of TGF-β1 decreased the expression of Smad 7 mRNA and protein, and the CA could antagonize the effect of mRNA expression.
CONCLUSIONS:The MCs stably-expressed TGF-β1 can activate the TGF-β1/Smad signal pathway and increase the expression of collagen Ⅳ. CA can decrease the occurrence of diabetic nephropathy(DN) by reducing the production of collagen Ⅳ through inhibiting the TGF-β1/Smad signal pathway.
