Effects Of Estrogen On The Fibrosis Process Of Intrauterine Adhesions And The Expression Of Forkhead Box F2

Background:Intrauterine adhesions refers to the partial or complete adhesions in the uterine cavity because of the endometrial basal layer injury due to trauma,infection or other causes.Recent studies have shown that endometrial fibrosis is the main pathological changes of intrauterine adhesions.In other words,intrauterine adhesions is an endometrial fibrosis disease in essence.Forkhead box F2(FoxF2)is an important transcriptional regulator in process of organ development,extracellular matrix synthesis and epithelial-mesenchymal cell transformation,involve in the development of multiple organ fibrosis diseases.However,the role of FoxF2 in the process of intrauterine adhesions has not been reported in releated literature.Estrogen is the most important sex hormone present in women.In the treatment of intrauterine adhesions,estrogen always has been as an empirical treatment widely used by domestic and foreign clinicians.However,the effects of estrogen are different and the related basic research is still absent.There are still a lot of controversies about estrogen’s application dose.The purpose of this research was to investigate the expression of FoxF2 in intrauterine adhesions and further explore the effects of estrogen on fibrosis process of intrauterine adhesions and the expression of FoxF2.Chapter 1 Expression and significance of forkhead box F2 in the cell model of intrauterine adhesionsObjective:To investigate the expression of FoxF2 in the cell model of intrauterine adhesions.Methods:1.Primary cultured human endometrial stromal cells(HESCs).2.Immunocytochemical staining was performed to identified the cells.3.Establish a cell model of intrauterine adhesions.Applied 0 and 10 ng/ml transforming growth factor β1(TGF-β1)to HESCs for 48 hours as control group and model group.4.Real-time quantitative PCR and western blotting was performed to detect the mRNA and protein expression of α-smooth muscle actin(a-SMA),collagen I(COLI)and FoxF2.Results:1.We can obtain HESCs with stable morphology and great growth activity by primary culture.2.Immunocytochemical staining showed that cells are positive for vimentin and negative for cytokeratin 18.The positive cell rate of vimentin was high.It proved to be HESCs with high purity.3.Compared with the control group,the mRNA and protein expression of a-SMA,COLI and FoxF2 in model group were significantly increased,and the difference was statistically significant.Conclusion:1.10 ng/ml TGF-β1 used in HESCs can induce myofibroblast-like changes and increase the expression of fibrosis markers a-SMA and COLI,so the cell model of intrauterine adhesions was established.2.FoxF2 is highly expressed in intrauterine adhesions,which suggesting that FoxF2 may participate in the fibrosis process of intrauterine adhesions.Chapter 2 Effects of estrogen on the fibrosis process of intrauterine adhesions and the expression of forkhead box F2Objective:To investigate the effects of estrogen in different concentrations on the fibrosis process of intrauterine adhesions and the expression of FoxF2.Methods:1.Configure working solution of estrogen in 10-6,10-8,10-10 and 10-12 mol/L.2.This experiment was divided into five groups:model group,10-6 mol/L E2 group,10’8 mol/L E2 group,10’10 mol/L E2 group and 10-12 mol/L E2 group.Firstly,we have to apply 10 ng/ml TGF-β1 to HESCs for 48 hours in order to establish the cell model of intrauterine adhesions.And then,we applied 0,10-6,10-8,10-10 and 10-12 mol/L estrogen on cell model of intrauterine adhesions in order to investigate the effects of estrogen in different concentrations on the fibrosis process of intrauterine adhesions and the expression of FoxF2.3.Real-time quantitative PCR and western blotting was performed to detect the mRNA and protein expression of α-SMA,COLI and FoxF2.Results:1.Real-time quantitative PCR results:compared with the model group,the mRNA expression of a-SMA,COLI and FoxF2 in 10-6,10-8,10-10 mol/L E2 group decreased consistently(P<0.05 in each group of each mRNA,except for COLI in 10’10 mol/L E2 group).While in 10-12 mol/L E2 group,the mRNA expression of a-SMA and COLI increased(P>0.05),the mRNA expression of FoxF2 decreased(P<0.05).2.Western blotting results:compared with the model group,the protein expression of a-SMA,COLI and FoxF2 in 10-6,10-8,10-10 mol/L E2 group decreased consistently(P<0.05).While in 10’12 mol/L E2 group,the protein expression of a-SMA increased(P>0.05),the protein expression of COLI and FoxF2 decreased(P<0.05).Conclusion:17β-estradiol can down-regulate the expression of α-SMA and COLI in a certain extent,reverse the fibrosis process of intrauterine adhesions and inhibit the expression of FoxF2.