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Niu ZW, Zhao HW: [The effects of transforming growth factor-beta1 on the differentiation of BALB/c 3T3 fibroblasts into myofibroblasts and the change of Egr-1 expression]. Hepatology. 2010 Mar;51(3):986-95. OBJECTIVE: To investigate the effects of transforming growth factor (TGF)-beta (1) on fibroblasts differentiation into myofibroblasts, and the change of Egr-1 expression in the course of differentiation in BALB/c 3T3 fibroblasts. METHODS: BALB/c 3T3 fibroblasts of the experiment group were treated with 10 ng/ml TGF-beta (1) and those of the control group were treated without TGF-beta (1). The expression of alpha-smooth muscle actin (SMA) was detected by flow cytometer after incubation for 24, 48, and 72 h. The expression of Egr-1 was detected by immunocytochemistry and Western blot method after 10 ng/ml TGF-beta (1) incubation for 15, 30, 60, 90, 120, 180, and 240 min. RESULTS: The percentage of alpha-SMA positive cells was (6.65 +/- 0.48)% when cells were treated with 10 ng/ml TGF-beta (1) for 24 h, and the control group was (5.53 +/- 0.62)%, which showed no statistical significance. However the percentage was up to (28.38 +/- 3.60)% after treated with TGF-beta (1) for 48 h, and the control group was (9.49 +/- 0.21)%; At 72 h, the percentage was (36.04 +/- 0.73)% and the control group was (15.23 +/- 0.33)%; The difference was statistically significant. Immunocytochemistry showed that Egr-1 protein positive signal was brown in color. The color was light in the control group. The gray-level of the control group was 38 +/- 18 measured by Western blot when cells were treated without TGF-beta (1). After treated with 10 ng/ml TGF-beta (1) for 15 min, the level was up to 55 +/- 26, which had no statistical significance. The level was increased with longer incubation. It was 115 +/- 14 at 60 min and reached its peak level 129 +/- 22 at 90 min, which was significantly higher than the control group. The level was decreased after 90 min and reached neap 39 +/- 7 at 240 min. CONCLUSION: TGF-beta (1) increased the expression of alpha-SMA in BALB/c 3T3 fibroblasts and affected the Egr-1 protein expression during the early course of fibroblasts differentiation into myofibroblasts. |
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