TGF-β Induces Matrix Metalloproteinase Secretion In Lens Epithelial Cells

Objective: (1) To determine the effect of Transforming growth factor-1(TGF-1) on the proliferation of lens epithelial cells; (2)To determine the expression of Matrix Metalloproteinases (MMPs) in lens epithelial cells; (3)To explore the effect of TGF-1 on MMPs expression in lens epithelial cells .Methods: (1) Bovine lens epithelial cells(BLEC) were primarily cultured and subcultured. MMPs activity of the cell culture media from primary culture and different passages were detected by gelatin zymography. (2) Primary culture cells were moved into plates with 96 holes (10 per hole) and cultured in 5% bovine serum culture media containing different concentration of TGF-1 (0.1, 1, 10ng/ml, final concentration). Cells cultured in 5% bovine serum without TGF-1 was set as control. 24 hours later, the cells culture media were collected and MMPs activity were detected by gelatin zymography. (3) The first passage cells were incubated in 5% bovine serum culture media containing TGF-1 (lOng/ml) for 12, 24, 36, 48, 72 hours, cells culture media were collected at indicated time. Gelatin zymography was used to detect the activity of MMPs in all cell culture media collected. Counte the residual cells. BLEC cultured in 5% bovine serum without TGF-1 was set as control. Results: (1) After TGF-1 with 0.1, 1, 10ng/ml processed for 24 hours, cell counts were respectively 1.4 0.15, 1.34 0.22, 1.3 0.17 104 and1.65 0.12xio4 in control group. Cell counts in 12, 24, 36, 48, 72 hours were respectively 1.32 0.13, 1.65 0.12, 1.75 0.16, 1.85 0.16, 2.36 0.16 xio4 in control group, and 1.04 0.1(K 129*0.17, 1.30 0.15, 1.47 022, 1.56*026 104xio4 in TGF-pl group. (2) MMPs activity were not detected in culture media from primary culture and different passages cells. (3) MMP-2 and MMP-9 activity were detected in cells media containing TGF-pl. After incubated with different concentration of TGF-(31 (0.1, 1, 10ng/ml, final concentration) for 24 hours, MMP-2 activity per 10,000 cells were respectively 699.79 60.77, 870.20 46.45> 989.50 98.44, and MMP-9 activity per 10,000 cells were respectively 171.28 17.33 ^ 233.91 25.49, 279.6 15.52. Not any MMP activity were detected in control group. (4) After TGF-(31 (lOng/ml, final concentration) were added for 12, 24, 36, 48, 72 hours, MMP-2 and MMP-9 activity increased as the time going along. MMP-2 activity were respectively 730 31.37, 990 110.0, 1102.2 46.42, 1272.8*25.08, 1639.2 39.69, and MMP-9 activity were respectively 227.2*21.65, 279.6 15.52, 346.6 36.07, 510.8 62.83, 679.4 108. Not any MMP activity were detected in control group.Conclusion: (1) TGF- 1 can restrain the proliferation of bovine lens epithelial cells. (2) MMPs activity was not expressed in normal bovine lens epithelial cells in vitro. (3) TGF- 1 can induce the expression of MMPs in BLEC, and MMPs activity was dependent on time and doseconcentration.