| Object To investigate the effects of bilberry extract on the expression of MMP-2 and Collagen I protein in human fetal scleral fibroblast?HFSF? in vitro.Methods The frozen HFSF cells were subcultured, and inverted microscope was used to observe their biological properties. After HFSF cells were treated with bilberry extract at concentrations of 0, 10-6, 10-5, 10-4, 10-3, 10-2, 10-1, 1mg/m L respectively for 6h, 8h, 12 h, 24 h,48h, the effects of bilberry extract on the proliferation of the cells were tested by MTT assay.After HFSF cells were treated with bilberry extract at concentrations of 0, 10-1mg/m L respectively for 12 h, the effects of bilberry extract on the cell cycle and apoptosis were detected by flow cytometry with PI staining and Annexin V-FITC/PI, respectively. The m RNA levels of MMP2, COL1A1, COL1A2 gene expression were determined by quantitative reverse transcription PCR?q RT-PCR?. The protein levels of matrix metalloproteinase 2?MMP-2? and Collagen I expression were detected by western blot.Results Mature cultured HFSF cells tended to be in fusiform, spiral or beam-shaped patterns. The results of MTT assay showed that after treated with bilberry extract, the proliferation of HFSF cells was highest with a concentration of 10-1mg/m L at 12 h. The results of flow cytometry showed bilberry extract increased the number of HFSF cells in G2 stage,and it did not affect cell apoptosis. The results of q RT-PCR showed that bilberry extract down-regulated the expression of MMP2 gene. The results of Western Blot showed that bilberry extract inhibited the expression of MMP-2 protein, but the expression of Collagen I protein was not detected. Additionally, after HFSF cells were treated with TGF-?2, the results of Western Blot showed that the expression of Collagen I protein was detected.Conclusion Bilberry extract promotes the growth of cultured HFSF cells in vitro and the expression of MMP-2 protein was inhibited. TGF-?2 induced the expression of Collagen I protein in HFSF cells. |
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