Chinese Medical E-ournals Database

Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition) ›› 2011, Vol. 07 ›› Issue (04): 303 -309. doi: 10.3877/cma.j.issn.1673-5250.2011.04.005

Original Article

Expression of Transforming Growth Factor-β1/Smad Signaling Pathway in Rat Glomerular Mesangial Cell and Interventional Effects of Angiotensin-Converting Enzyme Inhibitor

Ying DENG, Li YU, Rong-yan CHEN, Yue-qiang WEN, Jie WEN, Zhi-hong HAO   

  1. Department of Pediatrics, Guangzhou First Municipal People's Hospital, Guangzhou 510180, Guangdong Province, China
  • Published:2011-08-01
  • Supported by:
    * Project No. 2009-YB-028, supported by the Guangzhou Health Bureau Scientific Research Fund; project No. 5000424, supported by the Scientific and Technical Research Program for Medical Science of Guangdong Province
Objective

To investigate the role of transforming growth factor (TGF)-β1/Smad signaling pathway in glomerular mesangial cell (GMC), and the influence of angiotensin-converting enzyme inhibitor(ACEI) fosinopril (Fos) on the expression of phosphorylation-Smad2/3 (P-Smad2/3) and collagen Ⅰ(Col Ⅰ) in this way.

Methods

Glomerular mesangial cells of rat in vitro were cultured, and they were divided into 3 groups: ①TGF-β1 group (TGF-β1 5 ng/mL); ② Fos group (TGF-β1 5 ng/mL+ Fos 1×10-4 mol/L); ③control group (blank). Methyl thiazolyl tetrazolium (MTT) colorimetric method detected cellular proliferation. The change of phosphorylation-Smad2/3 expression was detected by indirect immunofluorescence. The expression of collagen Ⅰ in cell culture supernatant was detected by the enzyme-linked immunosorbent assay (ELISA).

Results

The optical density(OD) value of cellular proliferation was higher than that in TGF-β1 group when compared with control group(P<0.05), while lower in Fos group when compared with TGF-β1 group(P<0.05). Phosphorylation-Smad2/3 slightly expressed in control group, while increased obviously in TGF-β1 group compared with control group(P<0.05). In Fos group, the expression of phosphorylation-Smad2/3 were decreased apparently at 12 h and 24 h than those in TGF-β1 group(P<0.05).

Conclusion

TGF-β1 can activate the expression of phosphorylation-Smads and TGF-β1/Smad signaling pathway in glomerular mesangial cell, which will cause the increased expression of collagen Ⅰ and proliferation of glomerular mesangial cell. Fos can partially block the activation of transforming growth factor-β1/Smad signaling pathway in glomerular mesangial cell, inhibit the excretion of collagen Ⅰ and proliferation of glomerular mesangial cell.

图1 各组肾小球系膜细胞各时间点磷酸化-Smad2/3表达变化 (免疫荧光法,×200)
图2 各时间点ColⅠ蛋白表达量趋势图
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