| Objective To investigate cell localization,protein expression and its possible association of ERα、ERβ、AP-1、VEGF in ectopic endomeitrium and eutopic endomeitrium of women with endometriosis(EMT).To compare these proteins expression in different menstrual stages and explore the possible pathogenesis of EMT,to provide theoretical foundation for clinical targeted treatment of EMT.Methods(1)40 patients with ovarian endometriosis undergoing laparoscopic cystectomy allocated into the EMT experimental group,proliferative phase(n=16),secretory phase(n=24),ectopic endometrium tissue is group A and eutopic endometrium tissue is group B.37 patients with infertility due to fallopian tube were selected at the same time,proliferative phase(n=22),secretory phase(n=15),normal endometrium tissue removal by hysteroscope were the control group.(2)12 patients in the experimental group were selected,proliferative phase(n=6),secretory phase(n=6).12 in the control group,proliferative phase(n=6),secretory phase(n=6).ERα,ERβ,AP-1,and VEGF were quantitatively detected in the ectopic endomeitrium 、 eutopic endomeitrium and normal endometrium by Western blot.Compare protein expression differences between proliferative and secretory phases of ERα,ERβ,AP-1,and VEGF in the ectopic endomeitrium、eutopic endomeitrium and normal endometrium.Results1.The results of immunohistochemistry results1.1 ERα was expressed in the ectopic endometrium,eutopic endometrium and normal endometrium.The positive rates were 48.0%,55.8%,and 37.3%,and both glandular epithelial cells and stromal cells were localized.The expression of ERα in the eutopic endometrium was higher in the ectopic endometrium and the difference was statistically significant(χ~2=4.812,P=0.028<0.05).Comparison between experimental group and control group,the expression of ERα in the ectopic endometrium was higher in the normal endometrium and the difference was statistically significant(χ~2=8.990,P=0.003<0.05).The expression of ERα in the eutopic endometrium was higher in the normal endometrium and the difference was statistically significant(χ~2=26.281,P<0.001).1.2 ERβ was expressed in the ectopic endometrium,eutopic endometrium and normal endometrium.The positive rates were 55.0%,50.2%,33.2%,and both glandular epithelial cells and stromal cells were localized.The expression of ERβ in the ectopic endometrium was not statistically different from that of the eutopic endometrium(χ2=1.801,P=0.179).Comparison between experimental group and control group,the expression of ERβ in the ectopic endometrium was higher in the normal endometrium and the difference was statistically significant(χ2=36.831,P<0.001).The expression of ERβ in the eutopic endometrium was higher in the normal endometrium and the difference was statistically significant(χ2=22.809,P<0.001).1.3 c-Jun was expressed in the ectopic endometrium,eutopic endometrium and normal endometrium.The positive rates were 88.5%,86.8%,79.5%,and both glandular epithelial cells and stromal cells were localized.The expression of c-Jun in the ectopic endometrium was not statistically different from that of the eutopic endometrium(χ2=0.565,P=0.452).Comparison between experimental group and control group,the expression of c-Jun in the ectopic endometrium was higher in the normal endometrium and the difference was statistically significant(χ2=11.782,P=0.001<0.05).The expression of c-Junin the eutopic endometrium was higher in the normal endometrium and the difference was statistically significant(χ2=7.325,P=0.007<0.05)。1.4 VEGF was expressed in the ectopic endometrium,eutopic endometrium and normal endometrium.The positive rates were 83.2%,81.2%,64.9%,and both glandular epithelial cells and stromal cells were localized.The expression of VEGF in the ectopic endometrium was not statistically different from that of the eutopic endometrium(χ2=0.548,P=0.459>0.05).Comparison between experimental group and control group,the expression of VEGF in the ectopic endometrium was higher in the normal endometrium and the difference was statistically significant(χ2=34.124,P<0.001).The expression of VEGF in the eutopic endometrium was higher in the normal endometrium and the difference was statistically significant(χ2=26.415,P<0.001).2.The results of Western blot2.1 The ERα IOD values of ectopic endometrium and eutopic endometrium were(593.92±8.14)and(623.46±13.19),respectively,higher than those of normal endometrium(455.13±18.86),and the differences were statistically significant(t=33.098,P<0.05;t=35.829,P<0.05);ERα IOD values of eutopic endometrium were higher than that of ectopic endometrium,and the difference was statistically significant(t=9.337,P<0.05).The intensity of ERα expression in normal intima was periodic,and the proliferative phase(469.92±11.29)was higher than the secretory phase(440.33±11.80).The difference was statistically significant(t=6.277,P<0.05).However,the experimental group had no such periodical changes: Ectopic endometrium(proliferative phase: 595.17±7.64,secretory phase: 592.67±8.78,t=0.745,P=0.464),eutopic endometrium(proliferative phase: 625.17±12.86,secretory phase: 621.75±13.86,t=0.626,P=0.538).2.2 The ERβ IOD values of ectopic endometrium and eutopic endometrium were(645.21±8.86)and(642.83±6.86),respectively,which were higher than those of normal endometrium of the control group(438.21±31.25).The difference was statistically significant(t=38.134,P<0.05;t=38.567,P<0.05);There was no significant difference in ERβ IOD between eutopic endometrium and eutopic endometrium(t=1.038,P=0.305).The expression of ERβ in the normal endometrium of the control group was characterized by a periodical change.The proliferative phase(460.17±10.07)was higher than the secretory phase(417.58±14.64).The difference was statistically significant(t=8.300,P<0.05).However,the experimental group had no such periodical changes: Ectopic endometrium(proliferative phase: 646.08±10.58,secretory phase:644.33±7.11,t=0.476,P=0.639),eutopic endometrium(proliferative phase: 643.08±7.49,secretory phase: 642.58±6.50,t=0.175,P=0.863).2.3 The ERβ/ERα ratios of ectopic endometrium and eutopic endometrium IOD values were(1.087±0.231)(1.032±0.258),respectively,which were significantly higher than those of the control normal endometrium(0.964±0.349).The difference was statistically significant(t =14.347,P<0.05;t=7.606,P<0.05);ERβ/ERα ratio(1.087±0.231)in endometriosis IOD was higher than that in eutopic endometrium(1.032±0.258).The difference was statistically significant.(t=7.786,P<0.05).2.4 The c-Jun IOD values of ectopic endometrium and eutopic endometrium were(622.29±9.02)and(618.70±9.79),respectively,which were higher than the normal endometrium IOD values(428.58±14.86)in the control group.The differences were statistically significant.Significance(t=28.22,P<0.05;t=51.56,P<0.05);There was no significant difference in c-Jun IOD between eutopic endometrium and eutopic endometrium(t=1.310,P=0.197).The expression of c-Jun in the normal intima of the control group was characterized by a periodic variation,with the proliferative phase(439.25±12.05)being higher than the secretory phase(417.92±8.25).The difference was statistically significant(t=53.59,P<0.05).However,the experimental group had no such periodical changes: Ectopic endometrium(proliferative phase: 625.00±10.41,secretory phase: 619.58±6.78,t=1.511,P=0.145),eutopic endometrium(proliferative phase: 621.83±9.01,secretory phase: 616.92±10.97,t=1.199,P=0.243).2.5 The ERβ/ERα values of the ectopic intimal IOD values were negatively correlated with the c-Jun IOD values(r=-0.555,P<0.05),whereas the ERβ/ERα values and c values of the intimal IOD values were positive.The correlation of Jun IOD values(r=-0.008),the correlation between normal endometrium IOD values of ERβ/ERα and c-Jun IOD values(r=0.233)were not statistically significant(P=0.971,P =0.274).2.6 The VEGF IOD values of ectopic endometrium and eutopic endometrium were(613.21±9.62)and(612.14±8.50),respectively,which were higher than those of normal endometrium in the control group(430.67±22.39).The differences were statistically significant(t=36.70,P<0.05;t=37.12,P<0.05).There was no significant difference in VEGF IOD between ectopic and eutopic endometrium in the group(t=0.413,P=0.618).The intensity of VEGF expression in the normal endometrium of the control group was periodically changed.The proliferative phase(449.92±10.87)was higher than the secretory phase(411.42±11.02).The difference was statistically significant(t=8.616,P<0.05).However,the experimental group had no such periodical changes: Ectopic endometrium(proliferative phase: 615.75±8.80,secretory phase: 610.67±10.10,t=1.314,P=0.202),eutopic endometrium(proliferative phase: 613.75±9.23,secretory phase: 610.50±7.75,t=0.934,P=0.360).2.7 Correlations between ERβ/ERα values and VEGF IOD values of IOD values were not statistically significant in ectopic endometrium(r=-0.225),eutopic endometrium(r=0.127),and normal endometrium(r=0.327).Significance(P<0.292,P<0.553,P<0.119).Conclusion1.In patients with EMT,the protein expression of ERα in eutopic endometrium is highly.In ectopic endometrium,the effect of ERβ is higher than that of ERα.2.In ectopic and eutopic endometrium of patients with EMT,AP-1 and VEGF are highly expressed,and the ratio of ERβ and ERα is negatively correlated with AP-1,AP-1 and VEGF play an important role in the development of EMT.3.The intensity of the expression of ERα,ERβ,AP-1,and VEGF in normal endometrium showed periodic characteristics,and the proliferative phase is higher than the secretory phase.However,the intensity of expression of these proteins in the ectopic endometrium and eutopic endometrium of EMT do not have this periodicity.Changes suggest that the biological behavior of EMT endometrial cells has changed. |
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