Inhibitive effect of Decorin on proliferation of rabbit lens epithelial cells
Recent Advances in Ophthalmology(2011)
Abstract
Objective To study the inhibitive effect of Decorin on proliferation of rabbit lens epithelial cells(LEC) and its dose-dependent,time-dependent manner,and offer new options for preventing and treating of posterior capsular opacification. Methods The growth of the LEC after treating with different concentrations(0.1 mg·L-1,1.0 mg·L-1,10.0 mg·L-1) of Decorin were examined by MTT method at different time points,and the growth cycle was checked with flow cytometry,ELISA was used to measure the levels of TGF-β in culture medium supernatant,RT-PCR was applied to identity the expression of TGF-β mRNA,and immunocytochemical methods was used to observe the α-smooth muscle actin(α-SMA). Results ELISA showed the level of TGF-β in per million cells in control group,0.1 mg·L-1,1.0 mg·L-1 and 10.0 mg·mL-1 Decorin group were(427.24±41.34)ng,(236.01±28.63)ng,(117.86±18.14)ng,(111.72±26.72)ng,the level of TGF-β of the Decorin groups were significantly lower than that of the control group(all P0.05).MTT assay results showed the inhibitive ratio of control groups,0.1 mg·L-1,1.0 mg·L-1 and 10.0 mg·L-1 Decorin group at 24 hours were (3.62±1.34)%,(4.14±2.08)%,(12.93±2.47)%,(23.47±1.78)%,there were statistical differences(all P0.05),and the inhibitive ratios were increased with the time prolonged of Decorin treating,there were statistical differences(all P0.05).With the drug concentration increased and treating time prolonged,LEC apparent G0/G1 phase blocked,the percentage of cells in G0/G1 phase was significantly increased(P0.05).RT-PCR showed the expression of TGF-β mRNA in Decorin group was significantly lower than that of the control group at the same time,which was similar to that detected by MTT.Immunocytochemical method showed the expression of α-SMA in control group was higher than that in Decorin groups,cells were hypertrophy and cytoplasm were brown.The Decorin showed significant inhibition. Conclusions Decorin can effectively inhibit the growth of LEC and induce their apoptosis in dose-dependent and time-dependent manner,which is expected to become the drug for prevention and treatment of posterior capsular opacification.
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Key words
decorin,rabbit lens,epithelial cells
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