The Effect Of EGCG On Proliferation And Apoptosis Of Pterygium Fibroblasts

Objective:Through the primary culture of pterygium fibroblasts(HPFS),the effects of EGCG(-)epigalocatechin gallate,a green tea extract,on the proliferation and apoptosis of HPFS were observed.The effects of different doses and time of EGCG on the proliferation of HPFS were analyzed.The feasibility of EGCG in the treatment of pterygium was discussed,which provided a new direction for the adjuvant treatment of pterygium.Methods:1.The primary culture of pterygium fibroblasts(HPFS)was carried out by the method of tissue block adherent culture;2.Identification of purified HPFS cells with vimentin and CK by cellular immunohistochemistry;3.CCK-8 method was used to detect the inhibitory effect of different concentrations of EGCG and the same concentration of EGCG at different time on the proliferation of pterygium fibroblasts;4.The effect of different concentrations of EGCG on the apoptosis of pterygium fibroblasts was detected by flow cytometry(FCM);5.SPSS 22.0 was used for statistical analysis:the data obtained was expressed as X±S,the data of each group was analyzed by repeated analysis of variance,and the comparison between each experimental group and the control group was by t test.p<0.05 is the criterion for statistical significance.Results:1.The primary culture of HPFs cells was observed under an inverted microscope:after about 5 days of culture,the cells crawled out from the periphery of the tissue block,and the cells were long spindle-shaped and scattered around the tissue block.2.Identification of pterygium fibroblasts(cell immunohistochemistry):Cells were subcultured to 3?5 generations for identification,stained cells were observed under a light microscope.Most cells were long spindle under an inverted microscope and a few were flat,a large number of Brown yellow particles can be seen in the cytoplasm(Vimentin staining is positive).3.The morphology of HPFs changed significantly under the effect of different concentrations of EGCG EGCG can inhibit the proliferation of HPFs cells in a dose-and time-dependent manner.When the concentration of EGCG reaches 500?M for 72 hours,the inhibition rate of HPFs cells can reach 93.39%.4.Flow cytometry showed that EGCG(100?M,and 200?M)acted on HPFs for 48 hours,the apoptosis rates were:3%,20%?Conclusions:1.EGCG can inhibit the proliferation of HPFs in a time-and concentration-dependent manner.2.EGCG can induce apoptosis of HPFs in a dose-dependent manner for 48 hours.3.EGCG can inhibit the growth of pterygium by inhibiting the proliferation of HPFs and inducing its apoptosis pathway.