Original articleTGF-β1 and integrin synergistically facilitate the differentiation of rat podocytes by increasing α-smooth muscle actin expression
Section snippets
Podocyte cell culture
To generate primary cultures of rat podocytes, glomeruli were isolated from Sprague–Dawley rats (50-g body weight) by the sieving method.31, 32 Renal cortical tissue homogenates were carefully passed through sequential stainless-steel sieves of 212 μm and then 106-μm pore size. The isolated glomeruli were collected from a sieve of 75-μm pore size and inspected with phase contrast microscopy for confirmation of successful extraction. Isolated glomeruli were plated in plastic culture flasks that
α-SMA protein expression of podocytes after TGF-β1 stimulation
The expression of α-SMA protein induced by TGF-β1 stimulation for 4 days was analyzed by Western blotting. Western blotting of cell lysates showed that TGF-β1 induced a dose-dependent increase in the expression of α-SMA protein in cultured rat podocytes. A faint band existed for α-SMA protein in podocytes grown on collagen-coated plates with free-serum medium. A significant increase of α-SMA protein occurred after TGF-β1 stimulation with dosage of 2.5, 7.5, 10, 25, and 50 ng/mL (P < 0.05, Fig. 2
Discussion
In this study, it has been demonstrated that rat podocytes can undergo phenotypic change with increased expression of α-SMA under the influence of TGF-β1. The cell adhesion/integrin activation is needed for the induction of phenotypic change by TGF-β1.
Myofibroblasts play an important role in wound healing and scarring. Myofibroblast-like cells have been detected within glomeruli, crescents, and interstitium in experimental glomerulonephritis and humans with progressive renal diseases.3, 8, 9, 37
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