Chinese herb Cordyceps polysaccharide inhibits transforming growth factor β1-induced epithelial-to-mesenchymal transition in proximal tubular epithelial cells
- VernacularTitle:虫草多糖对转化生长因子β1诱导人近端肾小管上皮细胞间充质转分化的影响
- Author:
Zhuang GONG
;
Xiaoliang ZHANG
;
Jun GAO
;
Qing LI
;
Bicheng LIU
- Publication Type:Journal Article
- Keywords:
Fibrosis;
Transforming growth factor β;
Epithelial cells;
Kidneytubules,proximal;
Cordyceps polysaccharide;
HK-2 cell;
Epithelial-to-mesenchymaltransition
- From:
Chinese Journal of Nephrology
2008;24(9):651-656
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the influence of Cordyceps polysaceharide (Cp) on epithelial-to-mesenchymal transition (EMT) induced by transforming growth factor-β1 (TGF-β1)in proximal tubular epithelial cells (PTEC). Methods HK-2 cell proliferation was determined by MTT assay. After incubation of HK2 cells with increasing concentrations of TGF-β1 (0, 0.1, 0.5, 1, 5, 10 μg/L) at 48 h and with TGF-β1 (5 μ/L) at different time points, E-cadherin, α-SMA, FN expression at transcriptional and protein levels were detected by real-time PCR and Western blotting respectively. The cells were pretreated with 1,5, 10 g/L Cp respectively for 24 h before adding TGF-β1 (5μg/L), then the cells were incubated for additional 48 h, mRNA and proteinexpression of above 3 cytokines was examined by real-time PCR and Western blotting as well. Results CP alone (0.01, 0.1, 1,5, 10 g/L) induced HK-2 cell proliferation in a dose-dependent manner. TGF-β1 enhanced α-SMA, FN expression while inhibited E-cedherin expression at both transcriptional and" protein level in HK-2 cell. At transcriptional level, compared to single TGF-β1 (5 μg/L) stimulating group, after Cp (1,5, 10 g/L) pretreatment for 24 h, the inhibition rate of a-SMA mRNA was 37.98%, 68.08% and 84.36%, respectively; FN mRNA was 46.97%, 63.82% and 81.85%, respectively; E-cadherin was up-regulated by 0.67 fold, 2.69 folds and 5.43 folds, respectively (P<0.05). At protein level, the inhibition rate of α-SMA was 33.40%, 47.75% and 68.50%, respectively; FN was 16.26%, 65.92% and 80.30%, respectively; E-cadherin was up-regulated by 1.33 folds, 3.19 folds and 4.29 folds, respectively (all P<0.05). Under Light microscopy, the Cp reversed cell shape from spindle-shape induced by TGF-β1 to nearly normal shape. Conclusion Cp may exert its inhibitive effects on TGF-β1-induced EMT.