Experimental Study On The Inhibition Role Of ActivinA On Proliferation Of Human Lens Epithelial Cells

Objective:To investigate the depressant effect of lOng/ml ACTA on generation of human lens epithelial cell line SRA01/04(HLEC SRA01/04), and the relationships with time-effect.Methods:(1)Cell culture:LDMEM medium containing 10% fetal calf serum(FBS) is used for culturing HLECs and collect exponential growth phase cells for experiment.(2)MTT colorimetry:HLECs were incubated with lOng/ml ACTA for different hours(24h,48h,72h), then calculated the cell growth inhibition rate.(3)Flow cytometric analysis (FCM):10ng/ml ACTA affect on HLECs for different hours(24h,48h,72h), then detect the earlier period cell apoptosis rate.(4) Immunofluorescence decoration:observe the morphological change of HLECs that effected on by 10ng/ml ACTA for 48h.(5)Immune fluorescence:detect caspase-3 after 10ng/ml ACTA action on HLECs for different hours(24h,48h,72h). Line statistical analysis of all results.Results:(1) The results of MTT:HLECs were treated with 10ng/ml ACTA for 24, 48,72 hours, the corresponding inhibitory ratios were 10.02%,10.91% and 18.11%. It shows that 10ng/ml ACTA can inhibit the growth of HLECs distinctly. The more longer time that HLECs were treated with 10ng/ml ACTA, the more larger inhibition ratio is. There were significant differences(P<0.05) (2) The results of FCM:HLECs were treated with 10ng/ml ACTA for 24, 48,72hours, the early apoptosis rate of HLECs were 2.3%,6.2%,9.4%, The early apoptosis rate of control group was 1.4%. The difference between the groups was statistically significant (p<0.05), the difference between the groups was the time effect relationship.(3) The results of DAPI dyeing:Morphological changes of apoptosis such as cellular shrinkage, nuclear fragmentation, nuclear dissolve would happen after affect by lOng/ml of ACTA for 48 hours. On contrary, Control group of cells had morphological rules, plump nuclei, uniform chromatin staining.(4) The results of Immune fluorescence:Caspase-3 increase gradually after HLECs were treated with 10ng/ml ACTA for 24,48,72 hours. The difference was statistically significant (p<0.05).Conclusion:10ng/ml ACTA can inhibit HLECs proliferating and induce apoptosis. That is a time-effect relationship. The ACTA is expected to be a effective drugs combating after-cataract.