| Objective 1. To explore the effects on cell proliferation of modified Zhujing prescription medicated serum in cobalt chloride induced ARPE-19 cells. 2. To explore the effects on the expression of vascular endothelium growth factor(VEGF) and hypoxia-inducible factor 1α(HIF-1α) protein of modified Zhujing prescription medicated serum in cobalt chloride induced ARPE-19 cells. To discuss the molecular mechanism of modified Zhujing prescription inhibiting the formation of choroidal neovascularization(CNV) and to provide experimental evidence to treat CNV.Methods 1. The ARPE-19 cells which cultured in vitro and logarithmic grew well were randomly assigned to the normal control group, hypoxic model group and drug intervention groups. Every drug intervention group were set up some control groups according to concentration and time. 2. To detect the effects of cytotoxicity with different time and different concentration(medicated serum group, blank serum group and Conbercept group) by CCK-8 method. 3. To establish the hypoxic cell models of cobalt chloride. 4. To detect the effects on cell proliferation of every group by CCK-8 method. 5. To observe the effects on the expression of VEGF and HIF-1α by ELISA. 6. To observe the effects on the expression of VEGF and HIF-1α protein by ELISA.Results 1. There was no toxic reaction for ARPE-19 cells with different concentration of modified Zhujing prescription medicated serum group, blank serum group and 20 μg/m L Conbercept group. Compared these groups with normal control group, there were no statistically significant differences(P>0.05). There were toxic reaction for ARPE-19 cells with greater than or equal to 40 μg/ml Conbercept groups. Compared these groups with normal control group, there were statistically significant differences(p<0.05). 2. The groups of 100 μmol/L and 200 μmol/L cobalt chloride could promote the cell proliferation. The 400μmol/L cobalt chloride group could inhibit cell proliferation. Compared these groups with normal control group, there were statistically significant differences(p < 0.05). 3. Both cell proliferation and expression of VEGF and HIF-1α of hypoxic model group were higher than normal control group. The difference between them had statistically significant(p<0.05). Except blank serum group, 5% modified Zhujing prescription medicated serum group, and 5μg/ml Conbercept group, the cell proliferation and expression of VEGF and HIF-1α of every drug intervention group were lower than hypoxic model group. The difference had statistically significant(p<0.05).Conclusions 1. Hypoxia injury can promote the cell proliferation of ARPE-19 cells and the expression of VEGF and HIF-1α. 2. Modified Zhujing prescription medicated serum can inhibit the cell proliferation and the expression of VEGF and HIF-1α of hypoxia injuried ARPE-19 cells. 3. One of the mechanism of modified Zhujing prescription treating CNV may be improve the hypoxia ischemia condition and inhibit the secretion of cell factors. |
