目的 探讨雌激素对宫腔粘连纤维化进程及叉头框(Fox)F2表达的影响.方法 采用0.2％Ⅰ型胶原酶消化-筛网过滤-差时贴壁分离法获取原代人子宫内膜基质细胞(HESCs)并进行鉴定.采用l0ng/ml转化生长因子β1(TGF-β1)作用于HESCs48h建立宫腔粘连细胞模型.以0、10-6、10-8、10-10、10-12mol/L雌二醇(E2)作用于模型组48h,采用qPCR及Western blotting检测α平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原蛋白(COL Ⅰ)及FoxF2的mRNA和蛋白表达.结果 免疫细胞化学法检测显示HESCs波形蛋白阳性,角蛋白18阴性.模型组α-SMA、COL Ⅰ、FoxF2的mRNA和蛋白表达均高于对照组(p＜0.05),模型构建成功.qPCR检测结果显示,与模型组相比,10-6、10-8、10-10mol/LE2组α-SMA、COL Ⅰ、FoxF2的mRNA表达除10-10mol/L E2组COL Ⅰ表达无差异外,其余各组表达均下降(P＜0.05),10-12mol/LE2组α-SMA及COL Ⅰ mRNA表达上升,但差异无统计学意义(P＞0.05),FoxF2mRNA表达下降(P＜0.05).Western blotting检测结果显示,与模型组相比,10-6、10-8、10-10mol/L E2组α-SMA、COL Ⅰ及FoxF2蛋白表达下降(P＜0.05),10-12mol/LE2组α-SMA蛋白表达上升,但差异无统计学意义(P＞0.05),COL Ⅰ及FoxF2蛋白表达下降(P＜0.05).结论 FoxF2在宫腔粘连中的表达增加.雌激素可在一定范围内逆转宫腔粘连的纤维化进程,并抑制FoxF2的表达.%Objective To investigate the effect of estrogen on the fibrosis process of intrauterine adhesions and the expression of forkhead box F2 (FoxF2).Methods Primary human endometrial stromal cells (HESCs) were obtained by separation with 0.2％ collagenase Ⅰ digestion-mesh filtration-differential adherence,and identified by immunocytochemistry.HESCs affected with 10ng/ml transforming growth factor β1 (TGF-β1) for 48 hours.HESCs in model group were affected with 0,10-6,10-8,10-10 and 10-12mol/L estrogen,the expressions of smooth muscle actin alpha (α-SMA),Collagen I (COL Ⅰ) and FoxF2 were detected by quantitative PCR (qPCR) and Western blotting.Results HESCs with high purity and good activity were obtained by using 0.2％ collagenase Ⅰ digestion-mesh filtration-differential adherence separation method.Immunocytochemistry showed positive vimentin and negative cytokeratin 18 in HESCs.The results of qPCR and Western blotting showed that the mRNA and protein expression levels of α-SMA,COL Ⅰ and FoxF2 were higher in model group than in control group (P＜0.05),the model was built successfully.qPCR revealed that the mRNA expression levels ofα-SMA,COL Ⅰ and FoxF2 were significantly lower in 10-6,10-8 and 10-10mol/L estrogen groups than in model group (P＞0.05 in 10-10mol/L estrogen group,P＜0.05 in other groups),while in 10-12mol/L estradiol group,the expression levels of FoxF2 mRNA significantly decreased (P＜0.05),and of α-SMA and COL Ⅰ mRNA increased,but no significant difference were found (P＞0.05).Compared with the model group,the protein expression levels of α-SMA,COL Ⅰ and FoxF2 in 10-6,10-8 and 10 10mol/L estrogen groups decreased,but no significant difference was found (P＜0.05),while in 10-12mol/L estradiol group,the expression levels ofα-SMA protein increased (P＞0.05),and of COL Ⅰ and FoxF2 proteins decreased (P＜0.05).Conclusions The expression of FoxF2 in intrauterine adhesions is increased.Estrogen can reverse the fibrosis process of intrauterine adhesions in a certain range and inhibit the expression of FoxF2.
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