The Inhibitory Effect Of The 8-chloroadenonosine On Proliferation Of Human Lens Epithelial Cells In Vitro

【Objective】To observe the inhibiting effect of 8-chloroadenonosine on human lens epithelial cell growth. And to search for a new drug to inhibit posterior capsular opacification after cataract surgery.【Materials and Methods】The second or third passage of human lens epithelial cells growth were used to be treated with different concentrations of 8-chloroadenonosine (2, 4, 8, 16μmol/L). 1. MTT assay: The human lens epithelial cells were cultured in four 96 well plates for 24, 48, 72, 96 hours. There were 6 groups in every 96 well plate, and every group had 4 repeat holes. The first and the second group were the vacant control group and the negative control group respectively. The first group only had culture media. The second group had cells and culture media. The last 4 test groups had not only cells and culture media but also 8-chloroadenonosine with different concentrations (2,4,8,16μmol/L). The effect of 8-chloroadenonosine on human lens epithelial cells growth were determined by MTT assay 24h, 48h, 72h and 96h after treatment. 2. Flow cytometer: The cell cycle of negative control group and test groups (the concentration was 2,4,8,16μmol/L respective) were determined by Flow cytometer (FCM) 48h after treatment.【Results】1. MTT assay: Figure out the inhibitive rate of 8-chloroadenonosine to human lens epithelial cell in every test group according to the value of optical density (OD) data. By doing S-N-K analysis, we can find that only when 8-chloroadenonosine on 4~8μmol/L concentrations and the test time were 48~72h, the inhibitive rates had such significant difference: the significant difference between two groups with neighbour concentrations in the same test time; and the significant difference between two groups with neighbour test time in the same concentrations. So 8-chloroadenonosine can inhibit the growth of human lens epithelial cell with the concentrations from 4μmol/L to 8μmol/L and the test time from 48h to 72h. And the effect depends upon the time and concentrations. 2. Flow cytometer: The cell cycle of negative control group and test groups were determined by Flow cytometer (FCM) 48h after treatment. By doingχ2 analysis we can find that the cell rate of G1 term had significant difference between negative control group and those test groups whose concentrations were amount to or superior to 4μmol/L. And there were significant difference between 4μmol/L group and 8μmol/L but no significant difference between 8μmol/L group and 16μmol/L. This result coincided with the MTT assay.【Conclusion】8-chloroadenonosine can inhibit the growth of human lens epithelial cell at the concentration from 4μmol/L to 8μmol/L after treating 48~72h. And the effect depends upon the times and concentration. This indicated that 8-chloroadenonosine are perhaps helpful in treating posterior capsular opacification after cataract surgery.