Effects Of 17?-estradiol On The Expression Of TGFBI And Periostin And Their Interaction

Purpose:In a pedigree of Avellino corneal dystrophy,we found that the visual impairment of female patients was more severe than that of male patients.This research assessed the effects of 17?estradiol on the expression of Periostin and TGFBI,and analyzed their interaction in immortalized corneal epithelial cells,to investigate the role and molecular mechanism of TGFBI and Periostin in Avellino corneal dystrophy.Methods:The spontaneously derived human corneal epithelial cell line1?SDHCEC1?were treated with different concentrations of estrogen(10^-7?10^-8?10^-9?10^-10 mol/L)for a period of time,total proteins were extracted for Western blot,and the total RNA was extracted for real-time fluorescence quantitative PCR?q RT-PCR?,and then protein level and mRNA level of TGFBI and Periostin were analyzed.SDHCEC1 were grown on coverslips,and subjected to immunofluorescence staining.Fluorescence intensity and colocalization of TGFBI protein and Periostin protein were observed under a laser scanning confocal microscope after estrogen treatment.Finally,the interaction of TGFBI protein and Periostin protein in SDHCEC1 was verified by co-immunoprecipitation assay.Result:1.The results of Western blot and qRT-PCR showed that the expression of TGFBI and Periostin were dose dependent,and increased after treatment with different concentrations(10^-7?10^-8?10^-9?10^-10 mol/L)of 17?estradiol?P<0.05?.2.Immunofluorescence results:The average optical density of TGFBI and Periostin in 17?estradiol-treated groups(10^-7 mol/L and10^-8mol/L)were significantly higher than control group?P<0.05?,and showed that they were well co-located in the cytoplasm of SDHCEC1.4.Co-immunoprecipitation suggested that TGFBI and Periostin have interaction.Conclusion:1.17?estradiol promotes the expression of TGFBI and Periostin protein dose-dependently in SDHCEC1.2.TGFBI and Periostin have interaction in SDHCEC1.